No full-text available
To read the full-text of this research,
you can request a copy directly from the authors.
Hydrogen-rich water prevents progression of nonalcoholic steatohepatitis and accompanying hepatocarcinogenesis in mice
Abstract
Unlabelled:
Oxidative stress is a strong contributor to the progression from simple fatty liver to nonalcoholic steatohepatitis (NASH). Molecular hydrogen is an effective antioxidant that reduces cytotoxic reactive oxygen species. In this study, we investigated the effects of hydrogen-rich water and the drug pioglitazone on the progression of NASH in mouse models. A methionine-choline-deficient (MCD) diet mouse model was prepared. Mice were divided into three experimental groups and fed for 8 weeks as follows: (1) MCD diet + control water (CW group); (2) MCD diet + hydrogen-rich water (HW group); and (3) MCD diet mixed with pioglitazone (PGZ group). Plasma alanine aminotransferase levels, hepatic expression of tumor necrosis factor-α, interleukin-6, fatty acid synthesis-related genes, oxidative stress biomarker 8-hydroxydeoxyguanosine (8-OHdG), and apoptosis marker terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL)-positive cells in the liver were decreased in the HW and PGZ groups. The HW group showed a smaller decrease in hepatic cholesterol; however, stronger antioxidative effects in serum and lower peroxisome proliferator-activated receptor-α expression in the liver were seen in comparison with the PGZ group. We then investigated the effects of hydrogen in the prevention of hepatocarcinogenesis in STAM mice, known as the NASH-related hepatocarcinogenesis model. Eight-week-old male STAM mice were divided into three experimental groups as follows: (1) control water (CW-STAM); (2) hydrogen-rich water (HW-STAM); and (3) pioglitazone (PGZ-STAM). After 8 weeks, hepatic tumors were evaluated. The number of tumors was significantly lower in the HW-STAM and PGZ-STAM groups than in the CW-STAM group. The maximum tumor size was smaller in the HW-STAM group than in the other groups.
Conclusion:
Consumption of hydrogen-rich water may be an effective treatment for NASH by reducing hepatic oxidative stress, apoptosis, inflammation, and hepatocarcinogenesis.
... The former can be categorized into investigations using cultured cancer cells, transplants of animal-specific tumor lines (allogeneic transplants), xenografts of human tumor lines in immunocompromised animals, or tumors induced in animals via exposure to UV or ionizing irradiation [25][26][27][28][29][30][31][32][33][34][35][36]. There are also reports of animals fed with a high-fat diet to induce nonalcoholic steatohepatitis (NASH) and evaluate its efficacy in the progression of liver cancer [37]. In addition, since the growth of cancer is accompanied by angiogenesis, the efficacy of cultured cells on angiogenesis has also been reported [38]. ...
... [36] High-fat diet-induced liver cancer H2-rich water showed carcinogenic effects in an experimental system that progressed from NASH to fibrosis and liver cancer. [37] Angiogenesis Co-culture experiment of 2 cell lines Electrolyzed reduced H2 water inhibited lumen formation via suppression of VEGF expression in cultured cells. Inhalation of H2 gas prolonged progression-free survival and overall survival in patients with stage IV, and the mechanism involved the recovery of CD8 + T cells. ...
... Oxidative stress has been strongly implicated in the pathogenesis from simple fatty livers to NASH, fibrosis, and hepatocarcinoma. Kawai et al. investigated the inhibitory effect of H2 on the progression to hepatocarcinogenesis in a STAM mouse model [37]. Two-day-old mice were treated with a single dose of streptozotocin to reduce insulin secretion and fed a high-fat diet from 4 weeks of age. ...
While many antitumor drugs have yielded unsatisfactory therapeutic results, drugs are one of the most prevalent therapeutic measures for the treatment of cancer. The development of cancer largely results from mutations in nuclear DNA, as well as from those in mitochondrial DNA (mtDNA). Molecular hydrogen (H2), an inert molecule, can scavenge hydroxyl radicals (·OH), which are known to be the strongest oxidizing reactive oxygen species (ROS) in the body that causes these DNA mutations. It has been reported that H2 has no side effects, unlike conventional antitumor drugs, and that it is effective against many diseases caused by oxidative stress and chronic inflammation. Recently, there has been an increasing number of papers on the efficacy of H2 against cancer and its effects in mitigating the side effects of cancer treatment. In this review, we demonstrate the efficacy and safety of H2 as a novel antitumor agent and show that its mechanisms may not only involve the direct scavenging of ·OH, but also other indirect biological defense mechanisms via the regulation of gene expression.
... Molecules that are elevated by H2 administration[42]. ...
... Molecules that are elevated by H 2 administration[42]. ...
Stem cell therapy has emerged as a promising avenue for regenerative medicine, offering the potential to treat a wide range of debilitating diseases and injuries. Among the various types of stem cells, mesenchymal stem cells (MSCs) have garnered significant attention due to their unique properties and therapeutic potential. In recent years, researchers have been exploring novel approaches to enhance the effectiveness of MSC-based therapies. One such approach that has gained traction is the priming of MSCs with molecular hydrogen (H2). This article delves into the fascinating world of mesenchymal stem cell priming with molecular hydrogen and the potential benefits it holds for regenerative medicine.
... Due to selective antioxidants, hydrogen has a significant inhibitory effect on tumor control [12]. Animal models of hepatocellular carcinoma (HCC) have shown that hydrogen molecules can significantly reduce the incidence of hepatocellular carcinoma, decrease tumor volume, and reduce oxidative stress [13]. Hydrogen-enriched water significantly inhibits renal carcinoma induced by ferric nitrilotriacetate in Wistar rats [14]. ...
... Hydrogen-rich water significantly inhibited inflammatory effects, vascular endothelial growth factor (VEGF) expression, and signaling transcriptional activator in rat kidney 3-phosphorylation. It significantly inhibited the expression of proliferating cell nuclear antigens [13]. Frajese found that hydrogen-enriched electrolyzed water induced apoptosis in breast cancer cells and inhibited the expression of tyrosine kinase receptor 2 (ErbB2/neu), indicating that hydrogen-enriched electrolyzed water did not significantly affect the extracellular regulation of protein kinases (ERK1/2) and protein kinase B (AKT) [15]. ...
Cancer is still one of the major factors threatening public health, with morbidity and mortality rates at the forefront of the world. Clinical drawbacks, such as high toxicity and side effects of drug therapy, and easy recurrence after surgery affect its therapeutic effect. Gas signaling molecules are essential in maintaining biological homeostasis and physiological functions as specific chemical substances for biological information transfer. In recent years, the physiological regulatory functions of gas molecules in the cancer process have been gradually revealed and have shown broad application prospects in tumor therapy. In this paper, standard gas therapies are classified and introduced. Taking H2, CO2, NO, CO, H2S, and SO2 gases as examples, the research progress and application of gas therapies in malignant tumors are mainly introduced in terms of biological characteristics, anticancer mechanisms, and treatment strategies. Finally, the problems and prospects for developing gases as anticancer drugs are outlined.
... Some of these studies investigated the dose effect of H 2 and showed that higher doses of H 2 administration demonstrated better therapeutic effects in most cases [60,62], even though a low dose of H 2 showed better effects for some indices (e.g., plasma ALT and AST levels) [60]. Moreover, the study by Li et al. demonstrated that a longer duration of H 2 intervention exhibited more obvious improvements [61]. Consistently, prolonged hydrogen release from a nanocapsule exerted more profound effects [62]. ...
... HO-1 is an important mediator of the cross-talk between anti-inflammatory and anti-oxidative pathways. It has been reported that H 2 could inhibit the lipopolysaccharide (LPS)-induced production of TNF-α and other inflammatory cytokines via an HO-1/interleukin 10 (IL-10)-independent pathway [61]. As an inducible isoform of heme oxygenases, HO-1 catalyzes the oxidative conversion of heme to carbon monoxide (CO), ferrous ions, and biliverdin (BV), which is reduced to bilirubin-IXα (BR) by biliverdin reductase (BVR) [86]. ...
Oxidative stress and chronic inflammation have been implicated in the pathophysiology of metabolic diseases, including diabetes mellitus (DM), metabolic syndrome (MS), fatty liver (FL), atherosclerosis (AS), and obesity. Molecular hydrogen (H2) has long been considered a physiologically inert gas. In the last two decades, accumulating evidence from pre-clinical and clinical studies has indicated that H2 may act as an antioxidant to exert therapeutic and preventive effects on various disorders, including metabolic diseases. However, the mechanisms underlying the action of H2 remain unclear. The purpose of this review was to (1) provide an overview of the current research on the potential effects of H2 on metabolic diseases; (2) discuss the possible mechanisms underlying these effects, including the canonical anti-oxidative, anti-inflammatory, and anti-apoptotic effects, as well as suppression of ER stress, activation of autophagy, improvement of mitochondrial function, regulation of gut microbiota, and other possible mechanisms. The potential target molecules of H2 will also be discussed. With more high-quality clinical trials and in-depth mechanism research, it is believed that H2 will eventually be applied to clinical practice in the future, to benefit more patients with metabolic disease.
... Thus, we wonder whether earlylife antibiotics exposure also blunts LrNK-mediated anti-tumor effects. To address this, we introduced both c-myc/AKT-driven 37,38 (Fig. 2a) and STZ-HFD-induced 39 (Supplementary Fig. 5a) hepatocellular carcinoma (HCC) mouse model. It was found that early-Abx mice developed more severe liver tumors than control mice in both HCC models (Fig. 2b, c, Supplementary Fig. 5b, c). ...
... For STZ-HFD induced HCC model, a mouse model mimicking nonalcoholic fatty liver disease -related HCC 39 , control or early-Abx male mice was induced by a single subcutaneous injection of 200 μg Streptozocin (Sigma) at 2 days after birth and feeding with HFD (cat#: D12492, Rodent diet with 60 kcal% fat) after 4 weeks of age. At month 5, the mice were sacrificed and tumor nodes in murine livers were photographed, and the phenotype and function of LrNK cells were analyzed. ...
Liver-resident natural killer cells, a unique lymphocyte subset in liver, develop locally and play multifaceted immunological roles. However, the mechanisms for the maintenance of liver-resident natural killer cell homeostasis remain unclear. Here we show that early-life antibiotic treatment blunt functional maturation of liver-resident natural killer cells even at adulthood, which is dependent on the durative microbiota dysbiosis. Mechanistically, early-life antibiotic treatment significantly decreases butyrate level in liver, and subsequently led to defective liver-resident natural killer cell maturation in a cell-extrinsic manner. Specifically, loss of butyrate impairs IL-18 production in Kupffer cells and hepatocytes through acting on the receptor GPR109A. Disrupted IL-18/IL-18R signaling in turn suppresses the mitochondrial activity and the functional maturation of liver-resident natural killer cells. Strikingly, dietary supplementation of experimentally or clinically used Clostridium butyricum restores the impaired liver-resident natural killer cell maturation and function induced by early-life antibiotic treatment. Our findings collectively unmask a regulatory network of gut-liver axis, highlighting the importance of the early-life microbiota in the development of tissue-resident immune cells.
... H 2 administration is increasingly being explored as part of anticancer treatments in animals and humans due to its remarkable capacity to scavenge free radicals. Drinking HW reduced oxidative stress, inflammation, and apoptosis in rats, preventing the development of non-alcoholic steatohepatitis and its associated carcinogenesis [43]. Treatment of cancer by anti-tumour drugs and radiotherapy has side effects reduced by H 2 . ...
... By comparing pioglitazone-treated and control groups, the administration of water-rich hydrogen reduced the expression of PPARα, and hepatic cholesterol and improved antioxidative activity in the liver of a mouse model [7]. IL-6, TNF-α, inflammatory cytokines, apoptosis biomarkers, and oxidative stress were all strongly inhibited by hydrogen-rich water [43]. The effect of molecular hydrogen on cancer is summarized (Fig. 3). ...
Molecular hydrogen proved itself as a novel therapeutic candidate and has been thriving from the beginning with its potential clinical significance, higher affinity, and cellular integrity and permeability. Hydrogen Therapy (HT) has gained scientists' attention with the proven clinical ability to attenuate chronic inflammation, diminish oxidative stress, restrict apoptosis, minimize cellular injury, and refine tissue functioning. Therapeutic Implementation of H2 for disease prevention and treatment is a newly emerging field with limited knowledge available on formulations, tissue-specific effects, efficacy, and safety. This article will discuss HT's therapeutic potential for its efficacy and safety in cardiovascular, respiratory, hematological, metabolic, infectious, and neurodegenerative disorders. In addition to this, the molecular mechanisms and nanotechnological implications of hydrogen therapy will be discussed in detail. Finally, the article will provide insight into advancements and automation, future perspectives, and recommendations. There is a need to study and conduct higher-scale trials targeting personalized treatments under molecular and genetic vitals.
... By reducing hepatic oxidative stress, apoptosis, and inflammation, H 2 prevents progression of nonalcoholic steatohepatitis-related hepatocarcinogenesis [180]. However, a previous study has found that combining H 2 with platinum nanocolloids exerts carcinostatic and carcinocidal effects by increasing H 2 peroxide generation and cell death in a human gastric cancer cell line NUGC-4 [181]. ...
... human;[176] Sprague-Dawley rats, C57BL/6 mice, and db/db mice; and[177] Sprague-Dawley rats.Inhibits ROS, apoptosis, and inflammation in lesion tissue; downregulates chromosome 3; enhances anticancer effects; alleviates side effects of anticancer therapies; modulates immune function; and restores exhausted CD8+ T cells.[127] A549 and H1975 cells;[180] C57BL/6 mice;[182] mouse colon carcinoma cell line and BALB/c mice;[185] C57BL/6J mice and human lung cancer cell lines A549;[186] Wistar albino rats;[187] human; and[189] human. ...
Ageing is a physiological process of progressive decline in the organism function over time. It affects every organ in the body and is a significant risk for chronic diseases. Molecular hydrogen has therapeutic and preventive effects on various organs. It has antioxidative properties as it directly neutralizes hydroxyl radicals and reduces peroxynitrite level. It also activates Nrf2 and HO-1, which regulate many antioxidant enzymes and proteasomes. Through its antioxidative effect, hydrogen maintains genomic stability, mitigates cellular senescence, and takes part in histone modification, telomere maintenance, and proteostasis. In addition, hydrogen may prevent inflammation and regulate the nutrient-sensing mTOR system, autophagy, apoptosis, and mitochondria, which are all factors related to ageing. Hydrogen can also be used for prevention and treatment of various ageing-related diseases, such as neurodegenerative disorders, cardiovascular disease, pulmonary disease, diabetes, and cancer. This paper reviews the basic research and recent application of hydrogen in order to support hydrogen use in medicine for ageing prevention and ageing-related disease therapy.
... Excessive lipid droplet accumulation in NAFL patients can result in oxidative stress, inflammation, and hepatocyte injury, eventually leading to NASH [40]. Because NAFLD is a multifactorial disease, different mouse models are used to study NAFLD pathogenesis, including methionine-choline diet-induced NASH, high-fat diet-induced NAFLD, and streptozotocin-induced NASH-related hepatocarcinogenic models [27,[36][37][38]48]. Several of these studies used hydrogen therapy, which demonstrated a therapeutic effect against NAFLD (Table 1). ...
... Interestingly, HRW treatment also modulates transcriptional changes in lipid metabolic genes by increasing peroxisomal fatty acyl-CoA oxidase (ACOX), a rate-limiting enzyme for beta-oxidation of very-long-chain fatty acids. It also inhibits the transcription of CD36, a key protein in fatty acid uptake [38]. ...
Our body composition is enormously influenced by our lifestyle choices, which affect our health and longevity. Nutrition and physical activities both impact overall metabolic condition, thus, a positive energy balance causes oxidative stress and inflammation, hastening the development of metabolic syndrome. With this knowledge, boosting endogenous and exogenous antioxidants has emerged as a therapeutic strategy for combating metabolic disorders. One of the promising therapeutic inventions is the use of alkaline reduced water (ARW). Aside from its hydrating and non-caloric properties, ARW has demonstrated strong antioxidant and anti-inflammatory properties that can help stabilize physiologic turmoil caused by oxidative stress and inflammation. This review article is a synthesis of studies where we elaborate on the intra- and extracellular effects of drinking ARW, and relate these to the pathophysiology of common metabolic disorders, such as obesity, diabetes mellitus, non-alcoholic fatty liver disease, and some cancers. Highlighting the health-promoting benefits of ARW, we also emphasize the importance of maintaining a healthy lifestyle by incorporating exercise and practicing a balanced diet as forms of habit.
... На тваринних моделях було показано пригнічення росту пухлин під впливом H 2 через різні протипухлинні механізми. D. Kawai та ін., 2012 [30], також продемонстрували, що воднева вода має значну антиоксидантну та протиканцерогенну дію. Автори показали, що молекулярний водень знижує експресію ключових генів метаболізму ліпідів, впливаючи на печінкову експресію генів синтезу й окиснення жирних кислот через взаємодію з рецепторами, що активуються пероксисомними проліфераторами (PPAR). ...
РЕЗЮМЕ. Окислювальні процеси відіграють важливу роль в онкогенезі, в тому числі колоректального раку (КРР). Існує ряд повідомлень про зміни у функціонуванні системи циклічних нуклеотидів в онкологічних хворих, зокрема, щодо ролі цГМФ. Застосування антиоксидантів потенційно може уповільнити пухлинний процес. Ураховуючи антиоксидантні та протизапальні властивості молекулярного водню, актуальним є вивчення потенційної терапевтичної дії збагаченої воднем води при онкопатології. Мета – оцінити вплив води, насиченої молекулярним воднем, на вміст циклічного гуанозинмонофосфату та малонового діальдегіду в сироватці крові лікованих 5-фторурацилом (5ФУ) щурів із колоректальним раком. Матеріал і методи. Експерименти проведені на 90 щурах-самцях лінії Вістар. КРР моделювали за допомогою підшкірних ін’єкцій 1,2-диметилгідразину в дозі 7,2 мг/кг 1 раз на тиждень протягом 30 тижнів. Щури споживали воду, збагачену молекулярним воднем, у концентрації 0,6 ppm ad libitum. У сироватці крові тварин вміст цГМФ визначали методом імуноферментного аналізу, рівень МДА визначали колориметричним методом. Результати. Вміст цГМФ у сироватці крові щурів з КРР був на 39,9 % нижчим, ніж у тварин в контрольній групі, водночас рівень МДА був збільшений в 1,9 раза. Вміст цГМФ у сироватці крові тварин з КРР, що споживали збагачену молекулярним воднем воду, був на 38,1 % вищим, ніж у тварин, що споживали водопровідну воду, при цьому рівень МДА достовірно (на 30,8 %) знижувався. Введення 5ФУ тваринам з КРР призводило до зниження вмісту цГМФ у сироватці крові на 42,3 % і до збільшення МДА в 2,3 раза, порівняно з контролем, але ці показники достовірно не відрізнялися від аналогічних у тварин з КРР, які були не ліковані 5ФУ. Рівень цГМФ у сироватці крові щурів, які були ліковані 5ФУ та споживали воду, збагачену молекулярним воднем, був на 24,1 % вищим, ніж у лікованих 5ФУ щурів з КРР, що мали доступ до звичайної води. Воднева вода істотно не вплинула на інтенсивність окиснювального стресу у щурів з КРР, які отримували 5ФУ. Висновок. У щурів з КРР, індукованим 1,2-диметилгідразином, рівень цГМФ у сироватці крові знижується, а рівень МДА підвищується. Лікування щурів з КРР за допомогою 5ФУ не викликає достовірних змін інтенсивності окисного стресу та в системі циклічних нуклеотидів. Споживання води, насиченої молекулярним воднем, тваринами з КРР, приводить до значного покращання показників інтенсивності окиснювального стресу та системи циклічних нуклеотидів.
... Furthermore, cellular DNA is also highly susceptible to attack by ROS [66]. 8-OHDG content is a typical biomarker reflecting oxidative damage to DNA [67]. The intracellular levels of 8-OHDG were examined using an enzyme immunoassay, which revealed no significant change in the 8-OHDG content of the low NP-treated groups (10, 20, and 30 mg/L). ...
The improper disposal of plastic products/wastes can lead to the release of nanoplastics (NPs) into environmental media, especially soil. Nevertheless, their toxicity mechanisms in soil invertebrates remain unclear. This study investigated the impact of polystyrene NPs on Eisenia fetida (Savigny, 1826) immune cells, focusing on oxidative stress, immune responses, apoptosis, and necrosis. Results showed that 100 nm NPs were internalized into the cells, causing cytotoxicity. NPs were observed to inhibit cell viability by increasing reactive oxygen species, decreasing the levels of antioxidants (e.g., superoxide dismutase, catalase, and glutathione), and inducing lipid peroxidation and DNA oxidation. Additionally, assays on neutral red retention time, lysozyme activity, and Ca2⁺ levels demonstrated that NPs resulted in a loss of lysosomal membrane stability and a reduction in immune resistance. The depolarization of the mitochondrial membrane potential and the results of the apoptosis assays confirmed that the NPs induced the onset of early apoptosis. The difficulty of the NP in causing cell death by disrupting the plasma membrane was demonstrated by the results of the lactate dehydrogenase release assays in relation to cell necrosis. This research provides cellular-level insights into the ecological risks of NP exposure on soil fauna.
... For STZ-HFD induced HCC model, a mouse model mimicking nonalcoholic fatty liver disease-related HCC [19], control or early-Abx male mice was induced by a single subcutaneous injection of 200 μg Streptozocin (Sigma) at 2 days after birth and feeding with HFD (cat#: D12492, Rodent diet with 60 kcal% fat) after 4 weeks of age. At 5th month, the mice were euthanized by carbon dioxide inhalation followed by cervical dislocation. ...
Background
Metabolic dysfunction-associated steatotic liver disease (MASLD) asscociated hepatocellular carcinoma (HCC) is becoming a growing concern in global healthcare. The early-life gut microbiota plays a crucial role in maintaining healthy. However, the impact of early-life gut microbiota dysbiosis on the advancement of MASLD-HCC remains inadequately understood.
Methods
In the present study, we investigated the role of early-life gut microbiota in the development of MASLD-HCC in streptozotocin and high-fat diet (STZ-HFD) induced mouse model. We recorded the body weight and lifespan, and dynamically monitored the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (T-CHO) and blood glucose in the serum monthly. In addition, we examined various immune cells present in the liver, such as T cells, B cells, NK cells, NKT cells, αβT cells, γδT cells, macrophage and MDSC cells by flow cytometry and conducted liquid chromatography mass spectrometry (LC–MS) based analysis on liver tissue from control and early-life antibiotic exposure mice (early-Abx) MASLD-HCC mice.
Results
We found that early-Abx mice suffered from more severe tumor burden and further confirmed that hepatocytes and immune cells were all disturbed. Importantly, early-life antibiotic exposure alters the liver metabolic profiling especially glycerophospholipids and lipid accumulation. Furthermore, mice exposed to antibiotics in early-life showed disturbances in glucose metabolism and developed insulin resistance.
Conclusions
Collectively, our findings revealed that early-life antibiotic exposure accelerated the progression of MASLD-HCC by impairing the hepatocytes, immune homeostasis and metabolites persistently, highlighting the importance of the early-life microbiota in the development of MASLD-HCC.
... A recent meta-analysis examining NAFLD in Asia reported an incidence rate of 50.9 per 1000 person-years. Mainland China hepatocarcinogenesis [23]. Consumption of hydrogen-rich water not only improves highfat-diet-induced type 2 diabetes, but also NAFLD in mice due to its antioxidative and anti-inflammatory activities [24,25]. ...
The prevalence of non-alcoholic fatty liver disease (NAFLD) has dramatically increased in recent years, and it is highly associated with metabolic diseases, as well as the development of hepatocellular carcinoma. However, effective therapeutic strategies for the treatment of NAFLD are still scarce. Although hydrogen-rich water shows beneficial effects for hepatic steatosis, the inconvenience limits the application of this antioxidant. In light of this, hydrogen-rich coral calcium (HRCC) was developed due to its convenience and quantifiable characteristics. However, the effects of HRCC on NAFLD are still unknown. In the present study, we found that HRCC treatment improved methionine-and-choline-deficient diet (MCD)-induced hepatic steatosis, increased aspartate aminotransferase and alanine aminotransferase levels, and elevated hepatic inflammatory factor expressions in mice. In addition to the increased expressions of antioxidative enzymes, we found that HRCC increased the expressions of bile acid biosynthesis-related genes, including Cyp8b1 and Cyp27a1. Increased hepatic bile acid contents, such as muricholic acids, 23 nor-deoxycholic acid, glycoursodeoxycholic acid, and cholic acids, were also confirmed in MCD mice treated with HRCC. Since the biogenesis of bile acids is associated with the constitution of gut microbiome, the alterations in gut microbiome by HRCC were evaluated. We found that HRCC significantly changed the constitution of gut microbiome in MCD mice and increased the contents of Anaerobacterium, Acutalibacter, Anaerosacchariphilus, and Corynebacterium. Taken together, HRCC improved MCD-induced NAFLD through anti-inflammatory mechanisms and by increasing antioxidative activities. Additionally, HRCC might alter gut microbiome to change hepatic bile acid contents, exerting beneficial effects for the treatment of NAFLD.
... On the one hand, H 2 can inhibit the damage of cellular DNA by ·OH and ONOO-to prevent cancer development; on the other hand, H 2 can remove ROS from cancer cells and inhibit multiple ROS-dependent metabolic signaling pathways to suppress cancer development. Studies have confirmed that H 2 can effectively reduce oxidative stress caused by various pathological conditions, including cancers, and promote the restoration of redox homeostasis (Adzavon et al. 2022;Shi et al. 2016;Kawai et al. 2012). ...
Gas signaling molecules, including carbon monoxide (CO), nitric oxide (NO), and hydrogen sulfide (H2S), have been shown to have cancer therapeutic potential, pointing to a new direction for cancer treatment. In recent years, a series of studies have confirmed that hydrogen (H2), a weakly reductive gas, also has therapeutic effects on various cancers and can mitigate oxidative stress caused by radiation and chemotherapy, reducing tissue damage and immunosuppression to improve prognosis. Meanwhile, H2 also has immunomodulatory effects, inhibiting T cell exhaustion and enhancing T cell anti-tumor function. It is worth noting that human intestinal flora can produce large amounts of H2 daily, which becomes a natural barrier to maintaining the body’s resistance to diseases such as tumors. Although the potential anti-tumor mechanisms of H2 are still to be investigated, previous studies have shown that H2 can selectively scavenge highly toxic reactive oxygen species (ROS) and inhibit various ROS-dependent signaling pathways in cancer cells, thus inhibiting cancer cell proliferation and metastasis. The ROS scavenging ability of H2 may also be the underlying mechanism of its immunomodulatory function. In this paper, we review the significance of H2 produced by intestinal flora on the immune homeostasis of the body, the role of H2 in cancer therapy and the underlying mechanisms, and the specific application of H2 to provide new ideas for the comprehensive treatment of cancer patients.
... Sifat-sifat ini menjadikannya molekul yang menarik untuk dikaji dan perlu kami share ke masyarakat. Namun, memerlukan studi lebih lanjut untuk menjelaskan mekanisme molekuler dan dosis optimal untuk H2, sehingga penelitian pendahuluan pada hewan dan manusia perlu dilakukan (Kawai et al., 2012). Studi klinis minum AKH telah menunjukkan efek menguntungkan pada beberapa penyakit seperti penyakit Parkinson, diabetes tipe II, rheumatoid arthritis, miopati mitokondria, kelelahan otot, sindrom metabolik, hiperlipidemia, peradangan hati (hepatitis B), tumor hati dan lain-lain (Kang et al., 2011). ...
One form of activity in health promotion is in the form of socialization that we do. These activities strengthen the skills and abilities of individuals to take action, and the capacity of groups or communities to act collectively to exercise control over the determinants of health. Socialization activities strengthen the skills and abilities of individuals to take action, and the capacity of groups or communities to act collectively to exercise control over the determinants of health. This activity uses lecture, discussion and question and answer methods with residents. The location of the activity was Persil Village, Central Lombok with 25 participants and accompanied by 3 lecturers and 5 students. The results of our socialization show that local residents need solutions to health problems and hope that Hydrogen Rich Water (AKH) is a solution to the disease problems that they and their families are currently facing. The knowledge of the local community is still categorized as low and requires special attention from the local government and stakeholders. Some residents who participated in the activity had a history of their own diseases, such as gastritis, cholesterol, high blood pressure, allergies and some of their families also had a history of certain diseases, such as kidney stones, diabetes, stroke, cancer and so on. Persil villagers gained knowledge about health, especially the differences in various kinds of water consumed daily. The villagers originally thought that all water is the same, but after we provided information about water, the villagers became more understanding and more selective about the drinks they consume daily.
... de- creasing the number of tumor cells sharply. H 2 also downregulates proinflammatory cytokines ( e.g. , TNF-α and IL-6) to alleviate tumor inflammation [102] and also regulates micro-RNA ( e.g. , miR-21 and miR-199) to control tumor inflammation [103] . In terms of oxidative stress, H 2 inhibits vascular endothelial growth factor (VEGF) and ERK signaling downregulation by reducing excessive ROS [104] and activates the body's own antioxidant system through the production of endogenous antioxidants, such as catalase (CAT) and superoxide dismutase (SOD) [105] . ...
... 6 For instance, drinking hydrogen-rich water has been found to effectively reduce hepatic oxidative stress, apoptosis, inflammation, and hepatocarcinogenesis, making it a potential treatment for alcoholic hepatitis. 7 Supplementation of H 2 S in rectal cancer avoids the loss of resistance to 5-fluorouracil induced by cystine-glutamate antiporter interference, that suggesting a functional role for H 2 S in the maintenance of chemoresistance. 8 Gas molecules play an important role in various pathophysiological states, which are of great significance for clinical treatment. ...
Medical gases play an important role in the pathophysiology of human diseases and have received extensive attention for their role in neuroprotection. Common pathological mechanisms of spinal cord injury include excitotoxicity, inflammation, cell death, glial scarring, blood-spinal cord barrier disruption, and ischemia/reperfusion injury. Nitric oxide and hydrogen sulfide are important gaseous signaling molecules in living organisms; their pathological role in spinal cord injury models has received more attention in recent years. This study reviews the possible mechanisms of spinal cord injury and the role of nitric oxide and hydrogen sulfide in spinal cord injury.
... Therefore, the effects of antioxidants on cancer are two-sided, either inhibiting or promoting carcinogenesis, depending on the conditions. H2 has not yet been shown to exert carcinogenic effects, whereas its carcinogenic inhibitory effects have been demonstrated [65]. ...
With the rapid increase in diabetes worldwide, the number of patients with diabetic kidney disease (DKD), a complication of diabetes mellitus, is also on the rise. DKD is a major cause of chronic kidney disease progressing to end-stage renal failure; however, current medical treatments for DKD do not achieve satisfactory outcomes. Molecular hydrogen (H2) is an antioxidant that selectively reduces hydroxyl radicals, a reactive oxygen species with a very potent oxidative capacity. H2 was recently shown to exert not only antioxidant, but also anti-inflammatory, cell lethality-regulating, and signal transduction-regulating effects, and is now being applied clinically. Many factors contribute to the onset and progression of DKD, with mitochondrial dysfunction, oxidative stress, and inflammation being strongly implicated. Recent preclinical and clinical trials reported that substances with antioxidant properties may slow the progression of DKD. Therefore, we conducted a literature review on animal models and human clinical trials in which H2 showed efficacy against various renal diseases. This literature review and our previous findings collectively suggest that H2 exerts therapeutic effects in patients with DKD by improving mitochondrial function. Future large-scale clinical studies are needed to confirm these findings.
... Treatment with H2 could be prospectively used in agriculture to reduce postharvest senescence [22]. In a mouse model of NASH, hepatic mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-6, fatty acid synthesis-related genes, and peroxisome proliferator-activated receptor-α decreased significantly after treatment with hydrogen-rich water (HRW), and the oxidative stress biomarker 8-hydroxydeoxyguanosine in the liver tissue was reduced [23]. In a high-fat diet-induced NAFLD rat model, treatment with H2 reduced body weight gain, improved glucose and lipid metabolism, attenuated hepatic steatosis, and improved hepatic mitochondrial dysfunction [24]. ...
Molecular hydrogen (H2) has been recognized as a novel medical gas with antioxidant and anti-inflammatory effects. Non-alcoholic fatty liver disease (NAFLD) is a liver pathology with increased fat accumulation in liver tissue caused by factors other than alcohol consumption. Platelet mitochondrial function is considered to reflect systemic mitochondrial health. We studied the effect of adjuvant therapy with hydrogen-rich water (HRW) on coenzyme Q10 (CoQ10) content and platelet mitochondrial bioenergetics in patients with NAFLD. A total of 30 patients with NAFLD and 15 healthy volunteers were included in this clinical trial. A total of 17 patients (H2 group) drank water three × 330 mL/day with tablets producing HRW (>4 mg/L H2) for 8 weeks, and 13 patients (P group) drank water with placebo tablets producing CO2. The concentration of CoQ10-TOTAL was determined by the HPLC method, the parameter of oxidative stress, thiobarbituric acid reactive substances (TBARS), by the spectrophotometric method, and mitochondrial bioenergetics in platelets isolated from whole blood by high-resolution respirometry. The patients with NAFLD had lower concentrations of CoQ10-TOTAL in the blood, plasma, and platelets vs. the control group. Mitochondrial CI-linked LEAK respiration was higher, and CI-linked oxidative phosphorylation (OXPHOS) and CII-linked electron transfer (ET) capacities were lower vs. the control group. Plasma TBARS concentrations were higher in the H2 group. After 8 weeks of adjuvant therapy with HRW, the concentration of CoQ10 in platelets increased, plasma TBARS decreased, and the efficiency of OXPHOS improved, while in the P group, the changes were non-significant. Long-term supplementation with HRW could be a promising strategy for the acceleration of health recovery in patients with NAFLD. The application of H2 appears to be a new treatment strategy for targeted therapy of mitochondrial disorders. Additional and longer-term studies are needed to confirm and elucidate the exact mechanisms of the mitochondria-targeted effects of H2 therapy in patients with NAFLD.
... The various routes of administration, application and mechanisms of action of H 2 molecules in cancer treatment are listed in Table 1. Reversal of renal toxicity due to cisplatin Inhibition of apoptosis and ROS [91] suppression of tumor growth Arrest and induction of apoptosis [92] H 2 -rich water Improvement of mFOLFOX6 regimen-induced liver toxicity Inhibition of oxidative stress [93] 44Gy electronic beam reversal of skin damage created Inflammatory cytokines and oxidative stress reduction [94] inhibition of cancer stem cells Inhibition of angiogenesis [92] Prevention of gefitinib-induced lung injury Cytokines inflammatory and oxidative stress reduction [95] Prevention of cisplatin-induced nephrotoxicity Elimination of oxygen radicals [73] Reversal of mortality and body-weight loss caused by cisplatin Inhibition of ROS [73] Incidence of tumors and suppression of growth Inhibition of inflammatory cytokines and oxidative stress, Induction of apoptosis [96] Improved quality of life Action of antioxidants [73] ...
Age-related diseases represent the largest threat to public health. Aging is a degenerative, systemic, multifactorial and progressive process, coupled with progressive loss of function and eventually leading to high mortality rates. Excessive levels of both pro- and anti-oxidant species qualify as oxidative stress (OS) and result in damage to molecules and cells. OS plays a crucial role in the development of age-related diseases. In fact, damage due to oxidation depends strongly on the inherited or acquired defects of the redox-mediated enzymes. Molecular hydrogen (H2) has recently been reported to function as an anti-oxidant and anti-inflammatory agent for the treatment of several oxidative stress and aging-related diseases, including Alzheimer’s, Parkinson’s, cancer and osteoporosis. Additionally, H2 promotes healthy aging, increases the number of good germs in the intestine that produce more intestinal hydrogen and reduces oxidative stress through its anti-oxidant and anti-inflammatory activities. This review focuses on the therapeutic role of H2 in the treatment of neurological diseases. This review manuscript would be useful in knowing the role of H2 in the redox mechanisms for promoting healthful longevity.
... A common effect also seen in animal models is the suppression of tumour growth through various antitumor mechanisms. Kawai et al., (2012) noticed that hydrogen water could induce great antioxidative and anti-hepatic carcinogenic effects. In their study, H2 reduces the expression of key genes involved in lipid metabolism to control the hepatic expression of fatty acid synthesis and fatty acid oxidation genes by interacting with the peroxisome proliferator-activated receptors (PPAR) pathway ( Figure 2). ...
Background:
Cancer remains a challenging target to cure, with present therapeutic methods unable to exhibit restorative outcomes without causing severe negative effects. Molecular hydrogen (H2) has been reported to be a promising adjunctive therapy for cancer treatment, having the capability to induce anti-proliferative, anti-oxidative, pro-apoptotic and anti-tumoural effects. This review summarises the findings from various articles on the mechanism, treatment outcomes, and overall effectiveness of H2 therapy on cancer management.
Methods:
Using Cochrane, PubMed, and Google Scholar as the search engines, full-text articles in the scope of the study, written in English and within 10 years of publication were selected.
Results:
Out of the 677 articles, 27 articles fulfilled the eligibility criteria, where data was compiled into a table, outlining the general characteristics and findings. Throughout the different forms of H2 administration, study design and types of cancers reported, outcomes were found to be consistent.
Conclusion:
From our analysis, H2 plays a promising therapeutic role as an independent therapy as well as an adjuvant in combination therapy, resulting in an overall improvement in survivability, quality of life, blood parameters, and tumour reduction. Although more comprehensive research is needed, given the promising outcomes, H2 is worth considering for use as a complement to current cancer therapy.
... 20 In terms of metabolic diseases, that is, NAFLD, molecular hydrogen was proved to be effective in preventing liver fat accumulation in a HepG2 cell model 21 and animal models. [22][23][24] A randomized controlled pilot study was reported by Korovljev et al. that hydrogen-rich water improved mild-to-moderate NAFLD. 25 Though the number of cases in this study was limited, a prospect of molecular hydrogen as an adjuvant treatment is promising. ...
Non‐alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide with increasing incidence consistent with obesity, type 2 diabetes and cardiovascular diseases. No approved medication was currently available for NAFLD treatment. Molecular hydrogen (H2), an anti‐oxidative, anti‐inflammatory biomedical agent is proved to exhibit therapeutic and preventive effect in various diseases. The purpose of this study was to investigate the effect of hydrogen/oxygen inhalation on NAFLD subjects and explore the mechanism from the perspective of hepatocyte autophagy. We conducted a randomized, placebo‐controlled clinical trial of 13‐week hydrogen/oxygen inhalation (China Clinical Trial Registry [#ChiCTR‐IIR‐16009114]) including 43 subjects. We found that inhalation of hydrogen/oxygen improved serum lipid and liver enzymes. Significantly improved liver fat content detected by ultrasound and CT scans after hydrogen/oxygen inhalation was observed in moderate–severe cases. We also performed an animal experiment based on methionine and choline‐deficient (MCD) diet‐induced mice model to investigate effect of hydrogen on mouse NASH. Hydrogen/oxygen inhalation improved systemic inflammation and liver histology. Promoted autophagy was observed in mice inhaled hydrogen/oxygen and treatment with chloroquine blocked the beneficial effect of hydrogen. Moreover, molecular hydrogen inhibited lipid accumulation in AML‐12 cells. Autophagy induced by palmitic acid (PA) incubation was further promoted by 20% hydrogen incubation. Addition of 3‐methyladenine (3‐MA) partially blocked the inhibitory effect of hydrogen on intracellular lipid accumulation. Collectively, hydrogen/oxygen inhalation alleviated NAFLD in moderate–severe patients. This protective effect of hydrogen was possibly by activating hepatic autophagy.
... Kawai ve ark. 2012 [22], hidrojenle zenginleştirilmiş su (HRW) ve pioglitazon (PGZ) ilacının farelerde yağlı karaciğer iltihaplanma hastalığı olan Alkolsüz steatohepatit (NASH) ilerlemesi üzerindeki etkilerini incelemişlerdir. Oksidatif stres biyobelirteçleri, interlökin-6 ve tümör nekroz faktörü-α gibi inflamatuar faktörler ve apoptoz HRW ve PGZ gruplarında azalma göstermiştir. ...
ABSTRACT:
Molecular hydrogen, which has the H2 formula, is a colorless, odorless, tasteless, non-metallic and non-toxic gas. Molecular hydrogen has a high diffusion rate and spreads rapidly in biological tissues and cells. Molecular hydrogen does not change the body's redox reactions and does not cause any side effects. H2 is a selective antioxidant due to its ability to neutralize strong oxidants such as hydroxyl radicals in cells. This case increases the potential use of molecular hydrogen for preventative and therapeutic applications. In addition, by regulating various gene expressions, H2 exhibits anti-inflammatory and anti-apoptotic properties. At the same time, unlike the drugs used in the treatment of diseases, H2 penetrates the cell membrane easily. Inhaling hydrogen gas, drinking hydrogen water, injection of hydrogen-enriched saline, bathing with a hydrogen-rich water, and hydrogen-rich eye drops are the common methods of molecular hydrogen applications in the health field. Numerous studies on the biological and medical benefits of hydrogen have been carried out up to date, and research is still ongoing. As a result; due to its beneficial effects on the body and the absence of side effects, molecular hydrogen can have promising potential applications against many diseases.
ÖZET:
H2 formülüne sahip olan moleküler hidrojen, renksiz, kokusuz, tatsız, metalik olmayan ve toksik olmayan bir gazdır. Moleküler hidrojen (H2), yüksek difüzyon hızına sahiptir ve biyolojik doku ve hücrelerde hızla yayılır. Moleküler hidrojen, vücudun redoks reaksiyonlarını değiştirmemekte ve hiçbir yan etki göstermemektedir. Hücrelerdeki hidroksil radikal gibi güçlü oksidanları inhibe etme özelliğinden dolayı seçici bir antioksidandır. Bu durum, moleküler hidrojenin önleyici ve tedavi edici uygulamalar için potansiyelini arttırmaktadır. Ek olarak, çeşitli gen ifadelerini düzenleyerek, antiinflamatuar ve antiapoptik özellik sergilemektedir. Aynı zamanda hastalıkların tedavisinde kullanılan ilaçların aksine moleküler hidrojen, hücre zarına kolaylıkla nüfuz etmektedir. Hidrojen gazı solumak, hidrojenle zenginleştirilmiş su içmek, hidrojenle zenginleştirilmiş salin enjeksiyonu, hidrojenli su banyosu yapmak ve hidrojenli göz damlaları kullanmak sağlık alanında moleküler hidrojen uygulamasının yaygın yöntemleridir. Bugüne kadar hidrojenin biyolojik ve tıbbi faydaları üzerine çok sayıda çalışma yapılmıştır ve araştırmalar halen devam etmektedir. Sonuç olarak; vücuttaki olumlu etkileri, yan etkisinin bulunmaması sebebiyle, moleküler hidrojen birçok hastalığa karşı umut verici bir potansiyele sahiptir.
... Hw can be produced by injecting hydrogen gas [23,24], by generating hydrogen through electrolysis [14], and by generating hydrogen through the reaction of metal magnesium and water [25,26]. the solubility of hydrogen is approximately 1.6 ppm [27], and hydrogen concentrations of 0.8 ppm [28] and 0.3-0.4 ...
Hydrogen-rich water (HW) has been suggested to possess antioxidant properties of value in treatments of lifestyle diseases and for prevention of latent pathologies. To date, the potential benefits of HW against the deleterious effects of excessive salt intake and hypertension have not been investigated. Here, we first examined the effects of HW or HW supplemented with 0.1% ascorbic acid (HWA) on spontaneously hypertensive rats (SHR) that had been fed a normal diet. In comparison to control rats given distilled water (DW), we found that HW did not significantly influence systolic blood pressure (SBP) or diastolic blood pressure (DBP) in SHR; however, the increase in SBP and DBP were inhibited in the HWA group. Next, four groups of SHR were given DW, 0.1% ascorbic acid-added DW (DWA), HW, or HWA in combination with a 4% NaCl-added diet. SHR fed the 4% NaCl-added diet showed increased hypertension; HWA treatment resulted in a significant reduction in blood pressure. The HWA group tended to have lower plasma angiotensin II levels than the DW group. In addition, urinary volumes and urinary sodium levels were significantly lower in the HWA group than the DW group. Urinary isoprostane, an oxidative stress marker, was also significantly lower in the HWA group, suggesting that the inhibitory effect of HWA on blood pressure elevation was caused by a reduction in oxidative stress. These findings suggest a synergistic interaction between HW and ascorbic acid, and also suggest that HWA ingestion has potential for prevention of hypertension.
... Kawai ve ark. 2012 [22], hidrojenle zenginleştirilmiş su (HRW) ve pioglitazon (PGZ) ilacının farelerde yağlı karaciğer iltihaplanma hastalığı olan Alkolsüz steatohepatit (NASH) ilerlemesi üzerindeki etkilerini incelemişlerdir. Oksidatif stres biyobelirteçleri, interlökin-6 ve tümör nekroz faktörü-α gibi inflamatuar faktörler ve apoptoz HRW ve PGZ gruplarında azalma göstermiştir. ...
H2 formülüne sahip olan moleküler hidrojen, renksiz, kokusuz, tatsız, metalik olmayan ve toksik olmayan bir gazdır. Moleküler hidrojen (H2), yüksek difüzyon hızına sahiptir ve biyolojik doku ve hücrelerde hızla yayılır. Moleküler hidrojen, vücudun redoks reaksiyonlarını değiştirmemekte ve hiçbir yan etki göstermemektedir. Hücrelerdeki hidroksil radikal gibi güçlü oksidanları inhibe etme özelliğinden dolayı seçici bir antioksidandır. Bu durum, moleküler hidrojenin önleyici ve tedavi edici uygulamalar için potansiyelini arttırmaktadır. Ek olarak, çeşitli gen ifadelerini düzenleyerek, antiinflamatuar ve antiapoptik özellik sergilemektedir. Aynı zamanda hastalıkların tedavisinde kullanılan ilaçların aksine moleküler hidrojen, hücre zarına kolaylıkla nüfuz etmektedir. Hidrojen gazı solumak, hidrojenle zenginleştirilmiş su içmek, hidrojenle zenginleştirilmiş salin enjeksiyonu, hidrojenli su banyosu yapmak ve hidrojenli göz damlaları kullanmak sağlık alanında moleküler hidrojen uygulamasının yaygın yöntemleridir. Bugüne kadar hidrojenin biyolojik ve tıbbi faydaları üzerine çok sayıda çalışma yapılmıştır ve araştırmalar halen devam etmektedir. Sonuç olarak; vücuttaki olumlu etkileri, yan etkisinin bulunmaması sebebiyle, moleküler hidrojen birçok hastalığa karşı umut verici bir potansiyele sahiptir.
... At the stage of 2009-2012, many studies about drinking H 2 -rich water (hydrogen water) and injecting H 2 -rich saline began to appear, 9,10 and there were also studies to increase the supplementation of H 2 through intestinal bacteria. 11 That was a period when scientists were discovering and using different H 2 -intake methods to testify the medical effects. ...
Gas medicine, including O 2 , NO, H 2 S, CO, CH 4 , has played important roles in prevention and treatment of diseases for a long time. Molecular hydrogen (H 2 ), the smallest diatomic molecule in nature, has become a rising star in gas medicine in the past decades. Many studies have shown that H 2 has preventive and therapeutic effects on various diseases through its selective antioxidant activity. H 2 , as a non-toxic gas for the human body and convenience to obtain, has provided a great possibility to be used widely. Currently, the main difficulties in hydrogen medicine are lack of definitive clinical evidence and the molecular basis of hydrogen effects. In this paper, the authors have conducted a comprehensive review and analysis of these issues, and also proposed the possibility of developing Hydrogen Biology and Hydrogen Medicine as new disciplines of biology and medicine.
... Many models of severe inflammation such as the MCD diet model, cholesterol and cholate model, and PTEN null mice do not develop insulin resistance. Nonetheless, they are considered useful for evaluating NASH histopathology and the effects of various substances on NASH [50][51][52]. Therefore, the present model is thought to be a robust model for analyzing the histopathology of NASH, but it may not be appropriate for analyzing the pathophysiology of the disease. ...
... High-content hydrogen water (HHW) is a powerful antioxidant that can alleviate the progression of non-alcoholic steatohepatitis (NASH) [190]. In a study by Wang et al., the effect of HHW on the HFD model was investigated [172]. ...
Background
It has been suggested that dysregulation of long non-coding RNAs (lncRNAs) could be associated with the incidence and development of metabolic disorders.Aim Accordingly, this narrative review described the molecular mechanisms of lncRNAs in the development of metabolic diseases including insulin resistance, diabetes, obesity, non-alcoholic fatty liver disease (NAFLD), cirrhosis, and coronary artery diseases (CAD). Furthermore, we investigated the up-to-date findings on the association of deregulated lncRNAs in the metabolic disorders, and potential use of lncRNAs as biomarkers and therapeutic targets.Conclusion
LncRNAs/miRNA/regulatory proteins axis plays a crucial role in progression of metabolic disorders and may be used in development of
therapeutic and diagnostic approaches.
... 27 The genes of oxidoreduction-related proteins, including hydroxymethylglutaryl co-enzyme-A reductase, were significantly enriched in the livers of rats after drinking HRW. 28 Hepatocyte death induced by antimycin A was significantly suppressed after culturing in hydrogen-rich medium. However, this did not suppress hepatic stellate cell activation. ...
Hydrogen therapy is a very promising treatment against several diseases due to its mild attributes, high affinity and inherent biosafety. However, there is little elaboration about current hydrogen treatment in liver diseases. This article introduces the administration of hydrogen and mechanisms of hydrogen therapy in vivo, including modulating reactive oxygen species, apoptosis and autophagy, and inflammation, affecting mitochondria, as well as protein transporters. The major focus is clinical hydrogen use and related mechanisms in liver dysfunction or diseases, including non-alcoholic fatty liver disease, hepatitis B, liver dysfunction caused by liver tumour and colorectal tumour chemotherapy. Further, the article reveals ex vivo hydrogen application in liver protection. Finally, the article discusses the current and future challenges of hydrogen therapy in liver diseases, aiming to improve knowledge of hydrogen therapy and provide some insights into this burgeoning field.
... It can be absorbed into the blood circulation, such that it reaches the target organ either by blood circulation or free diffusion (11). Therefore, treatments involving exogenous H 2 , including breathing H 2 gas, injection with H 2 -rich saline and drinking H 2 -rich water, may protect against excessive oxidative stress-and inflammation-related liver damage, including liver injury induced by drugs, sepsis, bile duct ligation, ischemia/reperfusion (I/R), CO 2 pneumoperitoneum and chronic intermittent hypoxia, in addition to non-alcoholic fatty liver disease (NAFLD) (12)(13)(14)(15)(16)(17). Furthermore, drinking H 2 -rich water has been indicated to protect against chronic ethanol-induced hepatotoxicity (18). ...
Binge alcohol drinking is fast becoming a global health concern, with the liver among the first organ involved and the one afflicted with the greatest degree of injury. Oxidative stress, alterations in hepatic metabolism, immunity and inflammation have all been reported to contribute to the development of alcoholic liver disease (ALD). Hydrogen gas (H2) serves a key role in the modulation of hepatic redox, immune and inflammatory homeostasis. However, the effects of treatment using intraperitoneal injection of H2 on ALD remain unexplored. Therefore, the aim of the present study was to investigate the effects and underlying mechanism of intraperitoneal injection of H2 on acute alcohol-induced liver injury in a mouse model. H2 was administered by daily intraperitoneal injections (1.0 ml/100 g) for 4 days. On day 4, the mice received H2 after fasting for 5.5 h. After 30 min, the mice were administered with 33% (v/v) ethanol at a cumulative dose of 4.5 g/kg body weight by four equally divided gavages at 20-min intervals. Blood and liver tissues were collected at 16 h after the first ethanol gavage. Subsequently, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride and total cholesterol (TC) levels were analyzed using an Automatic Clinical Analyzer. Hepatic JNK activity and GAPDH levels were examined by western blotting. It was observed that acute ethanol gavage induced liver injury, as indicated by significantly increased serum ALT and AST levels, which were effectively decreased by H2 at 16 h after the first ethanol gavage. In addition, H2 treatment reduced serum TC levels in the Alcohol+H2 group when compared with those in Alcohol group. Mechanistically, H2 attenuated hepatic JNK phosphorylation induced by acute ethanol gavage. Therefore, the results of the present study demonstrated that treatment with exogenous H2 by intraperitoneal injection may alleviate acute alcohol-induced liver injury by inhibiting hepatic JNK activation, which may represent a novel therapeutic strategy for ALD.
... In mice, H 2 -rich water appears to prevent the evolution of hepatocarcinogenesis induced in mice. It was suggested that these positive outcomes were partly due to a reduction in CSCs [55] Recent experimental results indicate that H 2 is an antitumor agent in the treatment of glioblastoma. The invasive properties of glioblastoma are a key problem for curative treatment if a surgical resection is impracticable. ...
Simple Summary
Protons (H⁺) and molecular hydrogen (H2) in the cell are critical in a wide variety of processes. New cancer treatment uses H2, a biologically inactive gas. H2 can rapidly penetrate cell membranes and reach subcellular components to protect nuclear DNA and mitochondria. H2 reduces oxidative stress, exerts anti-inflammatory effects, and acts as a modulator of apoptosis. Exogenous H2 is a protective therapy that can be used in cancer. Cyclotrons and synchrotrons are currently used to produce protons. Proton beam radiotherapy (PBT) offers great promise for the treatment of a wide variety of cancers. H2 and different types of H2 donors may represent a novel therapeutic strategy in cancer treatment.
Abstract
Understanding the structure and dynamics of the various hydrogen forms has been a subject of numerous studies. Protons (H⁺) and molecular hydrogen (H2) in the cell are critical in a wide variety of processes. A new cancer treatment uses H2, a biologically inactive gas. Due to its small molecular weight, H2 can rapidly penetrate cell membranes and reach subcellular components to protect nuclear DNA and mitochondria. H2 reduces oxidative stress, exerts anti-inflammatory effects, and acts as a modulator of apoptosis. Exogenous H2, administered by inhalation, drinking H2-rich water, or injecting H2-rich saline solution, is a protective therapy that can be used in multiple diseases, including cancer. In particle therapy, cyclotrons and synchrotrons are the accelerators currently used to produce protons. Proton beam radiotherapy (PBT) offers great promise for the treatment of a wide variety of cancers due to the sharp decrease in the dose of radiation at a defined point. In these conditions, H2 and different types of H2 donors may represent a novel therapeutic strategy in cancer treatment.
... Many models of severe inflammation such as the MCD diet model, cholesterol and cholate model, and PTEN null mice do not develop insulin resistance. Nonetheless, they are considered useful for evaluating NASH histopathology and the effects of various substances on NASH [50][51][52]. Therefore, the present model is thought to be a robust model for analyzing the histopathology of NASH, but it may not be appropriate for analyzing the pathophysiology of the disease. ...
Several recent experimental studies have investigated the effects of caffeine and chlorogenic acid (CGA), representative ingredients of coffee, on nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH). However, the results are conflicting, and their effects are yet to be clarified. In the present study, we examined the effects of caffeine and CGA on choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD)-fed mice, relatively new model mice of NASH. Seven-week-old male C57BL/6J mice were divided into the following groups: Control diet (control), CDAHFD (CDAHFD), CDAHFD supplemented with 0.05% (w/w) caffeine (caffeine), and CDAHFD supplemented with 0.1% (w/w) CGA (CGA). After seven weeks, the mice were killed and serum biochemical, histopathological, and molecular analyses were performed. Serum alanine aminotransferase (ALT) levels were significantly higher in the caffeine and CGA groups than in the CDAHFD group. On image analysis, the prevalence of Oil red O-positive areas (reflecting steatosis) was significantly higher in the caffeine group than in the CDAHFD group, and that of CD45R-positive areas (reflecting lymphocytic infiltration) in the hepatic lobule was significantly higher in the caffeine and CGA groups than in the CDAHFD group. Hepatic expression of interleukin (IL)-6 mRNA was higher in the caffeine and CGA groups than in the CDAHFD group, and the difference was statistically significant for the caffeine group. In conclusion, in the present study, caffeine and CGA significantly worsened the markers of liver cell injury, inflammation, and/or steatosis in NASH lesions in mice.
... In a previous study, drinking H 2 -rich water was shown to slightly decrease the levels of hepatic cholesterol and triglycerides without any statistical significance in a methionine-choline-deficient diet-induced nonalcoholic steatohepatitis mouse model (53). The present study revealed the dose-dependent effect of H 2 inhalation on the alleviation of liver lipid accumulation and 66% H 2 inhalation was found to significantly decrease hepatic cholesterol and triglycerides. ...
Hydrogen exhibits therapeutic and preventive effects against various diseases. The present study investigated the potential protective effect and dose‑dependent manner of hydrogen inhalation on high fat and fructose diet (HFFD)‑induced nonalcoholic fatty liver disease (NAFLD) in Sprague‑Dawley rats. Rats were randomly divided into four groups: i) Control group, regular diet/air inhalation; ii) model group, HFFD/air inhalation; iii) low hydrogen group, HFFD/4% hydrogen inhalation; and iv) high hydrogen group, HFFD/67% hydrogen inhalation. After a 10‑week experiment, hydrogen inhalation ameliorated weight gain, abdominal fat index, liver index and body mass index of rats fed with HFFD and lowered the total area under the curve in an oral glucose tolerance test. Hydrogen inhalation also ameliorated the increase in liver lipid content and alanine transaminase and aspartate transaminase activities. Liver histopathologic changes evaluated with hematoxylin and eosin as well as Oil Red O staining revealed lower lipid deposition in hydrogen inhalation groups, consistent with the decrease in the expression of the lipid synthesis gene SREBP‑1c. The majority of the indicators were affected following treatment with hydrogen in a dose‑dependent manner. In conclusion, hydrogen inhalation may play a protective role by influencing the general state, lipid metabolism parameters, liver histology and liver function indicators in the rat model of metabolic syndrome with NAFLD.
We describe small heterojunction polymer dots (Pdots) with deep‐red light catalyzed H2 generation for diabetic skin wound healing. The Pdots with donor/acceptor heterojunctions showed remarkably enhanced photocatalytic activity as compared to the donor or acceptor nanoparticles alone. We encapsulate the Pdots and ascorbic acid into liposomes to form Lipo‐Pdots nanoreactors, which selectively scavenge •OH radicals in live cells and tissues under 650 nm light illumination. The antioxidant capacity of the heterojunction Pdots is ~10 times higher than that of the single‐component Pdots described previously. Under a total light dose of 360 J/cm2, the Lipo‐Pdots nanoreactors effectively scavenged •OH radicals and suppressed the expression of pro‐inflammatory cytokines in skin tissues, thereby accelerating the healing of skin wounds in diabetic mice. This study provides a feasible solution for safe and effective treatment of diabetic foot ulcers.
We describe small heterojunction polymer dots (Pdots) with deep‐red light catalyzed H2 generation for diabetic skin wound healing. The Pdots with donor/acceptor heterojunctions showed remarkably enhanced photocatalytic activity as compared to the donor or acceptor nanoparticles alone. We encapsulate the Pdots and ascorbic acid into liposomes to form Lipo‐Pdots nanoreactors, which selectively scavenge ⋅OH radicals in live cells and tissues under 650 nm light illumination. The antioxidant capacity of the heterojunction Pdots is ~10 times higher than that of the single‐component Pdots described previously. Under a total light dose of 360 J/cm², the Lipo‐Pdots nanoreactors effectively scavenged ⋅OH radicals and suppressed the expression of pro‐inflammatory cytokines in skin tissues, thereby accelerating the healing of skin wounds in diabetic mice. This study provides a feasible solution for safe and effective treatment of diabetic foot ulcers.
Molecular hydrogen (H2, dihydrogen) is an innovative experimental agent that could foster beneficial effects in a plethora of human conditions, including liver disorders. Recent trials demonstrated the positive effects of dihydrogen in several acute and chronic liver diseases, including chronic hepatitis B, non-alcoholic fatty liver disease, and liver cancer. Still, no systematic review or meta-analysis investigated the effects of dihydrogen intake on the hepatic function panel. For this report, we searched three relevant databases (PubMed, Web of Science, Scopus) from inception until December 24, 2022, using PRISMA guidelines. A literature search yielded 365 publications in total. After removing duplicates and studies that did not meet the inclusion criteria, 12 studies (published from 2008 to 2022) were included in this analysis. We found that serum malondialdehyde levels were significantly decreased following dihydrogen intake (pooled standardized mean difference = − 0.97 [95% confidence interval [CI], from − 1.65 to − 0.19; P = 0.01), with results indicating a large effect of dihydrogen intervention. A strong trend for a reduction in the liver function tests (including aspartate aminotransferase and alanine transaminase) has also been observed after dihydrogen administration (P < 0.20). Our findings suggested that dihydrogen can favorably affect the hepatic function panel; additional well-sampled interventional trials are highly warranted to corroborate our results.
As diabetes rates surge globally, there is a corresponding rise in the number of patients suffering from diabetic kidney disease (DKD), a common complication of diabetes. DKD is a significant contributor to chronic kidney disease, often leading to end-stage renal failure. However, the effectiveness of current medical treatments for DKD leaves much to be desired. Molecular hydrogen (H2) is an antioxidant that selectively reduces hydroxyl radicals, a reactive oxygen species with a very potent oxidative capacity. Recent studies have demonstrated that H2 not only possesses antioxidant properties but also exhibits anti-inflammatory effects, regulates cell lethality, and modulates signal transduction. Consequently, it is now being utilized in clinical applications. Many factors contribute to the onset and progression of DKD, with mitochondrial dysfunction, oxidative stress, and inflammation being strongly implicated. Recent preclinical and clinical trials reported that substances with antioxidant properties may slow the progression of DKD. Hence, we undertook a comprehensive review of the literature focusing on animal models and human clinical trials where H2 demonstrated effectiveness against a variety of renal diseases. The collective evidence from this literature review, along with our previous findings, suggests that H2 may have therapeutic benefits for patients with DKD by enhancing mitochondrial function. To substantiate these findings, future large-scale clinical studies are needed.
Вступ. У статті представлено огляд наукової літератури щодо впливу молекулярного водню, залежно від шляхів введення, на різні рівні організації організму при різних патологічних станах. Коротко подано історію відкриття молекулярного водню як біологічного агента і становлення водневої біомедицини. Охарактеризовано молекулярні, клітинні та системні аспекти біологічної дії молекулярного водню. Розкрито вплив молекулярного водню на різні пули клітин і процеси регенерації, його антиоксидантні та антиапоптичні ефекти. Описано протизапальну дію і регуляцію піроптозу молекулярним воднем. У цій статті інформацію проаналізовано з використанням баз даних PubMed, PubChem, ScienceDirect, Європейської Фармакопеї та наукової літератури. Мета дослідження – проаналізувати наукову літературу і систематизувати інформацію щодо уявлень про природу та шляхи надходження молекулярного водню в організм, особливостей і механізмів його молекулярних, клітинних, органно-тканинних та системних біологічних ефектів. Висновки. Молекулярний водень – найлегший і найпоширеніший медичний газ, який має широкий спектр біологічної активності й характеризується антиоксидантною, протизапальною та антиапоптичною діями. Він також бере участь у регуляції експресії численних генів, захисті біомакромолекул від окиснювального ушкодження, стимуляції виробництва енергії (АТФ) тощо. Водночас, незважаючи на різке збільшення кількості досліджень і публікацій щодо біомедичного застосування молекулярного водню, питання його використання як прорегенеративного агента потребує додаткового вивчення. Застосування даної молекули має численні переваги завдяки широкому спектру молекулярних реакцій, які вона викликає. Тому проведення цілеспрямованих досліджень у цій галузі може відкрити нові горизонти регенеративної медицини та створити інноваційну технологію прискореного відновлення організму.
Magnesium is generally known to degrade in aqueous environments by an electrochemical reaction. The corrosion products of magnesium include hydrogen gas, Mg2+, and Mg(OH)2. Here, we summarize the published literature describing the corrosion characteristics of magnesium, and the antitumor properties of magnesium-associated corrosion products, aiming to induce the therapeutic properties of magnesium and magnesium alloys in solid tumors. The therapeutic potential of corrosion products of magnesium is enormous. Hydrogen gas exhibits antioxidant and anti-inflammatory properties, which amount to potential anti-tumor characteristics. Mg(OH)2, which creates a localized alkaline microenvironment, represents a second approach for anti-tumor therapy with magnesium metal. Upregulated concentrations of Mg2+ ions in the local tumor microenvironment remodelling are considered a third approach for anti-tumor therapy. Therefore, we speculate about the different physical forms of magnesium that could create an anti-tumor microenvironment upon tumor interventional therapy, a technique that precisely places anti-tumor implants like particles and stents. Finally, we present our viewpoints on the potential use of magnesium in diverse solid tumor therapies to inhibit tumor progression.
Nonalcoholic fatty liver disease (NAFLD) is a multisystem metabolic disease associated with gut microflora dysbiosis and inflammation. Hydrogen (H2) is a novel and effective antiinflammatory agent. The present study was aimed to clarify the effects of 4% hydrogen (H2) inhalation on NAFLD and its mechanism of action. Sprague-Dawley rats were fed a high-fat diet for 10 weeks to induce NAFLD. Rats in treatment group inhaled 4% H2 each day for 2 h. The protective effects on hepatic histopathology, glucose tolerance, inflammatory markers, and intestinal epithelial tight junctions were assessed. Transcriptome sequencing of liver and 16 S-seq of cecal contents were also performed to explore the related mechanisms of H2 inhalation. H2 improved the hepatic histological changes and glucose tolerance, decreased the liver function parameters of plasma alanine aminotransferase and aspartate aminotransferase, and relieved liver inflammation. Liver transcriptomic data suggested that H2 treatment significantly downregulated inflammatory response genes, and the lipopolysaccharide (LPS)/Toll-like receptor (TLR) 4/nuclear transcription factor kappa B (NF-κB) signaling pathway might be involved, and the expressions of critical proteins were further validated. Meanwhile, the plasma LPS level was significantly decreased by the H2 intervention. H2 also improved the intestinal tight junction barrier by enhancing the expressions of zonula occludens-1 and occluding. Based on 16 S rRNA sequencing, H2 altered the composition of gut microbiota, improving the relative abundance of Bacteroidetes-to-Firmicutes. Collectively, our data show that H2 could prevent NAFLD induced by high-fat diet, and the anti-NAFLD effect is associated with the modulation of gut microbiota and inhibition of LPS/TLR4/NF-κB inflammatory pathway.
The growing global epidemic of obesity and type 2 diabetes mellitus has determined an increased prevalence of NAFLD (non-alcoholic fatty liver disease), making it the most common chronic liver disease in the Western world and a leading cause of liver transplantation. In the last few years, a rising number of studies conducted both on animal and human models have shown the existence of a close association between insulin resistance (IR), dysbiosis, and steatosis. However, all the mechanisms that lead to impaired permeability, inflammation, and fibrosis have not been fully clarified. Recently, new possible treatment modalities have received much attention.
To reach the review purpose, a broad-ranging literature search on multidisciplinary research databases was performed using the following terms alone or in combination: “NAFLD”, “gut dysbiosis”, “insulin resistance”, “inflammation”, “probiotics”, “Chinese herbs”.
The use of probiotics, prebiotics, symbiotics, postbiotics, fecal microbiota transplant (FMT), Chinese herbal medicine, antibiotics, diet (polyphenols and fasting diets), and minor therapies such as carbon nanoparticles, the MCJ protein, water rich in molecular hydrogen, seems to be able to improve the phenotypic pattern in NAFLD patients.
In this review, we provide an overview of how IR and dysbiosis contribute to the development and progression of NAFLD, as well as the therapeutic strategies currently in use.
In this study, earthworm (Eisenia fetida) brain was chosen as targeted receptors to probe the mechanisms of oxidative stress-related neurotoxicity, genotoxicity, and behavioral disturbances triggered by PHE. Results showed that PHE stress can initiate significant amounts of ROS, thus triggering oxidative stress in E. fetida brain. These effects were accompanied by a significant increase of damage to macromolecules DNA and lipids, resulting in severe oxidative effects. PHE exposure can induce AChE inhibition by ROS-induced injury and the accumulation of excess ACh at the nicotinic post-synaptic membrane, thus inducing aggravated neurological dysfunction and neurotoxicity of E. fetida through an oxidative stress pathway. Moreover, the burrowing behavior of earthworms was disturbed by oxidative stress-induced neurotoxicity after exposure to PHE. Furthermore, the abnormal mRNA expression profiles of oxidative stress- and neurotoxicity-related genes in worm brain were induced by PHE stress. The IBR results suggested that E. fetida brain was suffered more serious damage caused by PHE under higher doses and long-term exposure. Taken together, PHE exposure can trigger oxidative stress-mediated neurotoxicity and genotoxicity in worm brain and behavioral disorder through ROS-induced damage. This study is of great significance to evaluate the harmful effects of PHE and its mechanisms on soil ecological health.
Effects of washing crude olive pomace oil (COPO) with hydrogen-rich water (HRW) on its physicochemical and sensory properties, and phytochemicals were investigated. Acidity and peroxide values of normal water (NW) and HRW-washed COPOs decreased with a highest impact shown for HRW. The improving effects increased with the increases in washing cycle numbers. HRW-washed COPOs showed a decrease in color change and browning index. The phenolics content increased in the HRW-washed COPOs and decreased in the NW- washed COPOs. Incorporation of hydrogen into both methanol and hexane improved the extraction of phenolics (TPC), flavonoids (TFC), and antioxidants (DPPH and ABTS). Hydrogen incorporation into solvents led to a two-fold TPC and TFC increase. Washing COPO with HRW can be proposed as an effective method for improving the quality notes and preserving the nutritional value of oil, while hydrogen incorporation of solvents is recommended for better extracting phytochemicals.
As a critical component of atmospheric ultrafine particulates, ultrafine carbon black (UFCB) brings great exposure risk to organisms. At present, the action pathway and activity regulation mechanism of UFCB on functional proteins in vivo are not clear, and the size-dependent effects of UFCB during this process need to be elucidated. Superoxide dismutase (SOD), one of the most applied biomarkers to assess the environmental impact of pollutants, plays crucial roles in resistance to oxidative stress. Here, based on the inactivation of SOD (84.79 %, 86.81 % and 91.70 %) in primary mouse hepatocytes exposed to UFCB (13 nm, 50 nm and 95 nm), oxidative stress, genotoxicity and protein molecular studies were employed to elucidate the inactivation mechanisms. Results showed that inhibition of UFCB-mediated superoxide anion (O2-) contributed to a decrease in SOD activity. Furthermore, the significant increase in 8-hydroxy-2-deoxyguanosine content and the comet tail formation indicated the occurrence of DNA damage, supporting that concomitant aberrant transcriptional and protein translational under gene regulation should be responsible for SOD inactivation. At the molecular level, the constricted backbone, reduced content of α-helix and fluorescence sensitization all demonstrated that the attachment-type binding of SOD on UFCB to form the 'protein corona' disrupted protein structure. Enzyme activity assays indicated that SOD backbone tightening and helix decay resulted in decreased activity, which should be another reason for intracellular SOD inactivation. More importantly, the particle sizes of UFCB exert powerful influences on SOD inactivation mechanisms. Smaller UFCB (13 nm) induced more severe O2- inhibition and DNA damage, while UFCB50nm with the best dispersity bound more SOD and induced stronger molecular toxicity, which are their different strengths in stressing SOD inactivation in hepatocytes. Our findings provide novel insights for exploring functional proteins activity and underscore a potentially size-dependent risk of nanoparticles.
Ethnopharmacological relevance
Nonalcoholic fatty liver disease (NAFLD) is a prevalent liver disease, but currently has no specific medication in clinic. Antrodia cinnamomea (AC) is a medicinal fungus and it has been shown that AC can inhibit high fat diet (HFD)-induced lipid deposition in mouse livers, but the effective monomer in AC and mechanism against NAFLD remain unclear. It has been reported that aldehyde dehydrogenase 2 (ALDH2) activation shows protective effects on NAFLD. Our previous study demonstrates that AC and its monomer dehydroeburicoic acid (DEA) can upregulate the ALDH2 activity on alcoholic fatty liver disease mouse model, but it is not clear whether the anti-NAFLD effects of AC and DEA are mediated by ALDH2.
Aim to study
To elucidate the active compound in AC against NAFLD, study whether ALDH2 mediates the anti-NAFLD effects of AC and its effective monomer.
Materials and methods
WT mice, ALDH2−/− mice and ALDH2−/− mice re-expressed ALDH2 by lentivirus were fed with a methionine-choline deficient (MCD) diet or high fat diet (HFD) to induce NAFLD, and AC at the different doses (200 and/or 500 mg/kg body weight per day) was administrated by gavage at the same time. Primary hepatocytes derived from WT and ALDH2−/−mice were stimulated by oleic acid (OA) to induce lipid deposition, and the cells were treated with AC or DEA in the meantime. Lentivirus-mediated ALDH2-KD or ALDH2-OE were used to knock down or overexpress ALDH2 expression in HepG2 cells, respectively. Finally, the effects of DEA against NAFLD as well as its effects on upregulating liver ALDH2 and removing the harmful aldehyde 4-hydroxynonenal (4-HNE) were studied in the MCD diet-induced NAFLD mouse model.
Results
In WT mice fed with a MCD diet or HFD, AC administration reduced hepatic lipid accumulation, upregulated ALDH2 activity in mouse livers, decreased 4-HNE contents both in mouse livers and serum, inhibited lipogenesis, inflammation and oxidative stress and promoted fatty acid β-oxidation. These effects were abolished in ALDH2 KO mice but could be restored by re-expression of ALDH2 by lentivirus. In primary hepatocytes of WT mice, AC and DEA inhibited OA-induced lipid accumulation and triglyceride (TG) synthesis, promoting the β-oxidation of fatty acid in the meantime. However, these effects were lost in primary hepatocytes of ALDH2 KO mice. Moreover, the expression level of ALDH2 significantly affected the inhibitory effects of AC and DEA on OA-induced lipid deposition in HepG2 cells. The effects of AC and DEA on suppressing lipid deposition, inhibiting mitochondrial ROS levels, reducing TG synthesis, and promoting β-oxidation of fatty acid were all enhanced with the overexpression of ALDH2 and reduced with the knockdown of ALDH2 expression. DEA showed dose-dependent effects on inhibiting liver lipid deposition, elevating ALDH2 activity and reducing 4-HNE levels in the livers of MCD diet-induced NAFLD mice.
Conclusion
DEA is the effective compound in AC against NAFLD. The related anti-NAFLD mechanisms of AC and DEA were through upregulating ALDH2 expression and activity, thus enhancing the elimination of 4-HNE in the livers, and sequentially alleviating oxidative stress and inflammation, promoting fatty acid β-oxidation and decreasing lipogenesis.
Many studies have shown that redox regulation is an effective therapeutic strategy for different types of cancer. We have previously demonstrated that combined treatment with dissolved hydrogen molecule (H2) and platinum nanocolloid (Pt-nc) has carcinostatic effects and that increased intracellular reactive oxygen species (ROS) levels were closely associated with carcinostatic effects in Ehrlich mouse ascites tumor cells. However, it is unknown whether combined treatment-induced ROS generation can occur in human cancer cells. Therefore, this study aimed to examine the carcinostatic effect of the combined treatment in human cells and investigate the relationship between treatment efficacy and ROS generation. H2 and Pt-nc treatment could exert cytostatic action by inhibiting the growth of human promyelocytic leukemia HL60 and human gastric adenocarcinoma-derived NUGC-4 cells; however, no effect was observed in normal human embryo fibroblast OUMS-36 cells by the temporary exposure. These findings indicate that combined treatment with H2 and Pt-nc may act selectively in tumor cells compared with normal cells. Additionally, combined treatment with H2 and Pt-nc resulted in an approximately 200-fold increase in intracellular ROS levels compared with the control, whereas the suppressive effect of tumor cell growth was abrogated entirely by catalase treatment in NUGC-4 cells. Furthermore, combined treatment with H2 and Pt-nc induced hydrogen peroxide generation, cellular morphological changes, cell death, and a decline in DNA synthesis-positive cells. In conclusion, combined treatment with H2 and Pt-nc can induce carcinostatic/carcinocidal effects through intracellular ROS increase, morphological changes, cell death, and DNA synthesis suppression in the human tumor cell line.
Nonalcoholic steatohepatitis (NASH) could progress to hepatic fibrosis when absence of effective control. The purpose of our experiment was to investigate the protective effect of drinking water with high concentration of hydrogen in our study named hydrogen rich water (HRW) on mice with non-alcoholic fatty liver disease to elucidate the mechanism underlying the molecular hydrogen therapeutic action. The choline-supplemented, L-amino acid-defined (CSAA) or choline-deficient, L-amino acid-defined (CDAA) diet for 20 weeks were used to induce NASH and fibrosis in the mice model and simultaneously treated with HRW for different periods of time. Primary hepatocytes were stimulated by palmitate in order to mimic a liver lipid metabolism during fatty liver formation. Mice in the CSAA + HRW group had lower serum levels of ALT and AST and milder histological damage. The inflammatory cytokines were expressed at lower levels in the HRW group than in the CSAA group. Importantly, HRW reversed hepatocyte apoptosis as well as hepatic inflammation and fibrosis in pre-existing hepatic fibrosis specimens. Molecular hydrogen inhibits the lipopolysaccharide-induced production of inflammation cytokines through an HO-1/IL-10-independent pathway. Furthermore, HRW improved hepatic steatosis in the CSAA + HRW group. Sirt1 induction by molecular hydrogen via the HO-1/AMPK/PPARα/PPARγ pathway suppresses palmitate-mediated abnormal fat metabolism. Orally administered HRW suppressed steatosis induced by CSAA and attenuated fibrosis induced by CDAA, possibly by reducing oxidative stress and the inflammation response.
Aims
Oxidized phospholipids (OxPLs) are formed as a result of oxidative stress, which potentially mediate multiple pathological effects. We aimed to evaluate the effects of hydrogen (H2) on OxPLs in vivo and the underlying mechanism.
Main methods
Rats were randomly assigned to three groups: control group fed with a chow diet, model group fed with a high-fat diet, and H2-treated group fed with a high-fat diet and treated by 4% H2 inhalation for ten weeks. OxPLs in liver and plasma were analyzed by liquid chromatography–mass spectrometry. High-density lipoprotein (HDL) was separated by ultracentrifugation. A proteomic analysis was performed to reveal the alternation of HDL protein composition and he antioxidant capacity of HDL was tested by low-density lipoprotein oxidation experiment. Furthermore, the activity or expression of HDL-associated enzymes were evaluated.
Key findings
Inhalation of 4% H2 decreased the accumulation of OxPLs in rats. In vitro tests revealed that the different concentrations of H2 did not inhibit the formation of OxPLs mediated by non-enzymatic oxidation. H2 inhalation altered the components and enhanced the anti-oxidative capacity of HDL in rats fed with a high-fat diet. Further experiments showed that H2 significantly regulated the activity of lipoprotein-associated phospholipase A2, paraoxonase-1, and the expression of lecithin:cholesterol acyltransferase.
Significance
Our findings revealed that H2 may reduce the OxPLs levels through its influence on HDL-associated enzymes that can act on OxPLs, suggesting that H2 can be used in alleviating diseases related to lipid peroxidation due to oxidative stress.
Water‐splitting has been extensively studied especially for energy applications. It is often not paid with enough attention for biomedical applications. In fact, several innovative breakthroughs have been achieved in the past few years by employing water‐splitting for treating cancer and other diseases. Interestingly, among these important works, only two reports have mentioned the term “water‐splitting.” For this reason, the importance of water‐splitting for biomedical applications is significantly underestimated. This progress work is written with the aims to explain and summarize how the principle of water‐splitting is employed to achieve therapeutic results not offered by conventional approaches. It is expected that this progress report will not only explain the importance of water‐splitting to scientists in the biomedical fields, it should also draw attention from scientists working on energy applications of water‐splitting.
The use of hydrogen for cancer control has made great progress in cytology and animal experiments. With the increasing number of hydrogen products on the market, larger numbers of advanced cancer patients have participated in clinical trials or received treatment at home after purchase. Our study reported a real-world survey from 82 patients with good cancer control using hydrogen products, including real world evidence from patients who received ineffective traditional treatment, patients who received traditional treatment that failed, or patients who refused traditional treatment. Two typical cases were reported herein. Subsequently, we included studies on the mechanism of hydrogen oncology. The mechanism of cancer control using hydrogen includes the inhibition of tumor cells and the activation of exhausted lymphocytes. Large-scale real world evidence has shown clinical value, and yet remains to be further developed and researched.
Molecular hydrogen (H2 ) was long regarded as non-functional in mammalian cells. We overturned the concept by demonstrating that H2 exhibits antioxidant effects and protects cells against oxidative stress. Subsequently, it has been revealed that H2 has multiple functions in addition to antioxidant effects, including ant-inflammatory, anti-allergic functions, and as a cell death and autophagy regulation. Additionally, H2 stimulates energy metabolism. Because H2 does not readily react with most biomolecules without a catalyst, it is essential to identify the primary targets with which H2 reacts or interacts directly. As a first event, H2 may react directly with strong oxidants such as hydroxyl radicals (•OH) in vivo. This review addresses the key issues related to this in vivo reaction. •OH may have a physiological role because it triggers a free radical chain reaction and may be involved in the regulation of Ca2+ - or mitochondrial ATP-dependent K+ - channeling. In the subsequent pathway, H2 suppressed a free radical chain reaction, leading to decreases in lipid peroxide and its end products. Derived from the peroxides, 4-hydroxy-2-nonenal functions as a mediator that up-regulates multiple functional PGC-1α. As the other direct target in vitro and in vivo, H2 intervenes in the free radical chain reaction to modify oxidized phospholipids, which may act as an antagonist of Ca2+ -channels. The resulting suppression of Ca2+ - signaling inactivates multiple functional NFAT and CREB transcription factors, which may explain H2 multifunctionality. This review also addresses the involvement of NFAT in the beneficial role of H2 in COVID-19, Alzheimer’s disease and advanced cancer. We discuss some unsolved issues of H2 action on lipopolysaccharide signaling, MAPK and NF-κB pathways and the Nrf2 paradox. Finally, as a novel idea for the direct targeting of H2 , this review introduces the possibility that H2 causes structural changes in proteins via hydrate water changes.
Obesity, a major health concern, results from an imbalance between energy intake and expenditure. Leptin-deficient ob/ob mice are paradigmatic of obesity, resulting from excess energy intake and storage. Mice lacking acyl-CoA oxidase 1 (Acox1), the first enzyme of the peroxisomal fatty acid β-oxidation system, are characterized by increased energy expenditure and a lean body phenotype caused by sustained activation of peroxisome proliferator-activated receptor α (PPARα) by endogenous ligands in liver that remain unmetabolized in the absence of Acox1. We generated ob/ob mice deficient in Acox1 (Acox1(-/-)) to determine how the activation of PPARα by endogenous ligands might affect the obesity of ob/ob mice. In contrast to Acox1(-/-) (14.3±1.2 g at 6 mo) and the Acox1-deficient (ob/ob) double-mutant mice (23.8±4.6 g at 6 mo), the ob/ob mice are severely obese (54.3±3.2 g at 6 mo) and had significantly more (P<0.01) epididymal fat content. The resistance of Acox1(-/-)/ob/ob mice to obesity is due to increased PPARα-mediated up-regulation of genes involved in fatty acid oxidation in liver. Activation of PPARα in Acox1-deficient ob/ob mice also reduces serum glucose and insulin (P<0.05) and improves glucose tolerance and insulin sensitivity. Further, PPARα activation reduces hepatic steatosis and increases hepatocellular regenerative response in Acox1(-/-)/ob/ob mice at a more accelerated pace than in mice lacking only Acox1. However, Acox1(-/-)/ob/ob mice manifest hepatic endoplasmic reticulum (ER) stress and also develop hepatocellular carcinomas (8 of 8 mice) similar to those observed in Acox1(-/-) mice (10 of 10 mice), but unlike in ob/ob (0 of 14 mice) and OB/OB (0 of 6 mice) mice, suggesting that superimposed ER stress and PPARα activation contribute to carcinogenesis in a fatty liver. Finally, absence of Acox1 in ob/ob mice can impart resistance to high-fat diet (60% fat)-induced obesity, and their liver had significantly (P<0.01) more cell proliferation. These studies with Acox1(-/-)/ob/ob mice indicate that sustained activation of lipid-sensing nuclear receptor PPARα attenuates obesity and restores glucose homeostasis by ameliorating insulin resistance but increases the risk for liver cancer development, in part related to excess energy combustion.
C57BL/6J and 129S6/Sv (B6 and 129) mice differ dramatically in their susceptibility to developing diabetes in response to diet- or genetically induced insulin resistance. A major locus contributing to this difference has been mapped to a region on mouse chromosome 14 that contains the gene encoding PKCδ. Here, we found that PKCδ expression in liver was 2-fold higher in B6 versus 129 mice from birth and was further increased in B6 but not 129 mice in response to a high-fat diet. PRKCD gene expression was also elevated in obese humans and was positively correlated with fasting glucose and circulating triglycerides. Mice with global or liver-specific inactivation of the Prkcd gene displayed increased hepatic insulin signaling and reduced expression of gluconeogenic and lipogenic enzymes. This resulted in increased insulin-induced suppression of hepatic gluconeogenesis, improved glucose tolerance, and reduced hepatosteatosis with aging. Conversely, mice with liver-specific overexpression of PKCδ developed hepatic insulin resistance characterized by decreased insulin signaling, enhanced lipogenic gene expression, and hepatosteatosis. Therefore, changes in the expression and regulation of PKCδ between strains of mice and in obese humans play an important role in the genetic risk of hepatic insulin resistance, glucose intolerance, and hepatosteatosis; and thus PKCδ may be a potential target in the treatment of metabolic syndrome.
While type 2 diabetes is an independent risk factor for worsening of human non-alcoholic steatohepatitis (NASH) in clinical studies, it has not been systematically reported in any model whether diabetes exacerbates NASH. The study aim was to determine if diabetes causes NASH progression in a mouse model of diet induced obesity.
C57BL/6 mice were fed a high fat diet (HFD: 45% kcal fat) or standard chow (CHOW: 12% kcal fat) for 20 weeks and some animals (HFD+DM or CHOW+DM) were also rendered diabetic by low dose streptozotocin for the final 5 weeks, to model type 2 diabetes. Serum assays included circulating insulin, triglyceride, ALT and AST, glucose, and ultrasensitive CRP and results of insulin tolerance tests. Intrahepatic lipid, triglyceride, macrophage infiltration, and fibrosis were determined. Fibrosis markers collagen-I, collagen-III, CTGF, TIMP-1, and FAP were assessed by qPCR and CTGF and collagen-I by immunostaining.
HFD mice were obese, insulin resistant and hyperinsulinaemic, with NASH features of elevated intrahepatic lipid and macrophages, but without fibrosis. In contrast, the HFD+DM mice exhibited fibrosis in addition to these NASH features. By ANOVA, Sirius red staining at perisinusoidal, portal tract and central vein sites, collagen-I, collagen-III, FAP, and TIMP-1 transcripts and collagen-I and CTGF protein were each significantly increased in HFD+DM, compared with CHOW alone. In a further experiment, insulin treatment protected against fibrosis and CRP increases in HFD+DM, showing that diabetes, not streptozotocin, causes the fibrosis.
This novel model indicates that diet-induced NASH fibrosis is exacerbated by diabetes and attenuated by insulin therapy.
Male Sprague-Dawley rats were chronically fed a high-unsaturated-fat diet for 130 days by using total enteral nutrition (TEN), or the same diet in which ethanol (EtOH) isocalorically replaced carbohydrate calories. Additional groups were supplemented with the antioxidant N-acetylcysteine (NAC) at 1.7 g·kg(-1)·day(-1). Relative to an ad libitum chow-fed group, the high-fat-fed controls had three- to fourfold greater expression of fatty acid transporter CD36 mRNA and developed mild steatosis but little other hepatic pathology. NAC treatment resulted in increased somatic growth relative to controls (4.0 ± 0.1 vs. 3.1 ± 0.1 g/day) and increased hepatic steatosis score (3.5 ± 0.6 vs. 2.7 ± 1.2), associated with suppression of the triglyceride hydrolyzing protein adiponutrin, but produced no elevation in serum alanine aminotransferase (ALT). Chronic EtOH treatment increased expression of fatty acid transport protein FATP-2 mRNA twofold, resulting in marked hepatic steatosis, oxidative stress, and a twofold elevation in serum ALT. However, no changes in tumor necrosis factor-α or transforming growth factor-β expression were observed. Fibrosis, as measured by Masson's trichrome and picrosirius red staining, and a twofold increase in expression of type I and type III collagen mRNA, was only observed after EtOH treatment. Long-term EtOH treatment increased hepatocyte proliferation but did not modify the hepatic mRNAs for hedgehog pathway ligands or target genes or genes regulating epithelial-to-mesenchymal transition. Although the effects of NAC on EtOH-induced fibrosis could not be fully evaluated, NAC had additive effects on hepatocyte proliferation and prevented EtOH-induced oxidative stress and necrosis, despite a failure to reverse hepatic steatosis.
Periodontitis induces excessive production of reactive oxygen species in periodontal lesions. This may impair circulating pro-oxidant/anti-oxidant balance and induce the oxidation of low-density lipoprotein (LDL) in blood. The purpose of this study was to monitor circulating oxidized LDL and oxidative stress in subjects with chronic periodontitis following non-surgical periodontal treatment. Plasma levels of oxidized LDL and oxidative stress in 22 otherwise healthy non-smokers with chronic periodontitis (mean age 44.0 years) were measured at baseline and at 1 and 2 months after non-surgical periodontal treatment. At baseline, chronic periodontitis patients had higher plasma levels of oxidized LDL and oxidative stress than healthy subjects (p < 0.001). Periodontal treatment was associated with a significant reduction in plasma levels of oxidized LDL (oxLDL)(p < 0.001) and oxidative stress (p < 0.001). At 2 months after periodontal treatment, the degree of change in the oxLDL was positively correlated with that in the oxidative stress (r = 0.593, p = 0.004). These observations indicate that periodontitis patients showed higher levels of circulating oxLDL and oxidative stress than healthy subjects. In addition, improved oral hygiene and non-surgical periodontal treatment were effective in decreasing oxLDL, which was positively associated with a reduction in circulating oxidative stress.
Nonalcoholic steatohepatitis is a common liver disease that can progress to cirrhosis. Currently, there is no established treatment for this disease.
We randomly assigned 247 adults with nonalcoholic steatohepatitis and without diabetes to receive pioglitazone at a dose of 30 mg daily (80 subjects), vitamin E at a dose of 800 IU daily (84 subjects), or placebo (83 subjects), for 96 weeks. The primary outcome was an improvement in histologic features of nonalcoholic steatohepatitis, as assessed with the use of a composite of standardized scores for steatosis, lobular inflammation, hepatocellular ballooning, and fibrosis. Given the two planned primary comparisons, P values of less than 0.025 were considered to indicate statistical significance.
Vitamin E therapy, as compared with placebo, was associated with a significantly higher rate of improvement in nonalcoholic steatohepatitis (43% vs. 19%, P=0.001), but the difference in the rate of improvement with pioglitazone as compared with placebo was not significant (34% and 19%, respectively; P=0.04). Serum alanine and aspartate aminotransferase levels were reduced with vitamin E and with pioglitazone, as compared with placebo (P<0.001 for both comparisons), and both agents were associated with reductions in hepatic steatosis (P=0.005 for vitamin E and P<0.001 for pioglitazone) and lobular inflammation (P=0.02 for vitamin E and P=0.004 for pioglitazone) but not with improvement in fibrosis scores (P=0.24 for vitamin E and P=0.12 for pioglitazone). Subjects who received pioglitazone gained more weight than did those who received vitamin E or placebo; the rates of other side effects were similar among the three groups.
Vitamin E was superior to placebo for the treatment of nonalcoholic steatohepatitis in adults without diabetes. There was no benefit of pioglitazone over placebo for the primary outcome; however, significant benefits of pioglitazone were observed for some of the secondary outcomes. (ClinicalTrials.gov number, NCT00063622.)
Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological term that encompasses a spectrum of abnormalities ranging from simple triglyceride accumulation in the hepatocytes (hepatic steatosis) to hepatic steatosis with inflammation (steatohepatitis, also known as nonalcoholic steatohepatitis or NASH). NASH can also progress to cirrhosis and hepatocellular carcinoma (HCC). Steatohepatitis has been estimated to affect around 5% of the total population and 20% of those who are overweight. The mechanisms leading to NASH and its progression to cirrhosis and HCC remain unclear, but it is a condition typically associated with obesity, insulin resistance, diabetes, and hypertriglyceridemia. This point corroborates the need for animal models and molecular markers that allow us to understand the mechanisms underlying this disease. Nowadays, there are numerous mice models to study abnormal liver function such as steatosis, NASH, and hepatocellular carcinoma. The study of the established animal models has provided many clues in the pathogenesis of steatosis and steatohepatitis, although these remain incompletely understood and no mice model completely fulfills the clinical features observed in humans.
In addition, there is a lack of accurate sensitive diagnostic tests that do not involve invasive procedures. Current laboratory tests include some biochemical analysis, but their utility for diagnosing NASH is still poor. For that reason, a great effort is being made toward the identification and validation of novel biomarkers to assess NASH using high-throughput analysis based on genomics, proteomics, and metabolomics. The most recent discoveries and their validation will be discussed.
Small intestinal bacterial overgrowth may contribute to the development of non-alcoholic steatohepatitis, perhaps by increasing intestinal permeability and promoting the absorption of endotoxin or other enteric bacterial products.
To investigate the prevalence of small intestinal bacterial overgrowth, increased intestinal permeability, elevated endotoxin, and tumour necrosis factor alpha (TNF-alpha) levels in patients with non-alcoholic steatohepatitis and in control subjects.
Twenty two patients with non-alcoholic steatohepatitis and 23 control subjects were studied. Small intestinal bacterial overgrowth was assessed by a combined (14)C-D-xylose and lactulose breath test. Intestinal permeability was assessed by a dual lactulose-rhamnose sugar test. Serum endotoxin levels were determined using the limulus amoebocyte lysate assay and TNF-alpha levels using an ELISA.
Small intestinal bacterial overgrowth was present in 50% of patients with non-alcoholic steatosis and 22% of control subjects (p=0.048). Mean TNF-alpha levels in non-alcoholic steatohepatitis patients and control subjects were 14.2 and 7.5 pg/ml, respectively (p=0.001). Intestinal permeability and serum endotoxin levels were similar in the two groups.
Patients with non-alcoholic steatohepatitis have a higher prevalence of small intestinal bacterial overgrowth, as assessed by the (14)C-D-xylose-lactulose breath test, and higher TNF-alpha levels in comparison with control subjects. This is not accompanied by increased intestinal permeability or elevated endotoxin levels.
Acute oxidative stress induced by ischemia-reperfusion or inflammation causes serious damage to tissues, and persistent oxidative stress is accepted as one of the causes of many common diseases including cancer. We show here that hydrogen (H(2)) has potential as an antioxidant in preventive and therapeutic applications. We induced acute oxidative stress in cultured cells by three independent methods. H(2) selectively reduced the hydroxyl radical, the most cytotoxic of reactive oxygen species (ROS), and effectively protected cells; however, H(2) did not react with other ROS, which possess physiological roles. We used an acute rat model in which oxidative stress damage was induced in the brain by focal ischemia and reperfusion. The inhalation of H(2) gas markedly suppressed brain injury by buffering the effects of oxidative stress. Thus H(2) can be used as an effective antioxidant therapy; owing to its ability to rapidly diffuse across membranes, it can reach and react with cytotoxic ROS and thus protect against oxidative damage.
The methionine choline-deficient (MCD) diet results in liver injury similar to human nonalcoholic steatohepatitis (NASH). The aims of this study were to define mechanisms of MCD-induced steatosis in insulin-resistant db/db and insulin-sensitive db/m mice. MCD-fed db/db mice developed more hepatic steatosis and retained more insulin resistance than MCD-fed db/m mice. Both subcutaneous and gonadal fat were reduced by MCD feeding: gonadal fat decreased by 23% in db/db mice and by 90% in db/m mice. Weight loss was attenuated in the db/db mice, being only 13% compared with 35% in MCD-fed db/db and db/m mice, respectively. Both strains had upregulation of hepatic fatty acid transport proteins as well as increased hepatic uptake of [14C]oleic acid: 3-fold in db/m mice (P < 0.001) and 2-fold in db/db mice (P < 0.01) after 4 weeks of MCD feeding. In both murine strains, the MCD diet reduced triglyceride secretion and downregulated genes involved in triglyceride synthesis. Therefore, increased fatty acid uptake and decreased VLDL secretion represent two important mechanisms by which the MCD diet promotes intrahepatic lipid accumulation in this model. Feeding the MCD diet to diabetic rodents broadens the applicability of this model for the study of human NASH.
Reactive oxygen species (ROS) are considered to play a prominent causative role in the development of various hepatic disorders. Antioxidants have been effectively demonstrated to protect against hepatic damage. Hydrogen (H(2)), a new antioxidant, was reported to selectively reduce the strongest oxidants, such as hydroxyl radicals (·OH) and peroxynitrite (ONOO(-)), without disturbing metabolic oxidation-reduction reactions or disrupting ROS involved in cell signaling. In place of H(2) gas, hydrogen-rich saline (HS) may be more suitable for clinical application. We herein aim to verify its protective effects in experimental models of liver injury.
H(2) concentration in vivo was detected by hydrogen microelectrode for the first time. Liver damage, ROS accumulation, cytokine levels, and apoptotic protein expression were, respectively, evaluated after GalN/LPS, CCl(4), and DEN challenge. Simultaneously, CCl(4)-induced hepatic cirrhosis and DEN-induced hepatocyte proliferation were measured.
HS significantly increased hydrogen concentration in liver and kidney tissues. As a result, acute liver injury, hepatic cirrhosis, and hepatocyte proliferation were reduced through the quenching of detrimental ROS. Activity of pro-apoptotic players, such as JNK and caspase-3, were also inhibited.
HS could protect against liver injury and also inhibit the processes leading to liver cirrhosis and hepatocyte compensatory proliferation.
Mitochondria are the major source of oxidative stress. Acute oxidative stress causes serious damage to tissues, and persistent oxidative stress is one of the causes of many common diseases, cancer and the aging process; however, there has been little success in developing an effective antioxidant with no side effect. We have reported that molecular hydrogen has potential as an effective antioxidant for medical applications [Ohsawa et al., Nat. Med. 13 (2007) 688-694].
We review the recent progress toward therapeutic and preventive applications of hydrogen. Since we published the first paper in Nature Medicine, effects of hydrogen have been reported in more than 38 diseases, physiological states and clinical tests in leading biological/medical journals. Based on this cumulative knowledge, the beneficial biological effects of hydrogen have been confirmed. There are several ways to intake or consume hydrogen, including inhaling hydrogen gas, drinking hydrogen-dissolved water, taking a hydrogen bath, injecting hydrogen-dissolved saline, dropping hydrogen-dissolved saline into the eyes, and increasing the production of intestinal hydrogen by bacteria. Hydrogen has many advantages for therapeutic and preventive applications, and shows not only anti-oxidative stress effects, but also has various anti-inflammatory and anti-allergic effects. Preliminary clinical trials show that drinking hydrogen-dissolved water seems to improve the pathology of mitochondrial disorders.
Hydrogen has biological benefits toward preventive and therapeutic applications; however, the molecular mechanisms underlying the marked effects of small amounts of hydrogen remain elusive.
Hydrogen is a novel antioxidant with great potential for actual medical applications. This article is part of a Special Issue entitled Biochemistry of Mitochondria.
The effects of the administration of molecular hydrogen-saturated drinking water (hydrogen water) on hepatic gene expression were investigated in rats. Using DNA microarrays, 548 upregulated and 695 downregulated genes were detected in the liver after 4 weeks of administration of hydrogen water. Gene Ontology analysis revealed that genes for oxidoreduction-related proteins, including hydroxymethylglutaryl CoA reductase, were significantly enriched in the upregulated genes.
The pathogenesis of nonalcoholic steatohepatitis (NASH) and inflammasome activation involves sequential hits. The inflammasome, which cleaves pro-interleukin-1beta (pro-IL-1beta) into secreted IL-1beta, is induced by endogenous and exogenous danger signals. Lipopolysaccharide (LPS), a toll-like receptor 4 ligand, plays a role in NASH and also activates the inflammasome. In this study, we hypothesized that the inflammasome is activated in NASH by multiple hits involving endogenous and exogenous danger signals.
METHODS: Using mouse models of methionine choline-deficient (MCD) diet-induced NASH and high-fat diet-induced NASH, we found up-regulation of the inflammasome [including NACHT, LRR, and PYD domains-containing protein 3 (NALP3; cryopyrin), apoptosis-associated speck-like CARD-domain containing protein, pannexin-1, and pro-caspase-1] at the messenger RNA (mRNA) level increased caspase-1 activity, and mature IL-1beta protein levels in mice with steatohepatitis in comparison with control livers. There was no inflammasome activation in mice with only steatosis. The MCD diet sensitized mice to LPS-induced increases in NALP3, pannexin-1, IL-1beta mRNA, and mature IL-1beta protein levels in the liver. We demonstrate for the first time that inflammasome activation occurs in isolated hepatocytes in steatohepatitis. Our novel data show that the saturated fatty acid (FA) palmitic acid (PA) activates the inflammasome and induces sensitization to LPS-induced IL-1beta release in hepatocytes. Furthermore, PA triggers the release of danger signals from hepatocytes in a caspase-dependent manner. These hepatocyte-derived danger signals, in turn, activate inflammasome, IL-1beta, and tumor necrosis factor alpha release in liver mononuclear cells.
CONCLUSION: Our novel findings indicate that saturated FAs represent an endogenous danger in the form of a first hit, up-regulate the inflammasome in NASH, and induce sensitization to a second hit with LPS for IL-beta release in hepatocytes. Furthermore, hepatocytes exposed to saturated FAs release danger signals that trigger inflammasome activation in immune cells. Thus, hepatocytes play a key role in orchestrating tissue responses to danger signals in NASH.
Recent extensive studies have revealed that molecular hydrogen (H(2)) has great potential for improving oxidative stress-related diseases by inhaling H(2) gas, injecting saline with dissolved H(2), or drinking water with dissolved H(2) (H(2)-water); however, little is known about the dynamic movement of H(2) in a body. First, we show that hepatic glycogen accumulates H(2) after oral administration of H(2)-water, explaining why consumption of even a small amount of H(2) over a short span time efficiently improves various disease models. This finding was supported by an in vitro experiment in which glycogen solution maintained H(2). Next, we examined the benefit of ad libitum drinking H(2)-water to type 2 diabetes using db/db obesity model mice lacking the functional leptin receptor. Drinking H(2)-water reduced hepatic oxidative stress, and significantly alleviated fatty liver in db/db mice as well as high fat-diet-induced fatty liver in wild-type mice. Long-term drinking H(2)-water significantly controlled fat and body weights, despite no increase in consumption of diet and water. Moreover, drinking H(2)-water decreased levels of plasma glucose, insulin, and triglyceride, the effect of which on hyperglycemia was similar to diet restriction. To examine how drinking H(2)-water improves obesity and metabolic parameters at the molecular level, we examined gene-expression profiles, and found enhanced expression of a hepatic hormone, fibroblast growth factor 21 (FGF21), which functions to enhance fatty acid and glucose expenditure. Indeed, H(2) stimulated energy metabolism as measured by oxygen consumption. The present results suggest the potential benefit of H(2) in improving obesity, diabetes, and metabolic syndrome.
Liver fibrogenesis is sustained by myofibroblast-like cells originating from hepatic stellate cells (HSC/MFs), portal fibroblasts or bone marrow-derived cells, including mesenchymal stem cells (MSCs). Herein, we investigated the mechanistic role of intracellular generation of reactive oxygen species (ROS) and redox-sensitive signal transduction pathways in mediating chemotaxis, a critical profibrogenic response for human HSC/MFs and for MSC potentially engrafting chronically injured liver.
Intracellular generation of ROS and signal transduction pathways were evaluated by integrating morphological and molecular biology techniques. Chemokinesis and chemotaxis were evaluated by wound healing assay and modified Boyden's chamber assay, respectively. Additional in vivo evidence was obtained in human specimens from HCV-related cirrhosis.
Human MSCs and HSC/MFs migrate in response to a panel of polypeptide chemoattractants and extracellularly generated superoxide anion. All polypeptides induced a NADPH-oxidase-dependent intracellular rise in ROS, resulting in activation of ERK1/2 and JNK1/2. Moreover, menadione or 2,3-dimethoxy-1,4-naphthoquinone, which generate intracellular superoxide anion or hydrogen peroxide, respectively, induced ERK1/2 and JNK1/2 activation and migration. JNK1 activation was predominant for migration as shown by specific silencing. Finally, activation of ERK1/2 and JNK1/2 was found in extracts obtained from HSC/MFs during the course of an oxidative stress-mediated model of liver injury and phosphorylated JNK1/2 isoforms were detected in α-smooth muscle actin-positive myofibroblasts lining fibrotic septa in human cirrhotic livers.
Intracellular generation of ROS, through activation of specific signaling pathways, is a critical event for directional migration of HSC/MFs and MSCs.
Unlabelled:
Nonalcoholic fatty liver disease (NAFLD) encompasses a histological spectrum ranging from simple steatosis to nonalcoholic steatohepatitis (NASH). NAFLD carries a higher risk of cardio-metabolic and liver-related complications, the latter being confined to NASH and demanding specific treatment. We assessed the efficacy of proposed treatments for NAFLD/NASH by reviewing reports of randomized controlled trials (RCTs) on online databases and national and international meeting abstracts through January 2010. Primary outcome measure was histological improvement; secondary outcome was biochemical improvement; improvement in radiological steatosis was also evaluated. Two reviewers extracted articles using predefined quality indicators, independently and in duplicate. Main outcomes of randomized controlled trials (RCTs) were pooled using random-effects or fixed-effects models. Publication bias was assessed by funnel plots. Forty-nine RCTs (30 in NASH) were included: 23 RCTs (22 in NASH, 1 in NAFLD) had post-treatment histology. Most RCTs were small and did not exceed 1-year duration. Weight loss, thiazolidinediones (especially pioglitazone), and antioxidants were most extensively evaluated. Weight loss was safe and dose-dependently improved histological disease activity in NASH, but more than 50% of patients failed to achieve target weight loss. Thiazolidinediones improved steatosis and inflammation but yielded significant weight gain. RCTs with antioxidants yielded conflicting results and were heterogeneous with respect to type and dose of drug, duration, implementation of lifestyle intervention. Among the other agents, pentoxifylline, telmisartan and L-carnitine improved liver histology in at least 1 RCT in NASH; polyunsaturated fatty acid (PUFA) ameliorated biochemical and radiological markers of NAFLD. Other approaches yielded negative results.
Conclusion:
Well-designed RCTs of adequate size and duration, with histological endpoints, are needed to assess long-term safety and efficacy of proposed treatments on patient-oriented clinical outcomes.
Reactive oxygen species (ROS) contribute to the development of interstitial fibrosis and tubular atrophy seen in chronic allograft nephropathy (CAN). As molecular hydrogen gas can act as a scavenger of ROS, we tested the effect of treatment with hydrogen water (HW) in a model of kidney transplantation, in which allografts from Lewis rats were orthotopically transplanted into Brown Norway recipients that had undergone bilateral nephrectomy. Molecular hydrogen was dissolved in water and recipients were given HW from day 0 until day 150. Rats that were treated with regular water (RW) gradually developed proteinuria and their creatinine clearance declined, ultimately leading to graft failure secondary to CAN. In contrast, treatment with HW improved allograft function, slowed the progression of CAN, reduced oxidant injury and inflammatory mediator production, and improved overall survival. Inflammatory signaling pathways, such as mitogen-activated protein kinases, were less activated in renal allografts from HW-treated rats as compared with RW-treated rats. Hence, oral HW is an effective antioxidant and antiinflammatory agent that prevented CAN, improved survival of rat renal allografts, and may be of therapeutic value in the setting of transplantation.
Aims: Hepatic steatosis and iron cause oxidative stress, thereby progressing steatosis to steatohepatitis. We quantified the expression of genes involved in the metabolism of fatty acids and iron in patients with nonalcoholic fatty liver disease (NAFLD).
Methods: The levels of transcripts for the following genes were quantified from biopsy specimens of 74 patients with NAFLD: thioredoxin (Trx), fatty acid transport protein 5 (FATP5), sterol regulatory element-binding protein 1c (SREBP1c), fatty acid synthase (FASN), acetyl-coenzyme A carboxylase (ACAC), peroxisome proliferative activated receptor α (PPARα), cytochrome P-450 2E1 (CYP2E1), acyl-coenzyme A dehydrogenase (ACADM), acyl-coenzyme A oxidase (ACOX), microsomal triglyceride transfer protein (MTP), transferrin receptor 1 (TfR1), transferrin receptor 2 (TfR2) and hepcidin. Twelve samples of human liver RNA were used as controls. Histological evaluation followed the methods of Brunt.
Results: The levels of all genes were significantly higher in the NAFLD patients than in controls. The Trx level increased as the stage progressed. The levels of FATP5, SREBP1c, ACAC, PPARα, CYP2E1, ACADM and MTP significantly decreased as the stage and grade progressed (P < 0.05). Hepatic iron score (HIS) increased as the stage progressed. The TfR1 level significantly increased as the stage progressed (P < 0.05), whereas TfR2 level significantly decreased (P < 0.05). The ratio of hepcidin mRNA/ferritin (P < 0.001) or hepcidin mRNA/HIS (P < 0.01) was significantly lower in NASH patients than simple steatosis patients.
Conclusions: Steatosis-related metabolism is attenuated as NAFLD progresses, whereas iron-related metabolism is exacerbated. Appropriate therapies should be considered on the basis of metabolic changes.
Ethnic differences in non-alcoholic steatohepatitis (NASH) are well-documented, but there has been no study on the prognosis of Japanese NASH patients with cirrhosis. Accordingly, we compared cirrhotic NASH with liver cirrhosis caused by chronic hepatitis C (LC-C) to clarify its clinical features and define the risk factors for death.
A prospective evaluation of the outcomes of NASH patients with severe fibrosis was started in 1990. Data on age- and sex-matched patients with biopsy-proven LC-C were collected retrospectively and used as the control.
There were 68 patients with cirrhotic NASH and 69 with LC-C. The Child-Turcotte-Pugh (CTP) class was similar in these two groups. Although the outcome of the NASH group was better than that of the LC-C group, cirrhotic NASH followed a similar course to that of LC-C; that is, complications of cirrhosis developed, including hepatocellular carcinoma (HCC; the 5-year HCC rate was 11.3% for NASH and 30.5% for HCV) and death (the 5-year survival rates were 75.2% and 73.8%, respectively). HCC was the leading cause of death in both groups (NASH, 47%; HCV, 68%). The occurrence of HCC and the CTP class were significant risk factors for mortality in NASH patients according to a multivariate analysis (HCC: hazard ratio [HR] 7.96, 95% confidence interval [CI] 2.45-25.88, CTP class A: HR 0.17, 95% CI 0.06-0.50).
In conclusion, the present study confirmed that cirrhotic NASH has a similar course to LC-C. The occurrence of HCC was the strongest predictor of mortality in the NASH groups. These findings may be helpful when deciding on therapeutic interventions for NASH and also for the daily management of these patients.
Development of hepatocellular carcinomas in rats caused by a choline-deficient, L-amino acid-defined (CDAA) diet, usually associated with fatty liver, fibrosis, cirrhosis and oxidative DNA damage, has been recognized as a useful model of hepatocarcinogenesis caused by endogenous factors. In the present study, in order to further explore involved factors and genes, we established an equivalent model in spontaneous liver tumor-resistant C57BL/6J mice. Six-week-old males and females were continuously fed the CDAA diet and histological liver lesions and oxidative DNA damage due to 8-hydroxydeoxyguanosine (8-OHdG) were examined after 22, 65 and 84 weeks. In male mice, fatty change and fibrosis were evident at 22 weeks, and preneoplastic foci of altered hepatocytes were seen at an incidence of 8/8 (100%) and a multiplicity of 6.6 +/- 4.0 per mouse at 65 weeks. Hepatocellular adenomas and carcinomas developed at incidences of 16/24 (66.7%) and 5/24 (20.8%), and multiplicities of 1.42 +/- 1.32 and 0.29 +/- 0.62, respectively, at 84 weeks. The female mice exhibited resistance to development of these lesions. The CDAA diet also increased 8-OHdG levels in male but not female mice. These results indicate that a CDAA diet causes hepatocellular preneoplastic foci, adenomas and carcinomas associated with fibrosis and oxidative DNA damage in mice, as in rats, providing a hepatocarcinogenesis model caused by endogenous factors in mice.
Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic steatosis in the absence of a history of significant alcohol use or other known liver disease. Nonalcoholic steatohepatitis (NASH) is the progressive form of NAFLD. The Pathology Committee of the NASH Clinical Research Network designed and validated a histological feature scoring system that addresses the full spectrum of lesions of NAFLD and proposed a NAFLD activity score (NAS) for use in clinical trials. The scoring system comprised 14 histological features, 4 of which were evaluated semi-quantitatively: steatosis (0-3), lobular inflammation (0-2), hepatocellular ballooning (0-2), and fibrosis (0-4). Another nine features were recorded as present or absent. An anonymized study set of 50 cases (32 from adult hepatology services, 18 from pediatric hepatology services) was assembled, coded, and circulated. For the validation study, agreement on scoring and a diagnostic categorization ("NASH," "borderline," or "not NASH") were evaluated by using weighted kappa statistics. Inter-rater agreement on adult cases was: 0.84 for fibrosis, 0.79 for steatosis, 0.56 for injury, and 0.45 for lobular inflammation. Agreement on diagnostic category was 0.61. Using multiple logistic regression, five features were independently associated with the diagnosis of NASH in adult biopsies: steatosis (P = .009), hepatocellular ballooning (P = .0001), lobular inflammation (P = .0001), fibrosis (P = .0001), and the absence of lipogranulomas (P = .001). The proposed NAS is the unweighted sum of steatosis, lobular inflammation, and hepatocellular ballooning scores. In conclusion, we present a strong scoring system and NAS for NAFLD and NASH with reasonable inter-rater reproducibility that should be useful for studies of both adults and children with any degree of NAFLD. NAS of > or =5 correlated with a diagnosis of NASH, and biopsies with scores of less than 3 were diagnosed as "not NASH."
Nonalcoholic fatty liver disease (NAFLD) is one of the most frequent causes of abnormal liver dysfunction, and its prevalence has markedly increased; however, the mechanisms involved in the pathogenesis of NAFLD have not been thoroughly investigated in humans. In this study, we evaluated the expression of fatty acid metabolism-related genes in NAFLD. Real-time RT-PCR was performed using liver biopsy samples from 12 NAFLD patients. The target genes studied were: acetyl-CoA carboxylase (ACC) 1, ACC2, and fatty acid synthase (FAS) for the evaluation of de novo fatty acid synthesis; carnitine palmitoyltransferase 1a (CPT1a), long-chain acyl-CoA dehydrogenase (LCAD), and long-chain L-3-hydroxyacyl-coenzyme A dehydrogenase alpha (HADHalpha) for beta-oxidation in the mitochondria; peroxisome proliferator-activated receptor- (PPAR-) alpha and cytochrome P450 2E1 (CYP2E1) for oxidation in peroxisomes and microsomes (endoplasmic reticulum) respectively; and diacylglycerol O-acyltransferase 1 (DGAT1), PPAR-gamma, and hormone sensitive lipase (HSL) for triglyceride synthesis and catalysis. In NAFLD, expression of ACC1 and ACC2, but not FAS was increased, indicating that de novo fatty acid synthesis is enhanced in NAFLD. In contrast, expression of CTP1a, a rate-limiting enzyme, was remarkably decreased, indicating that beta-oxidation in the mitochondria was decreased, although the expression of LCAD and HADHalpha was increased. Expression of PPAR-alpha was increased, whereas that of CYP2E1 was reduced. The expression of DGAT1, PPAR-gamma, and HSL was enhanced. These data suggest that in NAFLD, increased de novo synthesis and decreased beta-oxidation in the mitochondria lead to accumulation of fatty acids in hepatocytes, although the extent of oxidation in peroxisomes and microsomes remains unclear.
Oxidative DNA damage is considered to play an important role in pathophysiological processes, ageing and cancer. So far major interest has been on measuring 8-hydroxy-2'-deoxyguanosine (8-OHdG), the preferred methods relying on HPLC or GC-mass spectrometry. The high biological relevance of 8-OHdG is due to its ability to induce G-->T transversions, which are among the most frequent somatic mutations found in human cancers. Effects of workplace exposures on the level of white blood cell 8-OHdG or urinary 8-OHdG have been reported with controversial results. Exposures examined include asbestos, azo-dyes, benzene, fine particulate matter (PM(2.5)), glassworks, polycyclic aromatic hydrocarbons (PAHs), rubber manufacturing, silica, metals, styrene, toluene and xylenes. The available data indicate that there is still a lack of well established dose-response relations between occupational or environmental exposures and the induction of 8-OHdG. Smoking has been most consistently identified as a confounder for 8-OHdG, but various occupational studies did not reveal higher levels of 8-OHdG in smokers. Despite the conflicting results, the reported studies show promise for 8-OHdG as a biomarker of oxidative stress associated with chemical exposure. However, there are critical aspects related to the analytical challenge, artifactual production of 8-OHdG, inter- and intra-individual variation, confounding factors and inter-laboratory differences, implying that further work is needed to reach a consensus on the background level of 8-OHdG.
C-Reactive Protein (CRP), a nonspecific marker of inflammation that is moderately elevated in obesity, metabolic syndrome (MS), and type 2 diabetes, has been proposed as a surrogate marker of nonalcoholic steatohepatitis (NASH). Its clinical usefulness in the diagnosis of NASH was evaluated in severely obese patients without or with MS, diabetes, and NASH and the potential roles of the liver and of the adipose tissue in CRP production were characterized.
Severely obese patients without NASH (without MS [N = 13], with MS [N = 11], or with MS and diabetes [N = 7]) and with NASH (without [N = 8] or with [N = 7] MS) were studied. For each patient, liver and adipose tissue biopsies were collected during a bariatric surgery and were used to determine the CRP gene expression by real-time PCR. The role of interleukin-6 (IL6) and lipopolysaccharide in CRP expression was also evaluated in subcutaneous adipose tissue obtained during cosmetic abdominoplasty.
Plasma CRP levels were elevated in severely obese patients independently from the presence or absence of MS, diabetes, or NASH. CRP gene expression was not only increased in livers but also in adipose tissues of obese patients compared with controls subjects. In human adipose tissue, CRP mRNA levels were positively correlated with those of IL-6 and the CRP expression was enhanced in vitro by IL-6 and lipopolysaccharide.
Plasma CRP levels are not predictive of the diagnosis of NASH in severely obese patients. The liver but also the adipose tissue can produce CRP, a process which could be dependent on IL6. Therefore, both tissues might contribute to the elevated plasma CRP levels found in obesity. In addition, the large amount of body fat may well produce an important part of the circulating CRP, further limiting its clinical usefulness in the evaluation of NASH in severely obese patients.
Insulin resistance (IR) is commonly associated with non-alcoholic steatohepatitis (NASH). Peroxisome proliferator-activated receptor-alpha (PPAR-alpha) may also play a role in the pathogenesis of NASH. A pivotal role in NASH pathogenesis depends on the hypothesis of increased oxidative stress. The aim of our study was to evaluate the effects of supplemental oral vitamin E, a potent antioxidant, on liver functions, PPAR-alpha expression and IR in patients with NASH.
Nine patients with biopsy-proven NASH were given oral vitamin E 800 mg daily for 24 weeks. Liver functions, lipid parameters, IR index with homeostatic metabolic assessment and liver histology and PPAR-alpha staining index in biopsy specimens were detected before and after the treatment.
Seven patients (78%) had IR initially. After 6 months of therapy in nine patients, fasting insulin improved (P = 0.01), but serum cholesterol, triglyceride, fasting blood glucose levels and body mass index remained unchanged. Aspartate aminotransferase and alanine aminotransferase levels decreased (P = 0.01 and P = 0.01, respectively). IR index with homeostatic metabolic assessment resistance improved (P = 0.01), but PPAR-alpha staining index did not change (P = 0.37). Although the histological grade of steatosis decreased (P = 0.01), necroinflammation and fibrosis remained unchanged. In seven patients with IR, however, necroinflammation and PPAR-alpha staining index were improved (P = 0.04 and P = 0.02).
Vitamin E treatment, in addition to its previously shown beneficial effect by suppressing oxidative stress, may also achieve improvement by reducing IR and PPAR-alpha expression in NASH.
Mitochondrial dysfunction is involved in the three stages of the transition from lack of exercise and excessive food intake to insulin resistance, diabetes and non-alcoholic steatohepatitis (NASH). In muscle, lack of exercise, a fat-rich diet, a polymorphism in peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1), and possibly age-related mitochondrial DNA (mtDNA) mutations may variously combine their effects to decrease PGC-1 expression, mitochondrial biogenesis and fat oxidation. Together with excessive food intake, insufficient fat oxidation causes fat accumulation and cellular stress in myocytes. The activation of Jun N-terminal kinase and protein kinase C-theta triggers the serine phosphorylation and inactivation of the insulin receptor substrate, and hampers the insulin-mediated translocation of glucose transporter-4 to the plasma membrane. Initially, the trend for increased blood glucose increases insulin secretion by pancreatic beta-cells. High plasma insulin levels compensate for insulin resistance in muscle and maintain normal blood glucose levels. Eventually, however, increased uncoupling protein-2 expression and possibly acquired mtDNA mutations in pancreatic beta-cells can blunt glucose-mediated adenosine triphosphate (ATP) formation and insulin secretion, to cause diabetes in some patients. High plasma glucose and/or insulin levels induce hepatic lipogenesis and cause hepatic steatosis. In fat-engorged hepatocytes, several vicious cycles involving tumor necrosis factor-alpha, reactive oxygen species (ROS), peroxynitrite, and lipid peroxidation products alter respiratory chain polypeptides and mtDNA, thus partially blocking the flow of electrons in the respiratory chain. The overreduction of upstream respiratory chain complexes increases mitochondrial ROS and peroxynitrite formation. Oxidative stress increases the release of lipid peroxidation products and cytokines, which together trigger the liver lesions of NASH.
Nonalcoholic fatty liver disease (NAFLD) is one of the most frequent causes of abnormal liver dysfunction, and its prevalence has markedly increased. We previously evaluated the expression of fatty acid metabolism-related genes in NAFLD and reported changes in expression that could contribute to increased fatty acid synthesis. In the present study, we evaluated the expression of additional fatty acid metabolism-related genes in larger groups of NAFLD (n=26) and normal liver (n=10) samples. The target genes for real-time PCR analysis were as follows: acetyl-CoA carboxylase (ACC) 1, ACC2, fatty acid synthase (FAS), sterol regulatory element-binding protein 1c (SREBP-1c), and adipose differentiation-related protein (ADRP) for evaluation of de novo synthesis and uptake of fatty acids; carnitine palmitoyltransferase 1a; (CPT1a), long-chain acyl-CoA dehydrogenase (LCAD), long-chain L-3-hydroxyacylcoenzyme A dehydrogenase alpha (HADHalpha), uncoupling protein 2 (UCP2), straight-chain acyl-CoA oxidase (ACOX), branched-chain acyl-CoA oxidase (BOX), cytochrome P450 2E1 (CYP2E1), CYP4A11, and peroxisome proliferator-activated receptor (PPAR)alpha for oxidation in the mitochondria, peroxisomes and microsomes; superoxide dismutase (SOD), catalase, and glutathione synthetase (GSS) for antioxidant pathways; and diacylglycerol O-acyltransferase 1 (DGAT1), PPARgamma, and hormone-sensitive lipase (HSL) for triglyceride synthesis and catalysis. In NAFLD, although fatty acids accumulated in hepatocytes, their de novo synthesis and uptake were up-regulated in association with increased expression of ACC1, FAS, SREBP-1c, and ADRP. Fatty acid oxidation-related genes, LCAD, HADHalpha, UCP2, ACOX, BOX, CYP2E1, and CYP4A11, were all overexpressed, indicating that oxidation was enhanced in NAFLD, whereas the expression of CTP1a and PPARalpha was decreased. Furthermore, SOD and catalase were also overexpressed, indicating that antioxidant pathways are activated to neutralize reactive oxygen species (ROS), which are overproduced during oxidative processes. The expression of DGAT1 was up-regulated without increased PPARgamma expression, whereas the expression of HSL was decreased. Our data indicated the following regarding NAFLD: i) increased de novo synthesis and uptake of fatty acids lead to further fatty acid accumulation in hepatocytes; ii) mitochondrial fatty acid oxidation is decreased or fully activated; iii) in order to complement the function of mitochondria (beta-oxidation), peroxisomal (beta-oxidation) and microsomal (omega-oxidation) oxidation is up-regulated to decrease fatty acid accumulation; iv) antioxidant pathways including SOD and catalase are enhanced to neutralize ROS overproduced during mitochondrial, peroxisomal, and microsomal oxidation; and v) lipid droplet formation is enhanced due to increased DGAT expression and decreased HSL expression. Further studies will be needed to clarify how fatty acid synthesis is increased by SREBP-1c, which is under the control of insulin and AMP-activated protein kinase.