Article

Hydrogen inhalation ameliorates lipopolysaccharide-induced acute lung injury in mice

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Abstract

Acute lung injury (ALI) is a serious illness, the incidence and mortality of which are very high. Free radicals, such as hydroxyl radicals (OH) and peroxynitrite (ONOO(-)), are considered to be the final causative molecules in the pathogenesis of ALI. Hydrogen, a new antioxidant, can selectively reduce OH and ONOO(-). In the present study, we investigated the hypothesis that hydrogen inhalation could ameliorate ALI induced by intra-tracheal lipopolysaccharide (LPS, 5mg/kg body weight). Mice were randomized into three groups: sham group (physiological saline+2% hydrogen mixed gas), control group (LPS+normal air) and experiment group (LPS+2% hydrogen mixed gas). Bronchoalveolar lavage fluid (BALF) was performed to determine the total protein concentrations and pro-inflammatory cytokines. Lung tissues were assayed for oxidative stress variables, wet/dry (W/D) ratio, histological, immunohistochemistry and Western blotting examinations. Our experiments exhibited that hydrogen improved the survival rate of mice and induced a decrease in lung W/D ratio. In addition, hydrogen decreased malonaldehyde and nitrotyrosine content, inhibited myeloperoxidase and maintained superoxide dismutase activity in lung tissues and associated with a decrease in the expression of TNF-α, IL-1β, IL-6 and total protein concentrations in the BALF. Hydrogen further attenuated histopathological alterations and mitigated lung cell apoptosis. Importantly, hydrogen inhibited the activation of P-JNK, and also reversed changes in Bax, Bcl-xl and caspase-3. In conclusion, our data demonstrated that hydrogen inhalation ameliorated LPS-induced ALI and it may be exerting its protective role by preventing the activation of ROS-JNK-caspase-3 pathway.

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... Lipopolysaccharide (LPS) is a component of the cell wall of Gram-negative bacteria and the most important pathogenic factor in sepsis. It has been demonstrated that molecular hydrogen extenuates LPS-induced ALI in rats by reducing the release of inflammatory factors, inhibiting the aggregation of inflammatory cells, reducing oxidative stress and apoptosis [8,11,13]. Moreover, molecular hydrogen alleviates pulmonary edema caused by LPS through upregulating the expression of pulmonary aquaporin (AQP) [14]. ...
... Molecular hydrogen is a natural antioxidant that antagonizes oxidative stress in several ways ( Fig. 3): (1) by neutralizing ·OH [54], (2) reducing ·ONOO-and its gene expression directly [55], and inhibiting the production of nitro-tyrosine indirectly, which is an indicator of ·ONOO-generation [13], (3) by inducing antioxidant gene expression and increasing antioxidant enzyme activity, including superoxide dismutase (SOD), HO-1, catalase (CAT), and myeloperoxidase (MPO) [56], (4) by reducing the levels of oxidative stress indicators, such as 8-iso-prostaglandin F2α [57], and the lipid peroxidation marker malondialdehyde (MDA) [13], (5) by reducing ·NO production through inhibiting inducible nitric oxide synthase (iNOS) [29,58] and endothelial nitric oxide synthase (eNOS) [59], and (6) by inhibiting NADPH oxidase activity, which is the main source of free radicals in sepsis [59][60][61]. ...
... Molecular hydrogen is a natural antioxidant that antagonizes oxidative stress in several ways ( Fig. 3): (1) by neutralizing ·OH [54], (2) reducing ·ONOO-and its gene expression directly [55], and inhibiting the production of nitro-tyrosine indirectly, which is an indicator of ·ONOO-generation [13], (3) by inducing antioxidant gene expression and increasing antioxidant enzyme activity, including superoxide dismutase (SOD), HO-1, catalase (CAT), and myeloperoxidase (MPO) [56], (4) by reducing the levels of oxidative stress indicators, such as 8-iso-prostaglandin F2α [57], and the lipid peroxidation marker malondialdehyde (MDA) [13], (5) by reducing ·NO production through inhibiting inducible nitric oxide synthase (iNOS) [29,58] and endothelial nitric oxide synthase (eNOS) [59], and (6) by inhibiting NADPH oxidase activity, which is the main source of free radicals in sepsis [59][60][61]. ...
Article
Sepsis is a syndrome comprised of a series of life-threatening organ dysfunctions caused by a maladjusted body response to infection with no effective treatment. Molecular hydrogen is a new type of antioxidant with strong free radical scavenging ability, which has been demonstrated to be effective for treating various diseases, such as infection, trauma, poisoning, organ ischemia-reperfusion, metabolic diseases, and tumors. Molecular hydrogen exerts multiple biological effects involving anti-inflammation, anti-oxidation, anti-apoptosis, anti-shock, and autophagy regulation, which may attenuate the organ and barrier damage caused by sepsis. However, the underlying molecular mechanisms remain elusive, but are likely related to the signaling pathways involved. This review focuses on the research progress and potential mechanisms of molecular hydrogen against sepsis to provide a theoretical basis for clinical treatment.
... The clinical manifestations include hypoxemia, increased respiratory rate, and bi-pneumal diffuse infiltration in X-ray. The mortality rate of ALI was as high as 39% [1][2][3][4][5][6], and certain studies have shown that neutrophils played important roles in the pathogenesis of ALI [7][8][9], the activation of which would release oxygen free radicals, proteases, leukotriene, and other proinflamma-tory molecules, thus initiating the inflammatory cascade, and eventually leading to ALI. An in vitro study has shown that alkalizing blood could weaken neutrophil's adhesion and accelerate their apoptosis [10][11][12]; however, blood alkalization treatment for ALI has not been reported in vivo thus far. ...
... After the experiment, the left lung of a mouse was collected to prepare 10% lung tissue homogenate, which was then centrifuged and the supernatant was evaluated for MDA and MPO content [7][8][9][10]. ...
... compared with group normal saline, group NaHCO 3 had significantly elevated plasma [Na + ] and decreased [K + ] (P<0.05), while group THAM had significantly reduced plasma [Na + ] and elevated plasma lactic acid concentration (P<0.05). However, pCO 2 , pO 2 , blood glucose wet weight was measured, tissues were incubated in 90°C oven to constant weight, the dry weight was measured, and then, the wet/dry weight ratio was calculated [7][8][9][10]. ...
Article
The aim of this study was to investigate the effects of basic drugs that alkalizes blood, on prognosis of acute lung injury in mice. Mice were randomized into three groups: Group normal saline, Group THAM, injected with 3.64% tri-(hydroxymethyl) methylamine (THAM), and Group NaHCO3, injected with 5% NaHCO3 (n=26, each group). The acute lung injury model was established by intraperitoneal injection of lipopolysaccharide (LPS; 50 mg/kg), followed by infusion of varying concentrations of the above solution into tail vein at the rate of 0.5 ml/h (controlled by micro pump) for over 2 h. Thirty minutes later, 6 mice from each group were randomly selected for blood gas analysis; then, the mice were killed and their lung tissues were sampled for detection of relative indicators, and the remaining mice were observed for signs of mortality for 72 h. Arterial pH, bicarbonate (HCO3-), and BE and mortality of group THAM and NaHCO3 increased significantly compared to the corresponding parameters of the group normal saline (P<0.05); compared to the group normal saline, group NaHCO3 had increased blood [Na+] and decreased [K+] and [Ca2+] (P<0.05). Blood [Na+] of group THAM decreased while the lactic acid concentration increased (P<0.05) compared to the corresponding values of the group normal saline. Malondialdehyde (MDA) and myeloperoxidase (MPO) activity and wet-to-dry lung weight ratio (W/D) of group THAM and NaHCO3 increased significantly relative to group normal saline (P<0.05). Compared with the biopsy results of (A), pathological biopsy of (B) and (C) clearly revealed alveolar wall thickening, edema of alveolar epithelial cells, and infiltration of large neutrophils. Alkalizing blood could neither inhibit inflammatory reactions in LPS mouse model nor reduce the mortality rate of mice with acute lung injury, while excessive alkalization of blood could increase mice mortality.
... Despite the high morbidity and mortality rates of this illness, mechanisms underlying the development of ARDS/ALI remain incompletely understood. [8][9][10][11] Respiratory distress syndrome (RDS) in neonates occurs due to surfactant deficiency and immaturity of the lung parenchyma and vasculature. The course of RDS is characterized by parenchymal lung injury, which leads to impaired gas exchange, neutrophil accumulation in the lung, the expression of pro-inflammatory mediators, increased vascular permeability and damage to the lung epithelium and endothelium. ...
... One of the contributing factors in the development of ALI in premature babies is exposure to bacterial infection, including endotoxin from gram-negative bacteria. [8][9][10][12][13][14] Exposure to supplemental oxygen, which is often used in the treatment of RDS, can also contribute to lung injury. ...
... Decreased anti-oxidant capacity of pulmonary vascular tissue along with increased production of ROS contributes to the injury seen in LPS-induced ALI/BPD. 7,9,[23][24][25][26][27] ROS generated by LPS exposure in endothelial cells is regarded as a key to the modulation of the pulmonary vascular endothelial damage, which leads to higher oxidative stress. Increased ROS may cause cell injury, activate the inflammatory response, promote cytotoxicity and activate signaling pathways that lead to pro-apoptotic signaling. ...
Article
Full-text available
Reactive oxygen species (ROS) have been implicated in the pathogenesis of many acute and chronic pulmonary disorders such as acute lung injury (ALI) in adults and bronchopulmonary dysplasia (BPD) in premature infants. Bacterial infection and oxygen toxicity, which result in pulmonary vascular endothelial injury, contribute to impaired vascular growth and alveolar simplification seen in the lungs of premature infants with BPD. Hyperoxia induces ALI, reduces cell proliferation, causes DNA damage and promotes cell death by causing mitochondrial dysfunction. The objective of this study was to use an optical imaging technique to evaluate the variations in fluorescence intensities of the auto-fluorescent mitochondrial metabolic coenzymes, NADH and FAD in four different groups of rats. The ratio of these fluorescence signals (NADH/FAD), referred to as NADH redox ratio (NADH RR) has been used as an indicator of tissue metabolism in injuries. Here, we investigated whether the changes in metabolic state can be used as a marker of oxidative stress caused by hyperoxia and bacterial lipopolysaccharide (LPS) exposure in neonatal rat lungs. We examined the tissue redox states of lungs from four groups of rat pups: normoxic (21% O2) pups, hyperoxic (90% O2) pups, pups treated with LPS (normoxic + LPS), and pups treated with LPS and hyperoxia (hyperoxic + LPS). Our results show that hyperoxia oxidized the respiratory chain as reflected by a ~31% decrease in lung tissue NADH RR as compared to that for normoxic lungs. LPS treatment alone or with hyperoxia had no significant effect on lung tissue NADH RR as compared to that for normoxic or hyperoxic lungs, respectively. Thus, NADH RR serves as a quantitative marker of oxidative stress level in lung injury caused by two clinically important conditions: hyperoxia and LPS exposure.
... Oxidative stress, mitochondrial dysfunction, and inflammation are key pathways in the pathogenesis of ARDS or ALI, with the pulmonary capillary endothelium a primary and early target (17)(18)(19)(20)(21)(22)(23). A large body of evidence supports that the pulmonary capillary endothelium, which has a large surface area and is in close contact with blood-borne compounds, is a primary and early site of injury (17)(18)(19)(20), which can be targeted in vivo with single-photon emission computed tomography (SPECT) (24)(25)(26)(27)(28)(29). ...
... Oxidative stress, mitochondrial dysfunction, and inflammation are key pathways in the pathogenesis of ARDS or ALI, with the pulmonary capillary endothelium a primary and early target (17)(18)(19)(20)(21)(22)(23). A large body of evidence supports that the pulmonary capillary endothelium, which has a large surface area and is in close contact with blood-borne compounds, is a primary and early site of injury (17)(18)(19)(20), which can be targeted in vivo with single-photon emission computed tomography (SPECT) (24)(25)(26)(27)(28)(29). Previously, we demonstrated the utility of in vivo 99m Tc-hexamethylpropyleneamine oxime (HMPAO) and 99m Tc-duramycin imaging to detect oxidative stress and endothelial cell death in rats exposed to hyperoxia, respectively (24)(25)(26)(27)(28)(29). ...
Article
99m Tc-hexamethylpropyleneamine oxime (HMPAO) and 99m Tc-duramycin in vivo imaging detects pulmonary oxidative stress and cell death, respectively, in rats exposed to >95% O 2 (hyperoxia) as a model of Acute Respiratory Distress Syndrome (ARDS). Pre-exposure to hyperoxia for 48 h followed by 24 h in room air (H-T) is protective against hyperoxia-induced lung injury. This study's objective was to determine the ability of 99m Tc-HMPAO and 99m Tc-duramycin to track this protection and to elucidate underlying mechanisms. Rats were exposed to normoxia, hyperoxia for 60 h, H-T, or H-T followed by 60 h of hyperoxia (H-T+60). Imaging was performed 20 minutes post intravenous injection of either 99m Tc-HMPAO or 99m Tc-duramycin. 99m Tc-HMPAO and 99m Tc-duramycin lung uptake was 200% and 167% greater (p <0.01) in hyperoxia compared to normoxia rats, respectively. On the other hand, uptake of 99m Tc-HMPAO in H-T+60 was 24% greater (p <0.01) than in H-T rats, but 99m Tc-duramycin uptake was not significantly different (p=0.09). Lung wet-to-dry weight ratio, pleural effusion, endothelial filtration coefficient, and histological indices all showed evidence of protection and paralleled imaging results. Additional results indicate higher mitochondrial complex IV activity in H-T versus normoxia rats, suggesting that mitochondria of H-T lungs may be more tolerant of oxidative stress. A pattern of increasing lung uptake of 99m Tc-HMPAO and 99m Tc-duramycin correlates with advancing oxidative stress and cell death and worsening injury, whereas stable or decreasing 99m Tc-HMPAO and stable 99m Tc-duramycin reflects hyperoxia tolerance, suggesting the potential utility of molecular imaging for identifying at-risk hosts that are more or less susceptible to progressing to ARDS.
... 38 Qiu et al. showed that lipopolysaccharide (LPS) resulted in an upregulation of Bax protein and hydrogen inhalation inhibited overactivity of Bax protein. 39 These influences of hydrogen on apoptosis-related genes can reduce the secretion of proteases, protect the integrity of cells from damage, and inhibit the initiation of apoptosis programs. 13 Hydrogen protects mitochondrial structure and function in I/R injury Mitochondria is the major site of generation of intracellular ROS as well as the target of ROS. ...
... 37 Qiu et al. found that hydrogen can inhibit the production of ROS in lung tissue caused by endotoxins and inhibit the activation of p-JNK. 39 Bai et al. proved that saturated hydrogen water could reduce the occurrence of the inflammatory cascade and organ dysfunction by inhibiting the ERK1/2 pathway. 52 Intraperitoneal injection of HRS inhibited LPS-induced activation of p38 MAPK and JNK, which is associated with LPS-induced downregulation of aquaporin-1 (AQP-1) andaquaporin-5 (AQP-5). ...
Article
Lung grafts may experience multiple injuries during lung transplantation, such as warm ischaemia, cold ischaemia, and reperfusion injury. These injuries all contribute to primary graft dysfunction, which is a major cause of morbidity and mortality after lung transplantation. As a potential selective antioxidant, hydrogen molecule (H2) protects against post-transplant complications in animal models of multiple organ transplantation. Herein, the authors review the current literature regarding the effects of H2 on lung injury from lung transplantation. The reviewed studies showed that H2 improved the outcomes of lung transplantation by decreasing oxidative stress and inflammation at the donor and recipient phases. H2 is primarily administered via inhalation, drinking hydrogen-rich water, hydrogen-rich saline injection, or a hydrogen-rich water bath. H2 favorably modulates signal transduction and gene expression, resulting in the suppression of pro-inflammatory cytokines and excess reactive oxygen species production. Although H2 appears to be a physiological regulatory molecule with antioxidant, anti-inflammatory and anti-apoptotic properties, its exact mechanisms of action remain elusive. Taken together, accumulating experimental evidence indicates that H2 can significantly alleviate transplantation-related lung injury, mainly via inhibition of inflammatory cytokine secretion and reduction in oxidative stress through several underlying mechanisms. Further animal experiments and preliminary human clinical trials will lay the foundation for the use of H2 as a treatment in the clinic.
... These studies have confirmed the data reported in 2007 [28] that H 2 acts directly on • OH and ONOO _ to decrease oxidative stress. Furthermore, recent studies have added other ways in which H 2 may reduce oxidative stress such as (i) regulating antioxidant gene expression and rising antioxidant enzyme activity such as SOD, CAT, myeloperoxidase (MPO) and heme oxygenase-1(HO-1) [32]; (ii) decreasing oxidative stress status such as 8-iso-prostaglandin-F2α (8-iso-PGF2α) [33], thiobarbituric acid reactive substances (TBARS) [34] and malondialdehyde (MDA) [35]; (iv) reducing nitric oxide (NO) production [33,36,37]; and (v) inhibiting nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity [37][38][39]. The potential effects of H 2 are summarized in Fig. (1). ...
... Recent studies indicate that H 2 down-modulates the activation of mitogenactivated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling pathways [40,41], as well as pro-inflammatory cytokines such as TNF-α, interleukin IL-1β, IL-6 [15,16,19,24], interferon-γ (IFN-γ) [42] and high mobility group box 1 protein (HMGB1) [43]. Furthermore, H 2 has been documented to act (i) up-modulating the anti-inflammatory cytokine IL-10 [16,19,29]; (ii) reducing chemokines macrophage inflammatory protein 1 and 2 (MIP1 and MIP2) [44]; and (iii) decreasing neutrophils [35], lymphocytes [45] and macrophages [44,[46][47][48]. These data are summarized in Fig. (2). ...
Article
Physical exercise-induced oxidative stress and inflammation may be beneficial when exercise is a regular activity, but it is rather harmful when exercise is exhaustive and performed by unaccustomed organisms. Molecular hydrogen (H2 ) has recently appeared as a potent antioxidant and anti-inflammatory molecule in numerous pathological conditions. However, its role is relatively unknown under physiological conditions such as physical exercise. Therefore, this review summarizes the current knowledge of the H2 reducing oxidative stress and inflammation in physical exercise, reporting data from both animal and human studies.
... Data are shown as mean + SEM with squares representing values from individual mice. c-g Total cell numbers in the BALF (c), representative BALF cell cytocentrifuge preparations stained with diff-quik (neutrophils are indicated by yellow arrows) (d), BALF neutrophil numbers (e), representative flow cytometry profile of neutrophils (Gr-1 + CD11b + ) in dissociated lung tissues (f) and numbers of neutrophils (Gr-1 + CD11b + ) in the lungs (g) of Tslpr +/+ mice (n = 10-18) and Tslpr −/− mice (n = [19][20]. h-m Hematoxylin and eosin (HE) histopathology showing that a Tslpr +/+ mouse treated with saline (h, k) had no changes in its bronchioles or bronchiolar lumens (Br), vessels (V), or alveolar spaces (a). ...
... However, we did not observe a significant reduction in lymphocyte numbers in the BALF from Tslpr −/− mice when compared with Tslpr +/+ mice ( Fig. S7) suggesting that the effects of TSLP on the lymphoid compartment do not contribute to protection from bleomycin-induced airway inflammation. There is extensive literature supporting a protective role for Bcl-2 and Bcl-xL in models of acute lung injury, including bleomycininduced damage and inflammation [18][19][20][21][22] . In concordance with this evidence, Tslpr −/− mice exhibited significantly lower mRNA expression levels for Bcl-xL (Fig. 3h) while a slight, but not significant, increase in Bcl-2 mRNA expression levels ( Fig. 3i) was observed when compared with Tslpr +/+ mice treated with bleomycin. ...
Article
Full-text available
Thymic stromal lymphopoietin (TSLP), an epithelial cell-derived cytokine, exhibits both pro-inflammatory and pro-homeostatic properties depending on the context and tissues in which it is expressed. It remains unknown whether TSLP has a similar dual role in the airways, where TSLP is known to promote allergic inflammation. Here we show that TSLP receptor (TSLPR)-deficient mice (Tslpr−/−) and mice treated with anti-TSLP antibodies exhibited increased airway inflammation and morbidity rates after bleomycin-induced tissue damage. We found that signaling through TSLPR on non-hematopoietic cells was sufficient for TSLP’s protective function. Consistent with this finding, we showed that TSLP reduces caspase-1 and caspase-3 activity levels in primary human bronchial epithelial cells treated with bleomycin via Bcl-xL up-regulation. These observations were recapitulated in vivo by observing that Tslpr−/− mice showed reduced Bcl-xL expression that paralleled increased lung caspase-1 and caspase-3 activity levels and IL-1β concentrations in the bronchial-alveolar lavage fluid. Our studies reveal a novel contribution for TSLP in preventing damage-induced airway inflammation.
... Hydrogen gas (H 2 ), a medical gas that has anti-oxidant, anti-apoptotic, and anti-inflammatory properties (Ohsawa et al., 2007;Yu et al., 2011;Zhai et al., 2013;Zhai et al., 2014;Li et al., 2016;Zhai et al., 2017), has been reported to improve sepsis-induced organ dysfunction, such as lung (Qiu et al., 2011;Liang et al., 2012;Xie et al., 2012;Hattori et al., 2015;Liu et al., 2015;Dong et al., 2017;Dong et al., 2018), liver Iketani et al., 2017), and bowel (Sakata et al., 2017). Recent studies have also indicated that H 2 has strong cardiovascular activities (Zhang et al., 2018). ...
... Therefore, blocking TLR4-related innate immune signaling can improve the previously discussed stimuli-mediated cardiac dysfunction. H 2 has been shown to improve LPS-induced lung (Qiu et al., 2011;Liang et al., 2012;Xie et al., 2012;Hattori et al., 2015;Liu et al., 2015;Dong et al., 2017), liver Iketani et al., 2017), and bowel (Sakata et al., 2017) injuries. Herein, we showed that H 2 inhibited LPS-mediated cardiac dysfunction, which can be induced by LPS and LPS-induced many kinds of cytokines, such as TNFα, IL-1β, and IL-18 (Thaik et al., 1995;Bryant et al., 1998;Li et al., 2002;Carlson et al., 2005;Suzuki et al., 2007;Liu et al., 2008;Wu et al., 2009;Chowdhury et al., 2013;Drosatos et al., 2013;Okuhara et al., 2017). ...
Article
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Background and Purpose: Septic cardiomyopathy, which is one of the features of multi-organ dysfunction in sepsis, is characterized by ventricular dilatation, reduced ventricular contractility, and reduction in ejection fraction and, if severe, can lead to death. To date, there is no specific therapy that exists, and its treatment represents a large unmet clinical need. Herein, we investigated the effects and underlying anti-inflammatory mechanisms of hydrogen gas in the setting of lipopolysaccharide (LPS)-induced cardiomyocytes injury. Experimental Approach: Hydrogen gas was intraperitoneally injected to mice in LPS plus hydrogen group and hydrogen group for 4 days. On fourth, LPS was given by intraperitoneal injection to mice in LPS group and to mice in LPS plus hydrogen group. In addition, H9c2 cardiomyocytes were treated with hydrogen-rich medium for 30 min before LPS. The transthoracic echocardiography was performed at 6 h post‐LPS to assess left ventricular end-systolic diameter (LVESD), left ventricular end-diastolic diameter (LVEDD), left ventricular ejection fraction (EF%), fractional shortening (FS%), left ventricular mass average weight (LV mass AW), and LV mass AW (Corrected). The histological and morphological analyses of left ventricular were performed by hematoxylin and eosin (H&E) staining and Masson’s trichrome staining. The mRNA levels of ANP and BNP were examined by PCR in vitro. The expression of cytokines were assayed by Enzyme Linked Immunosorbent Assay (ELISA) and PCR. Moreover, Western blotting was performed to examine the expression of TLR4, the activation of ERK1/2, p38, JNK, and the expression of NF-κB in nucleus after 6 h of LPS challenge in vivo and in vitro. Key Results: LPS induced cardiac dysfunction; hydrogen therapy improved cardiac function after LPS challenge. Furthermore, pretreatment with hydrogen resulted in cardioprotection during septic cardiomyopathy via inhibiting the expression of pro-inflammatory cytokines TNFα, IL-1β, and IL-18; suppressing the phosphorylation of ERK1/2, p38, and JNK; and reducing the nuclear translocation of NF-κB and the expression of TLR4 by LPS. Conclusion and Implications: Hydrogen therapy prevents LPS-induced cardiac dysfunction in part via downregulation of TLR4-mediated pro-inflammatory cytokines expression.
... Furthermore, Yasuhiro et al. illustrated that hydrogen could prevent radioactive lung injury by inhibiting oxidative stress at the cellular level [21]. In addition, two studies found that hydrogen gas inhalation could improve lipopolysaccharide-induced lung injury by prolonging animal survival times, reducing oxidative stress damage, and inhibiting cell apoptosis [22,23]. The present study reveals that hydrogen gas inhalation significantly improves hypoxemia and lung histopathologic changes, alleviates pulmonary edema and lung endothelial permeability, prevents the release of TNF-α, IL-1β, Fig. 6. ...
... Second, we acknowledge that the animal model of seawater instillation-induced acute lung injury, like most animal models, does not completely recapitulate all aspects of seawater instillation-induced acute lung injury in a clinical setting. Lastly, although hydrogen has no risk of explosion at concentrations less than 4 and 2 % hydrogen was used in most previous studies [22], the best concentration of hydrogen gas inhalation needs to be explored in the future studies. ...
Article
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Seawater instillation-induced acute lung injury involves oxidative stress and apoptosis. Although hydrogen gas inhalation is reportedly protective in multiple types of lung injury, the effect of hydrogen gas inhalation on seawater instillation-induced acute lung injury remains unknown. This study investigated the effect of hydrogen gas on seawater instillation-induced acute lung injury and explored the mechanisms involved. Rabbits were randomly assigned to control, hydrogen (2 % hydrogen gas inhalation), seawater (3 mL/kg seawater instillation), and seawater + hydrogen (3 mL/kg seawater instillation + 2 % hydrogen gas inhalation) groups. Arterial partial oxygen pressure and lung wet/dry weight ratio were detected. Protein content in bronchoalveolar lavage fluid (BALF) and serum as well as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 levels were determined. Hematoxylin-eosin staining was used to monitor changes in lung specimens, and malondialdehyde (MDA) content and myeloperoxidase (MPO) activity were assayed. In addition, NF-E2-related factor (Nrf) 2 and heme oxygenase (HO)-1 mRNA and protein expression were measured, and apoptosis was assessed by measuring caspase-3 expression and using terminal deoxy-nucleotidyl transferase dUTP nick end-labeling (TUNEL) staining. Hydrogen gas inhalation markedly improved lung endothelial permeability and decreased both MDA content and MPO activity in lung tissue; these changes were associated with decreases in TNF-α, IL-1β, and IL-6 in BALF. Hydrogen gas also alleviated histopathological changes and cell apoptosis. Moreover, Nrf2 and HO-1 expressions were significantly activated and caspase-3 expression was inhibited. These results demonstrate that hydrogen gas inhalation attenuates seawater instillation-induced acute lung injury in rabbits and that the protective effects observed may be related to the activation of the Nrf2 pathway.
... On the other hand, a collation of hydrogen reports indicate that a similar degree of effects can be observed with different modalities of administration. For example, the marked effect of hydrogen on a mouse model of LPS-induced acute lung injury has been reported by four different groups with three different modalities: hydrogen gas[13,14], hydrogen water[15], and hydrogen-rich saline[14,16]. Similarly, the dramatic effect of hydrogen on animal models of acute myocardial infarction has been reported by eight different groups with two different modalities: hydrogen gas[17][18][19][20]and hydrogen-rich saline[21][22][23][24]. ...
... Hearing loss[143][144][145][146][147][148]Cisplatin-induced ototoxicity[149,150]Ouabain-induced ototoxicity[151]Oral Cavity Periodontitis[32]Periodontal oxidative damage[152]Lung Lung I/R injury[153,154]Oxygen-induced lung injury[82,155,156]Ventilation-induced lung injury[53,157]LPS-induced acute lung injury[13,14,16,158]Intestinal I/R-induced lung injury[159]Burn-induced lung injury[160]Paraquat-induced lung injury[161,162]igarette smoking lung injury[163]Smoke inhalation lung injury[74]Pulmonary hypertension[78,164]Heart Myocardial infarction and I/R injury[17][18][19][20][21][22][23][24]84]Diabetic cardiomyopathy[40]Sleep apnea-induced left ventricular remodeling[165,166]Ventricular hypertrophy[167]Stomach Stress-induced gastric ulceration[38]Aspirin-induced gastric ulceration[168,169]Intestine Intestinal I/R injury[170,171]Ulcerative colitis[172,173]Colon inflammation[174]Sepsis-induced intestinal injury[87]Necrotizing enterocolitis[175]Liver Liver I/R injury[71,98,[176][177][178]Chronic hepatitis B[179]Nonalcoholic steatohepatitis[180]Liver injury induced by massive hepatectomy[67,93,181]Liver injury induced by obstructive jaundice[31]Liver injury induced by endotoxin[35]Liver injury induced by acetaminophen[47]Liver injury induced by carbon tetrachloride[42]Liver injury induced by concanavalin A[182]Liver cirrhosis[183]Liver fibrosis[184]Pancreas ...
Article
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Therapeutic effects of molecular hydrogen for a wide range of disease models and human diseases have been investigated since 2007. A total of 321 original articles have been published from 2007 to June 2015. Most studies have been conducted in Japan, China, and the USA. About three-quarters of the articles show the effects in mice and rats. The number of clinical trials is increasing every year. In most diseases, the effect of hydrogen has been reported with hydrogen water or hydrogen gas, which was followed by confirmation of the effect with hydrogen-rich saline. Hydrogen water is mostly given ad libitum. Hydrogen gas of less than 4 % is given by inhalation. The effects have been reported in essentially all organs covering 31 disease categories that can be subdivided into 166 disease models, human diseases, treatment-associated pathologies, and pathophysiological conditions of plants with a predominance of oxidative stress-mediated diseases and inflammatory diseases. Specific extinctions of hydroxyl radical and peroxynitrite were initially presented, but the radical-scavenging effect of hydrogen cannot be held solely accountable for its drastic effects. We and others have shown that the effects can be mediated by modulating activities and expressions of various molecules such as Lyn, ERK, p38, JNK, ASK1, Akt, GTP-Rac1, iNOS, Nox1, NF-κB p65, IκBα, STAT3, NFATc1, c-Fos, and ghrelin. Master regulator(s) that drive these modifications, however, remain to be elucidated and are currently being extensively investigated.
... In 2001, studies demonstrated that high-pressure molecular hydrogen gas has anti-inflammatory properties that can cure parasite-induced animal liver inflammation (Gharib et al. 2001). Subsequent cellular and animal studies as well as clinical experiments in diverse biomedical fields have demonstrated the preventive and therapeutic effects of molecular hydrogen in various organs, including the brain, heart, lung, pancreas, and liver, through its antioxidative stress, anti-inflammatory, antiapoptotic, and various other biological effects (Qiu et al. 2011;Hou et al. 2012;Xie et al. 2012;Luo et al. 2015;Liu et al. 2020b). The action of molecular hydrogen in the body is moderate and no side effects have been identified so far. ...
Article
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Acute lung injury (ALI) and acute respiratory distress syndrome, which is a more severe form of ALI, are life-threatening clinical syndromes observed in critically ill patients. Treatment methods to alleviate the pathogenesis of ALI have improved to a great extent at present. Although the efficacy of these therapies is limited, their relevance has increased remarkably with the ongoing pandemic caused by the novel coronavirus disease 2019 (COVID-19), which causes severe respiratory distress syndrome. Several studies have demonstrated the preventive and therapeutic effects of molecular hydrogen in the various diseases. The biological effects of molecular hydrogen mainly involve anti-inflammation, antioxidation, and autophagy and cell death modulation. This review focuses on the potential therapeutic effects of molecular hydrogen on ALI and its underlying mechanisms and aims to provide a theoretical basis for the clinical treatment of ALI and COVID-19.
... Zhang et al. indicated that matrine (one of the main active components of Chinese herb Sophora flavescens Ait) exhibits a protective effect on LPSinduced ALI by inhibiting of the inflammatory responses, which may involve the suppression of ROS and tissue oxidative stress [40]. In addition, Qiu et al. also demonstrated that hydrogen inhalation ameliorates LPS-induced ALI and it may exert its protective role by preventing the activation of ROS-JNK-caspase-3 pathway [41]. Recently, phagocytic NOX-ROS signaling has been shown to play a critical role in promoting TNF-a-induced, NF-kBdependent acute inflammatory responses and tissue injury specifically in the lungs, which is effected by preferential leukocyte infiltration [42]. ...
... Moreover, it was found that H 2 downregulated the expression levels of p-JNK1/2 and p-AKT in the vmPFC. Previous studies also suggested that inhibition of JNK1/2 and AKT activations may be involved in the anti-apoptotic ability of H 2 (55,56). Thus, besides JNK1/2 and AKT signaling, inhibition of p38 MAPK signaling is also a plausible mechanism for H 2 -dependent neuroprotection following SAH. ...
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Subarachnoid hemorrhage (SAH) results in high rates of mortality and lasting disability. Hydrogen gas (H2) is an antioxidant with demonstrated neuroprotective efficacy. The present study examined the therapeutic efficacy of H2 inhalation on early brain injury following experimental SAH in rats and the potential underlying molecular mechanisms. The rats were randomly separated into three groups (n=36 per group): Sham, SAH and SAH + H2. Endovascular perforation of the right internal carotid artery was used to establish SAH. After perforation, rats in the SAH + H2 group inhaled 2.9% H2 with regular oxygen for 2 h. Then, 24 h post-SAH, TUNEL staining was used to detect apoptotic neurons, and both immunostaining and western blotting were conducted to examine changes in p38 MAPK activity and the expression levels of apoptotic regulators (Bcl-2, Bax and cleaved caspase-3) in the ventromedial prefrontal cortex. Then, 30 day post-SAH, Nissl staining was performed to detect neuronal injury, brain MRI was conducted to detect gross changes in brain structure and metabolism, the open field test was used to assess anxiety and the novel object recognition test was performed to assess memory. H2 inhalation following experimental SAH stabilized brain metabolites, improved recognition memory and reduced anxiety-like behavior, the neuronal apoptosis rate, phosphorylated p38 MAPK expression, cleaved caspase-3 expression and the Bax/Bcl-2 ratio. Collectively, the present results suggested that H2 inhalation can alleviate SAH-induced cognitive impairment, behavioral abnormalities and neuronal apoptosis in rats, possibly via inhibition of the p38 MAPK signal pathway.
... Lung edema derived from the disruption of the capillary-alveolar barrier is an important characteristic of ALI [31]. The lung wet/dry ratio is an indicator of pulmonary edema [29], and the total protein concentrations in the BLAF serve as a measurement of pulmonary vascular permeability [32]. Our results demonstrated that LPS stimulation led to an increased lung wet/dry ratio and elevated inflammatory cell count in the BALF. ...
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Acute lung injury (ALI) is a serious respiratory syndrome characterized with uncontrolled inflammatory response. Oxyberberine has strong potential for clinical usage since it showed strong anti-inflammatory, antifungal, and antiarrhythmic effects in various diseases. In the present study, we evaluated whether oxyberberine can inhibit lipopolysaccharide- (LPS-) induced ALI in vivo and further evaluated the possible involvement of mitophagy in vitro by using A549 cells, a human lung epithelial cell line. Our in vivo study shows that oxyberberine significantly inhibited LPS-induced lung pathological injury and lung edema, as indicated by the changes in lung wet/dry ratio and total protein levels in the BALF in mice. Moreover, oxyberberine inhibited inflammation, as indicated by the changes of neutrophil accumulation and production of proinflammatory cytokines including tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 in both the lung and bronchoalveolar lavage fluid (BALF) in ALI mice. Our in vitro study shows that LPS significantly decreased the protein level of mitochondrial proteins, including cytochrome c oxidase subunit IV (COX IV), p62, and mitofusin-2 (Mfn2) in A549 cells. In addition, LPS induced significant Parkin1 translocation from cytoplasm to mitochondria. These changes were significantly inhibited by oxyberberine. Notably, the inhibitory effect of oxyberberine was almost totally lost in the presence of lysosome fusion inhibitor bafilomycin A1 (Baf), a mitophagy inhibitor. In conclusion, the present study demonstrated that oxyberberine alleviated LPS-induced inflammation in ALI via inhibition of Parkin-mediated mitophagy.
... There have been many reports that breathing hydrogen can effectively assist in the treatment of a variety of lung diseases, including but not limited to exogenous infection, endogenous immune dysfunction, cellular dysfunction and even COVID-19 [18][19][20][21]. As early as 2011, some researchers found that hydrogen can effectively relieve the acute lung injury caused by LPS stimulation, and can significantly reduce the expression of various inflammatory factors [22]. Meanwhile, in 2020, it was found that the single inhalation of hydrogen with a low concentration for 45 min was quite effective in modulating airway inflammation in patients with asthma and COPD [23]. ...
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Hydrogen therapy has recently attracted increasing attention as an emerging and promising therapeutic technology due to its selective antioxidant properties and regulatory capacities in vivo. To effectively solve the low solubility issue of hydrogen, a variety of nanomaterials and devices for hydrogen supply have recently been developed, aiming to increase the concentration of hydrogen in specific disease location and realize controlled hydrogen release and combined treatment. In this review, we mainly focus on the latest advances in using hydrogen-generating devices and nanomaterials for hydrogen therapy. These developments include the sustained release of H2 , controlled release of H2 , and versatile modalities of synergistic therapy, etc. Also, bio-safety issues and challenges are discussed to further promote the clinical applications of hydrogen therapy and the development of hydrogen medicine.
... H 2 may prevent necrosis by maintaining ATP levels (178). Regarding apoptosis, hydrogen downregulates caspase-8 (182), which is a key protein that initiates the extrinsic pathway (183), and it also increases the Blc2/Bax ratio (67,184), which are antiapoptotic proteins that inhibit apoptosis by stabilizing the outer mitochondrial membrane and prevent the release of cytochrome c into the cytosol through the intrinsic pathway (185). ...
Article
There are many situations of excessive production of reactive oxygen species (ROS) such as radiation, ischemia/reperfusion (I/R), and inflammation. ROS contribute to and arises from numerous cellular pathologies, diseases, and aging. ROS can cause direct deleterious effects by damaging proteins, lipids, and nucleic acids as well as exert detrimental effects on several cell signaling pathways. However, ROS are important in many cellular functions. The injurious effect of excessive ROS can hypothetically be mitigated by exogenous antioxidants, but clinically this intervention is often not favorable. In contrast, molecular hydrogen provides a variety of advantages for mitigating oxidative stress due to its unique physical and chemical properties. H2 may be superior to conventional antioxidants, since it can selectively reduce ●OH radicals while preserving important ROS that are otherwise used for normal cellular signaling. Additionally, H2 exerts many biological effects, including anti-oxidation, anti-inflammation, anti-apoptosis, and anti-shock. H2 accomplishes these effects by indirectly regulating signal transduction and gene expression, each of which involve multiple signaling pathways and crosstalk. The Keap1-Nrf2-ARE signaling pathway, which can be activated by H2 , plays a critical role in regulating cellular redox balance, metabolism, and inducing adaptive responses against cellular stress. H2 also influences the crosstalk among the regulatory mechanisms of autophagy and apoptosis, which involve MAPKs, p53, Nrf2, NF-κB, p38 MAPK, mTOR, etc. The pleiotropic effects of molecular hydrogen on various proteins, molecules and signaling pathways can at least partly explain its almost universal pluripotent therapeutic potential.
... 22 hydrogen could alleviate acute lung injury by reducing IL-6 and TNF-α expression. 24 Liu et al proposed that hydrogen treatment might become a new and effective method for COPD, 25 in cutaneous ischemia/reperfusion injury in a mouse model of pressure ulcer. 30 Our study also found the MCP1 level in peripheral blood significantly decreased after hydrogen inhalation, that hydrogen inhalation may produce protection against smoking. ...
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Background: Hydrogen was proven to have anti-oxidative and anti-inflammation effects to various diseases. Aim: We wish to investigate the acute effects of inhaled hydrogen on airway inflammation in patients with asthma and chronic obstructive pulmonary disease (COPD). Design: Prospective study. Methods: 2.4% hydrogen containing steam mixed gas (XEN) was inhaled once for 45 minutes in 10 patients with asthma and 10 patients with COPD. The levels of granulocyte-macrophage colony stimulating factor (GM-CSF), interferon-γ (IFNγ), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6), et al. in peripheral blood and exhaled breath condensate (EBC) before and after "XEN" inhalation were measured. Results: 45 minutes "XEN" inhalation once decreased monocyte chemotactic protein 1 (MCP1) level in both COPD (564.70 to 451.51pg/mL, P = 0.019) and asthma (386.39 to 332.76 pg/mL, P = 0.033) group, while decreased IL-8 level only in asthma group (5.25 to 4.49pg/mL, P = 0.023). The level of EBC soluble cluster of differentiation-40 ligand (sCD40L) in COPD group increased after inhalation (1.07 to 1.16pg/mL, P = 0.031), while IL-4 and IL-6 levels in EBC were significantly lower after inhalation in the COPD (0.80 to 0.64pg/mL, P = 0.025) and asthma (0.06 to 0.05pg/mL, P = 0.007) group respectively. Conclusions: A single inhalation of hydrogen for 45 minitues attenuated inflammatory status in airways in patients with asthma and COPD.
... However, this damage was reversed by biochanins A. Lung edema derived from the disruption of the capillary-alveolar barrier is an important characteristic of ALI [15]. Lung wet/dry ratio is an indicator of pulmonary edema [16], and the total protein concentrations in the BLAF serve as a measurement of pulmonary vascular permeability [17]. Our results demonstrated LPS stimulation led to increased lung wet/dry ratio, and elevated inflammatory cell count in the BALF. ...
Article
Acute lung injury (ALI) is a serious respiratory syndrome featured with uncontrolled inflammatory response. Biochanin A has been showed to possess and anti-inflammatory effect. This study intended to explore the suppression of biochanin A on lipopolysaccharide (LPS)-induced ALI in mice. Seven hours later LPS-induced ALI model established, the indexes including, pathological changes, MPO activity, wet/dry ratio, proinflammatory cytokines TNF-α, IL-1β, and IL-6, production, as well as and TLR4/NF-κB and PPAR-γ signaling pathway expression were compared bwtween different groups. In addition, bronchoalveolar lavage fluid (BALF) was collected and the levels of total protein, inflammatory cells and TNF-α, IL-1β, and IL-6 were detected. The results revealed that LPS lead to significantly lung pathological injury, and damage of lung vascular permeability showing by higher lung wet/dry ratio and total protein levels in the BALF when compared to the control group mice. However, these changes significantly reversed by biochanin A. Moreover, the levels of inflammatory cells in BALF, proinflammatory cytokines TNF-α, IL-1β, and IL-6, in both lung and BALF were also dose-dependently reduced by biochanin A during ALI process. To investigate the anti-inflammatory mechanisms of biochanin A, we found that biochanin A significantly inhibited the activation of TLR4/NF-κB signaling pathway induced by LPS. Furthermore, the expression of PPAR-γ also markedly increased in the mice after treated with biochanin A. In conclusion, biochanin A alleviated LPS-induced ALI by inhibiting the inflammatory response, which was mediated via down-regulating the activation of TLR4/NF-κB signaling pathway and enhancing the expression of PPAR-γ.
... In addition, immunohistochemical staining revealed that the increase in MOMA-2-positive macrophages significantly correlated with the numbers of both p16 INK4a -and p21-ECs in the same aortic region (Fig. 5). Several studies reported that H 2 administration attenuated cellular inflammation and production of inflammatory cytokines in experimental animal models of inflammatory diseases such as zymosan-induced inflammation, inflammatory bowel disease, and lipopolysaccharide (LPS)-induced inflammation [44][45][46] . H 2 inhibited LPS-induced phosphorylation of apoptosis signal-related kinase 1 (ASK1) via Toll-like receptors (TLRs) 47 . ...
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The main cause of arteriosclerosis is atherosclerosis in the aorta. Atherosclerosis is recognized as a chronic inflammatory condition that begins with the dysfunction or activation of arterial endothelium. Low-density lipoprotein (LDL) and especially its oxidized form play a key role in endothelial dysfunction and atherogenesis. Recent studies showed that senescent cells are involved in the development and progression of atherosclerosis, and eliminating senescent cells suppresses the senescence-associated secretory phenotype. We previously reported that molecular hydrogen-rich water (HW) has antioxidant and anti-inflammatory effects in numerous diseases. Here, we used LDL receptor-deficient mice fed a high-fat diet (HFD) for 13 weeks as a model for atherosclerosis and evaluated the effects of continuous administration of HW. The numbers of endothelial cells in the atheroma expressing the senescence factors p16INK4a and p21 decreased in HFD-fed mice given HW compared with HFD-fed mice given control water. Furthermore, macrophage infiltration and Tnfα expression in the atheroma were also suppressed. These results suggest that vascular aging can be suppressed by HW.
... reacts with the hydroxyl radical (·OH) to produce water in the body. Second, H 2 can easily penetrate membranes and diffuse into the cytosol and nucleus, making it highly effective in reducing cytotoxic substances (45). Third, there is no influence of H 2 on the physiologic properties (24,46). ...
Article
Background: Chronic obstructive pulmonary disease (COPD) is a chronic lung disease with limited treatment options. Hydrogen (H2) has been shown to be anti-oxidative and anti-inflammatory. This study aimed to evaluate the beneficial effects of H2 inhalation on COPD development in mice. Methods: A COPD mouse model was established in male C57BL mice by cigarette smoke (CS) exposure. The H2 intervention was administered by atomisation inhalation. Lung functions were assessed by using Buxco lung function measurement system. The inflammatory cells were counted and the levels of IL-6 and KC in BALF were assayed with ELISA. The lung tissue was subjected to H&E or PAS or Masson's trichrome stain. Furthermore, 16HBE cells were used to evaluate the effects of H2 on signaling change caused by hydrogen peroxide (H2O2). H2O2 was used to treat 16HBE cells with or without H2 pretreatment. The IL-6 and IL-8 levels in cell culture medium were measured. The levels of phosphorylated ERK1/2 and nucleic NF-κB in lungs and 16HBE cells were determined. Results: H2 ameliorated CS-induced lung function decline, emphysema, inflammatory cell infiltration, small-airway remodelling, goblet-cell hyperplasia in tracheal epithelium and activated ERK1/2 and NF-κB in mouse lung. In 16HBE airway cells, H2O2 increased IL-6 and IL-8 secretion in conjunction with ERK1/2 and NF-κB activation. These changes were reduced by H2 treatment. Conclusions: These findings demonstrated that H2 inhalation could inhibit CS-induced COPD development in mice, which is associated with reduced ERK1/2 and NF-κB-dependent inflammatory responses.
... Liu et al. [6] reported that, combined early fluid resuscitation and hydrogen inhalation attenuated lung and intestine injury. Additionally, Qiu et al. [40] found that hydrogen inhalation could ameliorate lipopolysaccharide-induced acute lung injury in mice. These previous studies all suggested that H 2 had effective treatment effect for lung injury, and our study, for the first time, clarified the function of H 2 on lung cancer. ...
Article
Lung cancer is one of the most common lethal malignancies in the globe. The patients' prognoses are dim due to its high metastatic potential and drug resistance. Therefore, in the present study, we aim to find a more potent therapeutic approach for lung cancer. We mainly explored the function of hydrogen gas (H2) on cell viability, apoptosis, migration and invasion in lung cancer cell lines A549 and H1975 by CCK-8, flow cytometry, wound healing and transwell assays, respectively. We used RNA-seq, qPCR and western blotting to detect the different expression genes (DEGs) between H2 group and control group to find the gene related to chromosome condensation. Besides, we confirmed the structural maintenance of chromosomes 3 (SMC3) and H2 on the progression of lung cancer in vitro and vivo. Results showed that H2 inhibited cell viability, migration and invasion, and catalyzed cell apoptosis and H2 induced A549 and H1975 cells G2/M arrest. Besides, H2 down-regulated the expression of NIBPL, SMC3, SMC5 and SMC6, and also reduced the expression of Cyclin D1, CDK4 and CDK6. H2 translocated the subcellular location of SMC3 during cell division and decreased its stability and increased its ubiquitination in both A549 and H1975 cells. In addition, inhibition of the proliferation, migration and invasion and promotion of the apoptosis of A549 and H1975 cells induced by H2 were all abolished when overexpressed SMC3 in the presence of H2. Animal experimental assay demonstrated that the tumor weight in H2 group was significantly smaller than that in control group, but was bigger than cis-platinum group. The expression of Ki-67, VEGF and SMC3 were decreased when mice were treated with H2 or cis-platinum, especially for cis-platinum. All data suggested that H2 inhibited lung cancer progression through down-regulating SMC3, a regulator for chromosome condensation, which provided a new method for the treatment of lung cancer.
... Bcl-xl is an anti-apoptotic gene that exerts anti-apoptosis effects by inhibiting the activation of caspase proteases (30). Proapoptotic genes are dominant during early apoptosis, during which Bcl-xl expression is decreased, and as apoptosis develops, the internal anti-apoptotic mechanism is activated (31). The present study demonstrated that allicin significantly increased the expression of the Bcl-2 anti-apoptotic protein and inhibited caspase-3/-9 activity in ALI model neonatal rats, which may occur via the PI3K/Akt pathway. ...
Article
Allicin is an oxygenated carotenoid derivative that exhibits strong antioxidant activity, which effectively removes reactive oxygen species from the body and has important roles in disease prevention and treatment. Therefore, the present study aimed to investigate whether allicin attenuates lipopolysaccharide (LPS)‑induced acute lung injury (ALI) in neonatal rats and the potential underlying mechanisms. An LPS‑induced ALI neonatal rat model was utilized to assess the therapeutic value and mechanisms of allicin. Following allicin treatment, increases in lung wet/dry ratio and the lung protein concentration were significantly suppressed in LPS‑induced ALI neonatal rats. Furthermore, ELISA results demonstrated that allicin significantly reduced the levels of malondialdehyde, tumor necrosis factor‑α and interleukin‑6, and increased superoxide dismutase activity, in the bronchoalveolar lavage fluid of LPS‑treated rats. Additionally, allicin administration increased the protein expression of Bcl‑2 and reduced the activity of caspase‑3/-9, as determined by western blotting or ELISA, respectively, and increased phosphatidylinositol 3‑kinase (PI3K) and phosphorylated‑Akt protein levels, in LPS‑treated ALI neonatal rats. The results of the present study indicate that allicin attenuate LPS‑induced ALI in neonatal rats by ameliorating oxidative stress, inflammation and apoptosis via the PI3K/Akt pathway. Allicin may be used for development of a novel drug for treatment of ALI.
... Qiu et al found that 2% hydrogen could alleviate acute lung injury by reducing TNF-α and IL-6 expression. 36 This study showed that hydrogen could reduce the number of inflammatory cells in the BALF and reduce the expression of TNF-α, IL-6, IL-17, and IL-23; in this respect, 22% and 41.6% hydrogen yielded better outcome than 2% hydrogen. These results suggest that hydrogen inhalation alleviated inflammation through its inhibitory actions on the number of inflammatory cells, the expression of inflammatory factors, or both. ...
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Background Chronic obstructive pulmonary disease (COPD) is a progressive pulmonary disease caused by harmful gases or particles. Recent studies have shown that 2% hydrogen or hydrogen water is effective in the treatment and prevention of a variety of diseases. This study investigated the beneficial effects and the possible mechanisms of different hydrogen concentrations on COPD. Methods A rat COPD model was established through smoke exposure methods, and inhalation of different concentrations of hydrogen was used as the intervention. The daily condition of rats and the weight changes were observed; lung function and right ventricular hypertrophy index were assessed. Also, white blood cells were assessed in bronchoalveolar lavage fluid. Pathologic changes in the lung tissue were analyzed using light microscopy and electron microscopy; cardiovascular structure and pulmonary arterial pressure changes in rats were observed using ultrasonography. Tumor necrosis factor alpha, interleukin (IL)-6, IL-17, IL-23, matrix metalloproteinase-12, tissue inhibitor of metalloproteinase-1, caspase-3, caspase-8 protein, and mRNA levels in the lung tissue were determined using immunohistochemistry, Western blot, and real-time polymerase chain reaction. Results The results showed that hydrogen inhalation significantly reduced the number of inflammatory cells in the bronchoalveolar lavage fluid, and the mRNA and protein expression levels of tumor necrosis factor alpha, IL-6, IL-17, IL-23, matrix metalloproteinase-12, caspase-3, and caspase-8, but increased the tissue inhibitor of metalloproteinase-1 expression. Furthermore, hydrogen inhalation ameliorated lung pathology, lung function, and cardiovascular function and reduced the right ventricular hypertrophy index. Inhalation of 22% and 41.6% hydrogen showed better outcome than inhalation of 2% hydrogen. Conclusion These results suggest that hydrogen inhalation slows the development of COPD-like lung disease in a cigarette smoke-induced rat model. Higher concentrations of hydrogen may represent a more effective way for the rat model.
... Results are mean ± SD (n = 6/group) LPS is one of the known culprits in the production of reactive oxygen species and pro-inflammatory cytokines including TNF-α, IL-1β, and so on (Kim et al., 2014). Production of these cytokines contributes to acquisition of neutrophils in the lung, leading to ALI (Bhatia & Moochhala, 2004;Qiu et al., 2011). In our study, we found LPS-induced the production of pro-inflammatory factors TNF-α and IL-1β, inhibiting autophagy with CLQ further promoted the production of these cytokines. ...
Article
Understanding the role and underlying regulation mechanism of autophagy in lipopolysaccharide-induced lung injury (LPS-LI) may provide potentially new pharmacological targets for treatment of acute lung injury. The aim of this study was to investigate the functional significance of autophagy in LPS-LI. The autophagy of human pulmonary microvascular endothelial cells (HPMVECs) and mice was inhibited before they were challenged with LPS. In vitro, permeability, vitality and the lactate dehydrogenase (LDH) release rate of the cells were detected, the zonula occluden-1 (ZO-1) expression and the stress fiber formation were determined. In vivo, the lung injury was assessed. We found LPS caused high permeability and increased LDH release rate, lowered viability of the cells, inhibited the ZO-1 expression and induced stress fiber formation, these effects were further aggravated by prohibiting the level of autophagy. Consistently, in in vivo experiments, LPS induced serious lung injury, which was reflected as edema, leukocyte infiltration and hemorrhage in lung tissue, and the high concentration of pro-inflammation cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-1β in bronchoalveolar lavage fluid (BALF). Inhibiting autophagy further exacerbated LPS-LI. It appears that autophagy played a protective role in LPS-LI in part through restricting the injury of lung microvascular barrier. This article is protected by copyright. All rights reserved.
... Cytokines, such as TNF-α and interleukins, are commonly observed at inflammatory sites and can increase the inflammatory response by initiating the infiltration of innate immune cells such as neutrophils and macrophages into inflamed tissues. 14 Myeloperoxidase (MPO) has been used as a neutrophil cell marker, 15,16 and the activity of this enzyme can be used to determine neutrophil infiltration. Tight junctions in the stratum granulosum confer the mechanical strength of the rumen epithelium. ...
Article
To investigate the effect of sodium butyrate on high-concentratediet-induced local inflammation of the rumen epithelium, 18 mid-lactating dairy goats were randomly assigned to 3 groups: a low-concentrate diet group as the control (concentrate: forage=4:6), a high-concentrate (HC) diet group (concentrate: forage=6:4), and a sodium butyrate (SB) group (concentrate: forage=6:4, with 1% SB by weight). The results showed that, with the addition of sodium butyrate, the concentration of lipopolysaccharide(LPS) in rumen fluid (2.62×10(4)±2.90×10(3) EU/mL) was significantly lower than that in the HC group (4.03×10(4)±2.77×10(3) EU/mL). The protein abundance of pp65, gene expression of pro-inflammatory cytokines, and activity of myeloperoxidase (MPO) and matrix metalloproteinase (MMP)-2,9 in the rumen epithelium were significantly down-regulated by SB compared with those in the HC group. With sodium butyrate administration, the concentration of NH3-N (19.2±0.890 mM) in the rumen fluid was significantly higher than that for the HC group (12.7±1.38 mM). Severe disruption of the rumen epithelium induced by HC was also ameliorated by dietary SB. Therefore, local inflammation and disruption of the rumen epithelium induced by HC were alleviated with SB administration.
... 2,3 Since the first report of the marked effect of molecular hydrogen on a rat model of cerebral infarction, the beneficial effects of hydrogen have been reported in more than 60 model animals and more than 10 human diseases, especially in oxidative-stress mediated and inflammatory diseases. 4 The therapeutic effects of hydrogen have been reported in lung injuries due to ischemia/reperfusion 5,6 ; oxygen 7,8 ; ventilation 9,10 ; LPS [11][12][13][14] ; paraquat 15,16 ; smoke 17 ; and cigarette smoking. 18 The effects of hydrogen have also been reported in lung injuries secondary to intestinal ischemia/reperfusion 19 and extensive skin burn. ...
Article
Bronchopulmonary dysplasia (BPD) is characterized by developmental arrest of the alveolar tissue. Oxidative stress is causally associated with development of BPD. The effects of hydrogen have been reported in a wide range of disease models and human diseases especially caused by oxidative stress. We made a rat model of BPD by injecting lipopolysaccharide (LPS) into the amniotic fluid at E16.5. The mother started drinking hydrogen-rich water from E9.5 and also while feeding milk. Hydrogen normalized LPS-induced abnormal enlargement of alveoli at P7 and P14. LPS increased staining for nitrotyrosine and 8-OHdG of the lungs, and hydrogen attenuated the staining. At P1, LPS treatment decreased expressions of genes for FGFR4, VEGFR2, and HO-1 in the lungs, and hydrogen increased expressions of these genes. In contrast, LPS treatment and hydrogen treatment had no essential effect on the expression of SOD1. Inflammatory marker proteins of TNFα and IL-6 were increased by LPS treatment, and hydrogen suppressed them. Treatment of A549 human lung adenocarcinoma epithelial cells with 10% hydrogen gas for 24 hr decreased production of reactive oxygen species in both LPS-treated and untreated cells. Lack of any known adverse effects of hydrogen makes hydrogen a promising therapeutic modality for BPD. Pediatr Pulmonol. © 2016 Wiley Periodicals, Inc.
... In contrast, Itoh and colleagues regarded H 2 as a gaseous signalling molecule due to the result showing that H 2 could attenuate the phosphorylation of FcεRI-associated Lyn as well as its downstream signal transduction (such as JNK, p38 MAPK, ERK) in rat RBL-2H3 mast cells, followed by the inhibition of NADPH oxidase activity and reduction of hydrogen peroxide (H 2 O 2 ) levels [76]. In addition to ROS activation, JNK can be activated by the LPS-induced inflammatory response and attenuated by hydrogen inhalation [77]. The data, obtained from human histocytic lymphoma U937 cells, indicated that H 2 O 2 and ·OH could activate PI3K-Akt and PLC-Ras-Raf-ERK signaling pathways [78], whereas Xu and Zhang discovered that saturated HS decreased LPS-induced ERK phosphorylation in a rat model of acute liver dysfunction [79]. ...
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Early acute kidney injury (AKI) in severely burned patients predicts a high mortality that is multi-factorial. Hydrogen has been reported to alleviate organ injury via selective quenching of reactive oxygen species. This study investigated the potential protective effects of hydrogen against severe burn-induced early AKI in rats. Severe burn were induced via immersing the shaved back of rats into a 100°C bath for 15 s. Fifty-six Sprague-Dawley rats were randomly divided into Sham, Burn + saline, and Burn + hydrogen-rich saline (HS) groups, and renal function and the apoptotic index were measured. Kidney histopathology and immunofluorescence staining, quantitative real-time PCR, ELISA and western blotting were performed on the sera or renal tissues of burned rats to explore the underlying effects and mechanisms at varying time points post burn. Renal function and tubular apoptosis were improved by HS treatment. In addition, the oxidation-reduction potential and malondialdehyde levels were markedly reduced with HS treatment, whereas endogenous antioxidant enzyme activities were significantly increased. HS also decreased the myeloperoxidase levels and influenced the release of inflammatory mediators in the sera and renal tissues of the burned rats. The regulatory effects of HS included the inhibition of p38, JNK, ERK and NF-κB activation, and an increase in Akt phosphorylation. Hydrogen can attenuate severe burn-induced early AKI; the mechanisms of protection include the inhibition of oxidative stress induced apoptosis and inflammation, which may be mediated by regulation of the MAPKs, Akt and NF-κB signalling pathways.
... Studies have shown that hydrogen inhalation can markedly prevent the lipopolysaccharide (LPS)-induced apoptosis, which blocked the activation of JNK, downregulated the level of Bax, caspase-3 and upregulated the Bcl-xl expression. 39 Inhalation hydrogen in the early stage of Ventilator Induced Lung Injury (VILI) can activate NF-kB, correlated with the elevated level of Bcl-2, which was inhibited by SN50, an inhibitor of NF-kB. 40 Finally the decreased Bax/Bcl-2 ratio inhibited the expression of capase-3 and apoptosis. ...
Article
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Many pathways have been reported involving the effect of hydrogen-rich saline on protecting skin flap partial necrosis induced by the inflammation of ischemia/reperfusion injury. This study focused on the influence of hydrogen-rich saline treatment on apoptosis pathway of ASK-1/JNK and Bcl-2/Bax radio in I/R injury of skin flaps. Adult male Sprague-Dawley rats were divided into three groups. Group 1 was sham surgery group, Group 2 and 3 were ischemia/reperfusion surgery treated with physiological saline and hydrogen-rich saline respectively. Blood perfusion of flap was measured by Laser doppler flowmeters. Hematoxylin and eosin staining was used to observe morphological changes. Early apoptosis in skin flap was observed through TUNEL staining and presented as the percentage of TUNEL-positive cells of total cells. pASK-1, pJNK, Bcl-2 and Bax were examined by immunodetection. In addition Bcl-2, Bax and caspase-3 were detected by qPCR. Caspase-3 activity was also measured. Compared to the Group 2, tissues from the group 3 were observed with a high expression of Bcl-2 and a low expression of pASK-1, pJNK, and Bax, a larger survival area and a high level of blood perfusion. Hydrogen-rich saline ameliorated inflammatory infiltration and decreased cell apoptosis. The results indicate that hydrogen-rich saline could ameliorate ischemia/reperfusion injury and improve flap survival rate by inhibiting the apoptosis factor and, at the same time, promoting the expression of anti-apoptosis factor. Copyright © 2015. Published by Elsevier Ltd.
... Endotoxin is thought to be the most important pathogen that leads to the development of ALI [4,5]. Excessive cytokinemediated inflammation plays a fundamental role in the pathogenesis of ALI [6,7]. Increased levels of local and systemic inflammatory mediators, such as tumor necrosis factor (TNF)-α and interleukin (IL)-1β and IL-6, and activated leukocytes, lead to systemic inflammatory response syndrome (SIRS) [8,9]. ...
Article
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Anti-inflammatory and anti-apoptotic effects of polydatin (PD) have been demonstrated in our previous studies. Recently, we have found that PD treatment can ameliorate burn-induced acute lung injury (ALI). In the present study, we hypothesized that PD may provide protective effect against LPS-induced ALI through reducing inflammation and apoptosis. Rats were respectively pretreated with PD at doses of 15, 30 and 45 mg/kg weight, followed by intratracheal administration of lipopolysaccharide (LPS). LPS-challenged rats exhibited significant lung injury characterized by the deterioration of histopathology, pulmonary microvascular hyperpermeability, wet-to-dry weight ratio, and oxygenation index, which was attenuated by PD (30 and 45 mg/kg) treatment. Moreover, PD (30 and 45 mg/kg) treatment inhibited LPS-induced inflammatory response, as evidenced by the downregulation of lung myeloperoxidase activity, total cells and PMNs in bronchoalveolar lavage fluid, and the systemic levels of the pro-inflammatory cytokines. Furthermore, PD (30 and 45 mg/kg) treatment remarkably improved LPS-induced increase in TUNEL (deoxynucleotidyl transferase dUTP nick end labeling) staining-positive cells, caspase 3 activity, Bax over-expression and Bcl-2 down-expression. In conclusion, these results demonstrate that PD (30 and 45 mg/kg) treatment attenuates LPS-induced ALI through reducing lung inflammation and apoptosis.
... ALI is most often seen as part of a systemic inflammatory process where the lung manifestations parallel those of other tissueswidespread destruction of the capillary endothelium, extravascation of protein rich fluid and interstitial edema [2]. Gram-negative bacterial infections are the main cause of ALI, and lipopolyssacharide (LPS), a major component of the cell wall of Gram-negative bacteria, is known to be a culprit for the production of reactive oxygen species (ROS) and pro-inflammatory cytokines, resulting in increased infiltration of inflammatory cells [3,4]. The binding of LPS to TLR4 activates the nuclear factor (NF)-κB so to enhance the production of key pro-inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, and monocyte chemotactic protein-1 (MCP-1) [5,6]. ...
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The fruit hull of Gleditsia sinensis (FGS) used in traditional Asian medicine was reported to have a preventive effect on lung inflammation in an acute lung injury (ALI) mouse model. Here, we explored FGS as a possible therapeutics against inflammatory lung diseases including ALI, and examined an underlying mechanism for the effect of FGS. The decoction of FGS in water was prepared and fingerprinted. Mice received an intra-tracheal (i.t.) FGS 2 h after an intra-peritoneal (i.p.) injection of lipopolysaccharide (LPS). The effect of FGS on lung inflammation was determined by chest imaging of NF-κB reporter mice, counting inflammatory cells in bronchoalveolar lavage fluid, analyzing lung histology, and performing semi-quantitative RT-PCR analysis of lung tissue. Impact of Nrf2 on FGS effect was assessed by comparing Nrf2 knockout (KO) and wild type (WT) mice that were treated similarly. Bioluminescence from the chest of the reporter mice was progressively increased to a peak at 16 h after an i.p. LPS treatment. FGS treatment 2 h after LPS reduced the bioluminescence and the expression of pro-inflammatory cytokine genes in the lung. While suppressing the infiltration of inflammatory cells to the lungs of WT mice, FGS post-treatment failed to reduce lung inflammation in Nrf2 KO mice. FGS activated Nrf2 and induced Nrf2-dependent gene expression in mouse lung. FGS post-treatment suppressed lung inflammation in an LPS-induced ALI mouse model, which was mediated at least in part by Nrf2. Our results suggest a therapeutic potential of FGS on inflammatory lung diseases.
... Numerous studies have reported that up-regulation of inflammatory mediators, such as TNF-␣, IL-1␤, and IL-6, contributes to the risk of the development of acute SIRS, and the inhibition of these factors prevents the development of this syndrome in burn-injury rats. 37,38 Our results in the present study indicate that PD can reduce the elevated levels of local and systemic inflammatory mediators induced by burns. It seems reasonable to speculate that this inhibition might be associated with attenuation of pulmonary pathophysiologic alterations caused by severe burn injury. ...
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Polydatin (PD) has anti-inflammatory and anti-apoptotic effects in ischemic-reperfusion injury. Moreover, inflammatory responses and apoptosis play a role in the development of burn-induced lung injuries. Based on these findings, in this study we investigated the hypothesis that PD can ameliorate lung injury induced by extensive burns via reduction of inflammation and apoptosis. Rats were subjected to 30% total body surface area burn injury followed by resuscitation. The treatment group received 45 mg/kg PD, and the burn group received the same amount of normal saline solution. No burn injury was inflicted in the sham group. Microvascular permeability, interstitial edema, neutrophil recruitment, and histopathological changes were detected by measuring Evans blue concentration, wet-to-dry lung weight ratio (W/D), myeloperoxidase (MPO) activity, and hematoxylin and eosin staining, respectively. To investigate the mechanism of action of PD, enzyme-linked immunosorbent assay, cell counting, terminal deoxyribonucleotidyl transferase-mediated deoxyuridine 5-triphosphate-digoxigenin nick end labeling (TUNEL) staining, fluorometric assay, and Western blot were used for assessing levels of inflammatory cytokines (tumor necrosis factor alpha, interleukin [IL]-1β, and IL-6), total number of cells, and concentration of polymorphonuclear leukocytes (PMNs) in bronchoalveolar lavage fluid (BALF), the number of apoptotic cells, caspase-3 activity, and apoptosis-related proteins including Bax and Bcl-xl, respectively. Burn-injury rats exhibited significant lung injury characterized by the deterioration of histopathological characteristics, pulmonary microvascular hyperpermeability, and a high W/D, which were attenuated by PD (P = .007 for permeability, P = .004 for W/D). PD inhibited the burn-induced inflammatory response, as evidenced by the down-regulation of lung MPO activity (P = .008), total number of cells, PMN concentration in BALF, and the local and systemic levels of the pro-inflammatory cytokines examined. Moreover, PD treatment dramatically prevented burn-induced pulmonary cell apoptosis in lungs, as reflected by the decrease in the number of TUNEL-positive cells (P = .002) and changes in Bax, Bcl-xl, and caspase-3 activity (P = .03). PD ameliorates burn-induced lung injury via its anti-inflammatory and anti-apoptotic effects, and PD treatment may therefore serve as a potential therapeutic target for the treatment of critical burn injuries.
Article
The Golgi apparatus is a crucial component of the endomembrane system and is involved in various physiological and pathological processes. Recently, increasing evidence shows that Golgi participates in oxidative stress in sepsis-induced acute lung injury (ALI). As a kind of reactive oxygen species (ROS), peroxynitrite (ONOO⁻) plays a significant role in a range of oxidative stress-related diseases. Therefore, accurately monitoring the level of ONOO⁻ in the Golgi apparatus is crucial for revealing the mechanism of Golgi ONOO⁻ in the sepsis-induced ALI. Unfortunately, no such probe has been reported to detect Golgi ONOO⁻ in sepsis-induced ALI model. In this work, a Golgi-targetable probe FC-ONOO was designed and synthesized for high sensitivity and selectivity monitoring ONOO⁻ generation during the Golgi stress response in sepsis-induced ALI model. The probe displayed high sensitivity (about 57 nM of detection limit) and good selectivity towards ONOO⁻. Additionally, it was successfully applied in mapping the increase of ONOO⁻ levels in the Golgi of live cells induced by different stimuli. More importantly, the substantial increases in ONOO⁻ levels in the Golgi apparatus were observed in the sepsis-induced ALI of mice for the first time. Given all its outstanding advantages, it has great potential for application in other ONOO⁻ related diseases.
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Nuclear factor erythroid 2-related factor (Nrf2) is a redox-sensitive transcription factor that responds to oxidative stress by activating expressions of key antioxidant and cytoprotective enzymes via the Nrf2-antioxidant response element (ARE) signaling pathway. Our objective was to characterize hyperoxia-induced acute lung injury (HALI) in Nrf2 knock-out (KO) rats to elucidate the role of this pathway in HALI. Adult Nrf2 wildtype (WT), and KO rats were exposed to room air (normoxia) or >95% O2 (hyperoxia) for 48 h, after which selected injury and functional endpoints were measured in vivo and ex vivo. Results demonstrate that the Nrf2-ARE signaling pathway provides some protection against HALI, as reflected by greater hyperoxia-induced histological injury and higher pulmonary endothelial filtration coefficient in KO versus WT rats. We observed larger hyperoxia-induced increases in lung expression of glutathione (GSH) synthetase, 3-nitrotyrosine (index of oxidative stress), and interleukin-1β, and in vivo lung uptake of the GSH-sensitive SPECT biomarker 99mTc-HMPAO in WT compared to KO rats. Hyperoxia also induced increases in lung expression of myeloperoxidase in both WT and KO rats, but with no difference between WT and KO. Hyperoxia had no effect on expression of Bcl-2 (anti-apoptotic protein) or peroxiredoxin-1. These results suggest that the protection offered by the Nrf2-ARE pathway against HALI is in part via its regulation of the GSH redox pathway. To the best of our knowledge, this is the first study to assess the role of the Nrf2-ARE signaling pathway in protection against HALI using a rat Nrf2 knockout model.
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Objective: Chronic obstructive pulmonary disease (COPD) is a respiratory disease with high morbidity and mortality worldwide, so far there is no ideal treatment method. Previous studies have shown that hydrogen (H2) is involved in the treatment of COPD as an antioxidant. In this study, the effect of H2 on M1/M2 polarization of alveolar macrophages in COPD rats was observed, and its anti-inflammatory mechanism was further elucidated. Methods: Twenty-four Sprague-Dawley rats were randomly divided into three groups including the control, COPD and H2 group. A rat model of COPD was established by cigarette exposure combined with lipopolysaccharide (LPS) induction. H2 therapy was administered 2 hours per day for 14 days. Lung function and pathology were assessed. The levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β1 and IL-10 in bronchoalveolar lavage fluid (BALF) and lung tissue were measured by enzyme-linked immunosorbent assay. The mRNA, protein expression and immunoreactivity of inducible nitric oxide synthase (iNOS) and arginase (Arg)-1 in lung were observed by quantitative real-time PCR, western blot and immunohistochemistry. Results: Compared with the control rats, there were a significant decline in lung function, a marked inflammatory infiltration and pulmonary parenchymal remodeling and the increases of IL-6, TNF-α and TGF-β1 levels in BALF and lung tissue, but a lower expression of IL-10 in COPD rats. The iNOS mRNA and protein expression, as well as its optical density (OD), were increased significantly in lung tissue, while those of Arg-1 decreased significantly. H2 treatment improved the lung function and the parenchymal inflammation, reversed the increased levels of IL-6, TNF-α and TGF-β1, and the lower IL-10. Meanwhile, H2 also down-regulated the expression of iNOS, but up-regulated expression of Arg-1 in lung tissue. Conclusion: H2 reduces inflammation in the lung of COPD, which may be related to its inhibition of M1 type polarization and activation of M2 type polarization of alveolar macrophage.
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Aims Limb ischaemia/reperfusion (LIR) occurs in various clinical conditions including critical limb ischaemia, abdominal aortic aneurysm, and traumatic arterial injury. Reperfusion of the acutely ischemic limb can lead to a systemic inflammation response and multiple organ dysfunction syndrome, further resulting in significant morbidity and mortality. Molecular hydrogen exhibits therapeutic activity for the treatment and prevention of many diseases. Our study investigated the possible therapeutic effects of hydrogen and its mechanism of action in a LIR-induced acute lung injury (ALI) model. Materials and methods Limb ischaemia/-reperfusion model was established in mice. The hydrogen-saturated saline was administered by intraperitoneal injection. Protein level of nuclear factor erythroid 2-related factor 2 (Nrf2), haem oxygenase-1 (HO1) and nicotinamide adenine dinucleotide phosphate quinone oxidoreductase 1 (NQO1) was evaluated by immunohistochemistry staining and western blotting. Autophagy-related molecules were evaluated by western blotting. Malondialdehyde (MDA) and superoxide dismutase (SOD) were determined by assay kits. Quantification of ceramides in lung was performed by high-performance liquid chromatography-tandem mass spectrometry. Key findings Molecular hydrogen exhibited a protective effect on the LIR-induced ALI model. Hydrogen decreased malondialdehyde and increased superoxide dismutase activity in lung tissues. Additionally, hydrogen activated Nrf2 signalling in lung tissues. Hydrogen could inhibit the upregulation of autophagy in the present rodent model. Furthermore, ceramide was accumulated in lung tissues because of LIR; however, hydrogen altered the accumulation status. Significance Molecular hydrogen was found to be therapeutically effective in the LIR-induced ALI model; the mechanisms of action included modulation of antioxidation and autophagy.
Chapter
As research activities progressed, effects of molecular hydrogen (MH) proved to be more extended than previously expected. In parallel with exploratory experiments with animals, human clinical trials have also been performed and yielded positive results in most cases. Here I begin with animal experiments on ischemia-reperfusion injury, peritonitis to sepsis, wound healing, radiation injuries, organ transplantation, metabolic syndrome and lung diseases, and then proceed to describe clinical trials conducted in Japan including recovery from acute erythematous skin disease, cardiac arrest, stroke, brain disorder (Parkinson’s and Alzheimer’s diseases), diabetes, fatigue and aging, rheumatoid arthritis, blood dialysis, cataract surgery, periodontitis, and finally, recent results from other countries regarding radiation injuries and COVID-19 pneumonia. This is a main chapter of the book.
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Patients with sepsis and sepsis-related complications have a high mortality. Endothelial cell dysfunction plays a central role in sepsis pathophysiological process. In sepsis patients, endothelial cell apoptosis is associated with intracellular calcium overload. Multiple functions in the apoptotic process have been found to be regulated by calcium signaling. Our previous work had proved that LPS-induced cell injury was associated with store-operated calcium (SOC) entry mediated by stromal interaction molecule-1 (STIM 1) in Human umbilical vein endothelial cells (HUVEC), but the underlying molecular mechanism has not been adequately defined. Here we report that the LPS-induced cell injury is related to the calcium overload in HUVEC. SOC entry mediated by calcium release-activated calcium modulator (Orai) 1 and transient receptor potential canonical (TRPC) 1 was associated with LPS-induced calcium overload and cell apoptosis. Bruton’s tyrosine kinase (Btk)/Phospholipase C(PLC) γ/inositol 1,4,5-triphosphate receptor (IP3R) played a major role in regulating calcium overload in LPS-induced HUVEC. Knockdown of Btk markedly inhibited the expressions of Orai 1 and its downstream molecule IP3R but not that of TRPC1 in LPS-induced HUVEC. In mice, knockdown of Btk and Orai 1 inhibited LPS-induced calcium overload, pulmonary vascular endothelial cell (VEC) injury and acute lung injury. These findings demonstrated that Btk acts as a regulator of calcium-dependent signaling, especially in the Orai 1-mediated SOC entry of the LPS-induced VEC.
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The small intestine is known to be particularly sensitive to radiation, and the major limiting factor of radiotherapy is the gastrointestinal syndrome that subsequently develops after its administration. The detrimental effects of radiation are mostly mediated via the overproduction of reactive oxygen species (ROS), especially the hydroxyl radical (·OH). Because hydrogen is a selective ·OH scavenger, we hypothesized that hydrogen might exert a protective effect against radiation-induced intestinal damage. Herein, radiation models were built both in mice and in an intestinal crypt epithelial cell (IEC-6) line. In the animal experiment, we demonstrated that hydrogen-rich saline significantly reduced radiation-induced intestinal mucosal damage, improved intestinal function, and increased the survival rate. In addition, radiation-induced oxidative stress damage and systemic inflammatory response were also mitigated by hydrogen treatment. Moreover, hydrogen treatment decreased cell apoptosis and maintained intestinal epithelial cell proliferation in mice. In vitro experiments using the IEC-6 cell line showed that hydrogen-rich medium significantly inhibited ROS formation, maintained cell viability, and inhibited cell apoptosis. Importantly, hydrogen treatment prevented mitochondrial depolarization, cytochrome c release, and activity of caspase-3, caspase-9, and PARP. Moreover, the decreased expression of Bcl-xl and Bcl-2 and the increased expression of Bax protein were also blocked by hydrogen treatment. In conclusion, our study concurrently demonstrated that hydrogen provides an obviously protective effect on radiation-induced intestinal and cell injuries. Our work demonstrated that this protective effect might be due to the blockage of the mitochondrial apoptotic pathway.
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The present work investigated the antioxidative, anti-inflammatory and pulmonary protective effects of enzymatic- and acid- hydrolysed mycelia polysaccharides (En-MPS and Ac-MPS) from Oudemansiella radicata on LPS-induced acute lung injury (ALI) mice. The results demonstrated that both En-MPS and Ac-MPS showed potential pulmonary protective effects by decreasing serum levels of hs-CRP and C3, increasing pulmonary enzyme values of SOD, GSH-Px, CAT and the level of T-AOC; reducing the activity of MPO; and down-regulating the contents of MDA and LPO. In addition, the levels of TNF-ɑ, IL-1β, and IL-6 in BALF of mice treated with En-MPS at a dosage of 400 mg/kg/d were significantly lower than those in the ALI mice. The in vitro antioxidant effects also showed that the En-MPS was more effective than Ac-MPS. Furthermore, the physical properties of polysaccharides were also investigated by GC, HPGPC, FT-IR and NMR. These results indicated that both En-MPS and Ac-MPS possessed potent antioxidant and anti-inflammatory activities, which could be used as an ingestible drug in preventing lung injury.
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Acute respiratory distress syndrome (ARDS) is a devastating disorder that is characterized by increased vascular endothelial permeability and inflammation. Unfortunately, no effective treatment beyond supportive care is available for ARDS. Astilbin, a flavonoid compound isolated from Rhizoma Smilacis Glabrae, has been used for anti-hepatic, anti-arthritic, and anti-renal injury treatments. This study examined the effects of Astilbin on pulmonary inflammatory activation and endothelial cell barrier dysfunction caused by Gram-negative bacterial endotoxin lipopolysaccharide (LPS). Endothelial cells from human umbilical veins or male Kunming mice were pretreated with Astilbin 24h before LPS stimulation. Results showed that Astilbin significantly attenuated the pulmonary histopathological changes and neutrophil infiltration 6h after the LPS challenge. Astilbin suppressed the activities of myeloperoxidase and malondialdehyde, as well as the expression of tumor necrosis factor-α and interleukin-6 in vivo and in vitro. As indices of pulmonary edema, lung wet-to-dry weight ratios, were markedly decreased by Astilbin pretreatment. Western blot analysis also showed that Astilbin inhibited LPS-induced activation of mitogen-activated protein kinase (MAPK) pathways in lung tissues. Furthermore, Astilbin significantly inhibited the activity of heparanase and reduced the production of heparan sulfate in the blood serum as determined by ELISA. These findings indicated that Astilbin can alleviate LPS-induced ARDS, which potentially contributed to the suppression of MAPK pathway activation and the degradation of endothelial glycocalyx.
Article
Objective To reproduce and evaluate a rat model of inhalation lung injury caused by black gunpowder smog. Methods The smog composition was analyzed and a rat model of inhalation lung injury was reproduced. Forty two healthy male Wistar rats were randomly divided into normal control (NC) group and 1h, 2h, 6h, 24h, 48h and 96h after inhalation group (n=6). The arterial blood gas, wet to dry weight ratio (W/D) of lung, leukocyte count, and protein concentration in broncho-alveolar lavage fluid (BALF) were determined. Macroscopic and microscopic changes in lung tissue were observed. Results The composition of black gunpowder smog was composed mainly of CO2 and CO, and their concentrations remained stable within 12 minutes. Smog inhalation caused a significant hypoxemia, the concentration of blood COHb reached a peak value 1h, and the W/D of lung reached peak value 2h after inhalation (P<0.05). The amount of leukocytes and content of protein in BALF increased significantly within 24h after inhalation (P<0.05). Histopathological observation showed diffuse hemorrhage, edema and inflammatory cell infiltration in lung tissue as manifestations of acute lung injury, and the injury did not recover at 96h after inhalation. Conclusion The rat model of inhalation lung injury can be reproduced using black gunpowder smog, and it has the advantages of its readiness for reproduction, reliability and stability, and it could be used for the experiment of inhalation injury in a battlefield environment.
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[Objective] To investigate the protective effects of edaravone treatment on lipopolysaccharide (LPS)-induced acute lung injury (ALI). [Methods] Fifty-four mice were randomized into 3 groups: normal group (Sham operation), control group (LPS + physiological saline) and experiment group (LPS + Edaravone). The mice in control and experiment group were induced ALI by intra-tracheal LPS solution (2 mg/kg body weight). The mice in experiment group were given Edaravone solution (2 mg/mL) 2.5 mL/kg body weight and the mice in control group were given saline at the same volume. At 24 h post-injury, 6 mice in each group were killed and their lung tissues were isolated and assayed for malon-aldehyde (MDA), superoxide dismutase activity (SOD), TNF-α, IL-1β, IL-6 and histological examination. The remained 12 mice in each group were used for monitoring the survival rate of mice every 12 h for 4 days. [Results] Our experiments exhibited that the survival rate of experiment group was higher than control group (P < 0.05). After LPS stimulation, MDA content in the lung tissues was increased from (1.41 ± 0.119) nmol/mg to (4.48 ± 0.159) nmol/mg, whereas it was decreased to (2.56 ± 0.157) nmol/mg in the Edaravone treatment group (P < 0.01). After LPS stimulation, SOD activity in the lung tissues was decreased from (12.86 ± 1.49) U/mg to (8.95 ± 1.02) U/mg, whereas it was increased to (8.95 ± 1.02) nmol/mg in the Edaravone treatment group (P < 0.01). After LPS stimulation, TNF-α, IL-1β and IL-6 content in the lung tissues was increased from (47.89 ± 4.71) pg/mg, (79.39 ± 3.45) pg/mg and (81.9 ± 6.39) pg/mg to (300.48 ± 12.18) pg/mg, (717.99 ± 35.01) pg/mg and (428.99 ± 21.89) pg/mg, whereas they were decreased to (191.84 ± 6.43) pg/mg, (236.87 ± 13.46) pg/mg and (136.92 ± 12.47) pg/mg in the Edaravone treatment group. Edaravone treatment further attenuated lung edema, inflammatory cell infiltrations, widened alveolar septum, and diffuse hemorrhage. [Conclusions] In conclusion, our data demonstrated that ameliorated LPS-induced ALI.
Article
Inflammation response and oxidative stress have been reported to be involved in the pathogenesis of acute lung injury (ALI). Accordingly, anti-inflammatory treatment is proposed to be a possible efficient therapeutic strategy for ALI. The purpose of our present study was to evaluate the anti-inflammatory efficacy of trillin (Tr) on ALI induced by lipopolysaccharide (LPS) in mice and explore the underlying mechanism. BALB/c mice received Tr (50, 100 mg/kg) intraperitoneally 1 h prior to the intratracheal instillation of lipopolysaccharide (LPS) challenge. Pretreatment with Tr at the dose of 50, 100 mg/kg markedly ameliorated lung wet-to-dry weight (W/D) ratio, myeloperoxidase (MPO) activity and pulmonary histopathological conditions. In addition, the protective efficacy of Tr might be attributed to the down-regulations of neutrophil infiltration, malondialdehyde (MDA), inflammatory cytokines and the up-regulations of uper-oxide dismutase (SOD), catalase(CAT), glutathione(GSH), Glutathione Peroxidase(GSH-Px) in bronchoalveolar lavage fluid (BALF). Meanwhile, our study revealed some correlations between (NF-E2-related factor 2) Nrf2/heme oxygenase (HO)-1/nuclear factor-kappa B (NF-κB) pathways and the beneficial effect of Tr, as evidenced by the significant up-regulations of HO-1 and Nrf2 protein expressions as well as the down-regulations of p-NF-κB and p-inhibitor of NF-κB (IκB) in lung tissues. Taken together, our results indicated that Tr exhibited protective effect on LPS-induced ALI by the regulations of related inflammatory events via the activations of Nrf2, HO-1 and NF-κB pathway. The current study indicated that Tr could be a potentially effective candidate medicine for the treatment of ALI.
Article
Puerarin, a kind of traditional Chinese medicine, possesses immunomodulatory property. However, the immunomodulatory effects of puerarin on smoke inhalation injury have not been determined. The aim of the current study was to investigate the therapeutic efficacy of puerarin on gunpowder smog-induced acute lung injury in rats via regulation of Th1/Th2/Th17 expression. Wistar rats were equally randomized to four groups (normal control group, puerarin control group, smoke inhalation injury group, puerarin treatment plus smoke inhalation injury group). The severity of lung injury was evaluated by histopathology, myeloperoxidase (MPO) activity in lung homogenates, cell counting in bronchoalveolar lavage fluid (BALF), and lung vascular permeability parameters including lung wet to dry weight ratio and protein concentration in BALF. Flow cytometry was used to analyze the expression of Th1/Th2/Th17 lymphocytes in blood of rats. Puerarin showed significant therapeutic effects against neutrophil infiltration and tissue injury, as evidenced by histopathological findings and MPO activity. Lung vascular permeability was also relieved by puerarin administration. Additionally, puerarin significantly decreased the number of neutrophils and lymphocytes in BALF compared with smoke inhalation injury group. Furthermore, puerarin increased Th1 immunity and reduced Th2 and Th17 responses and thereby altering the Th1/Th2/Th17 imbalance induced by smoke inhalation. Our findings suggested that puerarin suppressed inflammatory responses in gunpowder smog-induced acute lung injury by regulation of Th1/Th2/Th17 expression, and may be a potential therapeutic agent for smoke inhalation injury. Copyright © 2015 Elsevier B.V. All rights reserved.
Article
Dexmedetomidine (Dex) is widely used for sedation in intensive care units and can be used as an adjunct to anesthetics. Previous studies have demonstrated that Dex has anti-inflammatory property. In this study, we aim to explore the potential therapeutic effects and mechanisms of Dex on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. To induce ALI, mice were intraperitoneally injected with LPS, while Dex was treated 1 h before LPS injection. The inflammation of lung tissues was evaluated by HE stain and bronchoalveolar lavage fluid (BALF) was obtained after 6 h to measure protein concentrations. We also used an enzyme-linked immunosorbent assay to detect the secretion levels of proinflammatory cytokines in the serum. Western blotting method was adopted to observe changes of mitogen-activated protein kinases and downstream nuclear transcription factors. The results showed that pretreatment with Dex considerably reduced neutrophil infiltration and pulmonary edema, and significantly reduced protein concentrations in the BALF, as well as suppressed LPS-induced elevation of proinflammatory cytokines (TNF-α and IL-1β) in the serum. In addition, we observed that the molecular mechanism of Dex-mediated anti-inflammation is associated with decreasing phosphorylation of MKK4, MMK3/6, ERK1/2, p38MAPK and JNK, and diminishing activation of Elk-1, c-Jun and ATF-2. Dex could attenuate ALI induced by LPS in mice and this effect may be mediated through inhibition of MAPK pathway. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
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With the development of thoracic surgeries, one-lung ventilation (OLV) has been routinely used to facilitate surgical exposure. However, OLV can cause lung injury during the surgical process and becomes an important factor affecting the outcomes. To date, effective treatments for the prevention of lung injury caused by OLV are lacking. Hydrogen has been demonstrated to have effective protection against tissue injuries caused by oxidative stress, inflammation, and apoptosis. This study investigated the efficacy of hydrogen water consumption on the prevention of lung injury induced by OLV in rats. Male Sprague-Dawley rats (n = 32, 240-260 g) were divided randomly into the following four groups: sham group, sham + H2 group, OLV group, OLV + H2 group. The rats drank hydrogen water or degassed hydrogen water for 4 wk before the operation and received OLV for 60 min and two-lung ventilation for 60 min. Lung tissues were assayed for wet-to-dry ratio, oxidative stress variables, proinflammatory cytokines, and hematoxylin-eosin staining. Hydrogen water consumption reduced wet-to-dry weight ratio, malondialdehyde and myeloperoxidase activity and decreased the concentration of TNF-α, IL-1β, and IL-6 in the lung tissues compared with sham group and sham + H2 group. Hydrogen water consumption further attenuated NF-κB activation and caused histopathologic alterations. Our data demonstrated that hydrogen water consumption ameliorated OLV-induced lung injury, and it may exert its protective role by its anti-inflammation, antioxidation and reducing NF-κB activity in the lung tissues. Copyright © 2015 Elsevier Inc. All rights reserved.
Article
Objective: To study the early effects of ulinastatin (UTI) by aerosol inhalation on rabbits with acute lung injury induced by LPS, and to observe the early diagnostic value of 320-slice CT. Methods: According to the random number table, 18 specific pathogen free New Zealand white rabbits were divided into normal control group, group LPS, and group UTI, with 6 rabbits in each group. Rabbits in group LPS and group UTI were given 15 mL lipopolysaccharide (0.16 mg/mL, in the dose of 0.8 mg/kg) to reproduce acute lung injury model. Rabbits in normal control group were given equal volume of normal saline. Rabbits in UTI group were treated with UTI by aerosol inhalation for 10 min from 30 min after injury, while those in the other two groups received normal saline by aerosol inhalation. Rabbits in group LPS and group UTI were scanned by 320-slice CT at post injury hour (PIH) 6 and 24. After anesthesia, heart blood of rabbits in group LPS and group UTI was collected for determination of serum levels of TNF-α, IL-1β, and IL-6 by ELISA at PBH 24. At PBH 24, lung tissue samples were harvested for gross observation and histomorphological observation, measurement of wet to dry weight ratio, and detection of mRNA expressions of TNF-α, IL-1β, and IL-6 with RT-PCR. Above-mentioned indexes were detected in rabbits of normal control group at the same time point. Data were processed with one-way analysis of variance and LSD test. Results: (1) CT perfusion (CTP) image. The difference in CTP image of rabbits in group LPS between PBH 6 and PBH 24 was obvious, while that of rabbits in group UTI and normal control group was slight and not obvious respectively. (2) There were statistically significant differences in the serum levels of TNF-α, IL-1β, and IL-6 of rabbits among the three groups (with F values from 843.896 to 2 564.336, P values below 0.001). The serum levels of TNF-α, IL-1β, and IL-6 in group UTI were respectively (225 ± 9), (190 ± 8), (227 ± 6) pg/mL, and they were significantly lower than those in group LPS [(710 ± 25), (306 ± 16), (422 ± 16) pg/mL, with P values below 0.001]. (3) Gross observation. In group UTI, the degrees of pulmonary edema and pneumorrhagia of rabbits were lower than those in group LSP. (4) Histological observation. The damage to alveolar wall in group UTI was milder, and alveolar space hemorrhage and inflammatory cell infiltration were significantly less intense as compared with those in group LPS. (5) Compared with that in normal control group, the wet to dry weight ratio of lung tissue was increased in group LPS (P < 0.001). The wet to dry weight ratio of lung tissue in group UTI was significantly higher than that in normal control group but lower than that in group LPS (P values below 0.001). (6) There were statistically significant differences in mRNA levels of TNF-α, IL-1β, and IL-6 in lung tissue of rabbits among three groups (with F values from 24.700 to 69.538, P values below 0.001). The mRNA levels of TNF-α, IL-1β, and IL-6 in lung tissue of rabbits in group UTI were respectively (31.4 ± 2.7), (21.2 ± 3.3), (13.9 ± 2.4) pg/mL, which were significantly lower than those in group LPS [ (58.5 ± 10.0) , (35.1 ± 5.1), (20.7 ± 3.2) pg/mL, P values below 0.001]. Conclusions: UTI by aerosol inhalation can mitigate pulmonary edema and hemorrhage and inhibit inflammatory response. 320-slice CT may be used for detection of early lung injury.
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Polymer materials are well-used for Modern building constructions. Combustion gas of construction material has different components between natural and polymer. Recently research shows, Apart from combustion gas of natural construction material contain CO. species. Also polymer combustion gas include HF, HCN, HBr, HCl, NO, , and additionally. It analyzed that species product from Polymer compounds. These species can act as major disturbance factors in the evacuation from the building's fire. Because, polymer will increase the proportion in building materials. In this research, effects from HF gas are analyzed by pathological method. Histopathologic examination, DNA toxicity evaluation experiments were conducted for mouse exposed by HF standard gas. Exposing HF gas were caused extensive hemorrhage and necrosis in the liver and lung from experimental animals. and blood and DNA damage were analyzed quantitatively by comet assay.
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We and others previously reported that the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 significantly accumulate with age in mouse lung, this is accompanied by elevated phosphorylation of p38. Here, we further investigate whether aging affects activation of p38 signalling and the inflammatory reaction after exposure to lipopolysaccharide (LPS) in lungs of mice in vivo and humans ex vivo. The data showed that activation of p38 peaked at 0.5h and then rapidly declined in young (2-month-old) mouse lung, after intranasal inhalation challenge with LPS. In contract, activation of p38 peaked at 24h and was sustained longer in aged (20-month-old) mice. As well as altered p38, activations of its upstream activator MKK and downstream substrate NF-κB were also changed in lungs of aged mice, which corresponded with the absent in the early phase but delayed increases in concentrations of TNF-α, IL-1β and IL-6. Consistent with the above observations in mice, similar patterns of p38 signalling also occurred in human lungs. Compared with younger lungs from adult-middle aged subjects, the activation of p38, MKK and NF-κB, as well as the production of pro-inflammatory cytokines were significantly increased in lungs of older subjects ex vivo. Exposure of human lung cells to LPS induced rapid activation of p38, MKK and NF-κB in these cells from adultmiddle aged subjects, but not older subjects, with increases in production of the pro-inflammatory cytokines. The LPS-induced rapid activation in lung cells from adult-middle aged subjects occurred as early as 0.25h after exposure, and then declined. Compared with adult-middle aged subjects, LPS exposure did not induce marked changes in the early phase, either in the activation of p38, MKK and NF-κB, or in the production of TNF-α, IL-1β or IL-6 in lung cells from older subjects. In contrast, these changes occurred relatively late, peaked at 16h and were sustained longer in lungs of older subjects. These data support the hypothesis that the sustained activation of the p38 signalling pathway at baseline and the absence in the early phase but delayed of p38 signalling pathway response to LPS in the elderly may play important roles in increased susceptibility of aged lungs to inflammatory injury.
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Mechanical ventilation (MV) can provoke oxidative stress and an inflammatory response, and subsequently cause ventilator-induced lung injury (VILI), a major cause of mortality and morbidity of patients in the intensive care unit. Inhaled hydrogen can act as an antioxidant and may be useful as a novel therapeutic gas. We hypothesized that, owing to its antioxidant and anti-inflammatory properties, inhaled hydrogen therapy could ameliorate VILI. VILI was generated in male C57BL6 mice by performing a tracheostomy and placing the mice on a mechanical ventilator (tidal volume of 30 ml/kg without positive end-expiratory pressure, FiO(2) 0.21). The mice were randomly assigned to treatment groups and subjected to VILI with delivery of either 2% nitrogen or 2% hydrogen in air. Sham animals were given same gas treatments for two hours (n = 8 for each group). The effects of VILI induced by less invasive and longer exposure to MV (tidal volume of 10 ml/kg, 5 hours, FiO(2) 0.21) were also investigated (n = 6 for each group). Lung injury score, wet/dry ratio, arterial oxygen tension, oxidative injury, and expression of pro-inflammatory mediators and apoptotic genes were assessed at the endpoint of two hours using the high-tidal volume protocol. Gas exchange and apoptosis were assessed at the endpoint of five hours using the low-tidal volume protocol. Ventilation (30 ml/kg) with 2% nitrogen in air for 2 hours resulted in deterioration of lung function, increased lung edema, and infiltration of inflammatory cells. In contrast, ventilation with 2% hydrogen in air significantly ameliorated these acute lung injuries. Hydrogen treatment significantly inhibited upregulation of the mRNAs for pro-inflammatory mediators and induced antiapoptotic genes. In the lungs treated with hydrogen, there was less malondialdehyde compared with lungs treated with nitrogen. Similarly, longer exposure to mechanical ventilation within lower tidal volume (10 mg/kg, five hours) caused lung injury including bronchial epithelial apoptosis. Hydrogen improved gas exchange and reduced VILI-induced apoptosis. Inhaled hydrogen gas effectively reduced VILI-associated inflammatory responses, at both a local and systemic level, via its antioxidant, anti-inflammatory and antiapoptotic effects.
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This study has evaluated the ability of the semiessential amino acid taurine to attenuate lipopolysaccharide (LPS)-induced lung inflammation, oxidative stress and apoptosis in a small animal model. For this purpose, bacterial LPS (0.02mg in phosphate buffered saline (PBS) pH 7.4) was instilled intratracheally into female Golden Syrian hamsters, before or after a 3-day intraperitoneal treatment with a single dose (50mg/kg in PBS pH 7.4) of taurine. At 24h after the last treatment, lung tissue and bronchoalveolar lavage fluid (BALF) samples were collected. In comparison to samples from animals receiving only PBS pH 7.4, serving as controls, those of LPS-stimulated animals exhibited a higher count of both total leukocytes and neutrophils in the BALF, and increased incidence of apoptosis, depletion of intracellular glutathione and evidence of inflammation confined to the parenchyma in the lung. In addition, LPS caused cells in the BALF to exhibit a higher expression of tumor necrosis factor-1, a higher activity of caspase-3, marked lipid peroxidation, and altered activities of catalase, glutathione peroxidase and superoxide dismutase relative to control samples. In contrast, a treatment with taurine was found to significantly attenuate all of the cellular and biochemical alterations induced by LPS, more so when given before rather than after the endotoxin. The present results suggest that taurine possesses intrinsic antiinflammatory and antioxidant properties that may be of benefit against the deleterious actions of LPS in the lung.
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This study is to examine if hydrogen-rich saline reduced amyloid-beta (A beta) induced neural inflammation and oxidative stress in a rat model by attenuation of activation of JNK and NF-kappa B. Sprague-Dawley male rats (n = 18,280-330 g) were divided into three groups, sham operated, A beta 1-42 injected and A beta 1-42 plus hydrogen-rich saline treated animals. Hydrogen-rich saline (5 ml/kg, i.p., daily) was injected for 10 days after intraventricular injection of A beta 1-42. The levels of IL-1 beta were assessed by ELISA analysis, 8-OH-dG by immunohistochemistry in the brain slides, and JNK and NF-kappa B by immunohistochemistry and western blotting. After A beta 1-42 injection, the level of IL-1 beta, 8-OH-dG, JNK and NF-kappa B all increased in brain tissues, while hydrogen-rich saline treatment decreased the level of IL-1 beta, 8-OH-dG and the activation of INK and NF-kappa B. In conclusion, hydrogen-rich saline prevented A beta-induced neuroinflammation and oxidative stress, possibly by attenuatation of activation of c-Jun NH(2)-terminal kinase (JNK) and nuclear factor-kappa B (NF-kappa B) in this rat model. (C) 2011 Elsevier Ireland Ltd. All rights reserved.
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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.
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Hypoxia-ischemia (HI) brain injury is a major cause of neuronal cell death especially apoptosis in the perinatal period. This study was designated to examine the effect of hydrogen therapy on apoptosis in an established neonatal HI rat pup model. Seven-day-old rat pups were subjected to left common carotid artery ligation and then 90 min hypoxia (8% oxygen at 37 C). Immediately after HI insult, pups were placed into a chamber filled with 2% H(2) for 30 min, 60 min, or 120 min, respectively. 24 h after 2% H2 therapy, the pups were decapitated and brain injury was assessed by 2,3,5-triphenyltetrazoliumchloride (TTC), Nissl, and TUNEL staining, as well as caspase-3, caspase-12 activities in the cortex and hippocampus. H(2) treatment in a duration-dependent manner significantly reduced the number of positive TUNEL cells and suppressed caspase-3 and -12 activities. These results indicated H(2) administration after HI appeared to provide brain protection via inhibition of neuronal apoptosis. (C) 2008 Elsevier Ireland Ltd. All rights reserved.
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The present study aimed to investigate the effect of the new tyrosine kinase inhibitor, nilotinib on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats and explore its possible mechanisms. Male Sprague-Dawley rats were given nilotinib (10mg/kg) by oral gavage twice daily for 1week prior to exposure to aerosolized LPS. At 24h after LPS exposure, bronchoalveolar lavage fluid (BALF) samples and lung tissue were collected. The lung wet/dry weight (W/D) ratio, protein level and the number of inflammatory cells in the BALF were determined. Optical microscopy was performed to examine the pathological changes in lungs. Malondialdehyde (MDA) content, superoxidase dismutase (SOD) and reduced glutathione (GSH) activities as well as nitrite/nitrate (NO(2)(-)/NO(3)(-)) levels were measured in lung tissues. The expression of inflammatory cytokines, tumor necrosis factor-α (TNF-α), transforming growth factor-β(1) (TGF-β(1)) and inducible nitric oxide synthase (iNOS) were determined in lung tissues. Treatment with nilotinib prior to LPS exposure significantly attenuated the LPS-induced pulmonary edema, as it significantly decreased lung W/D ratio, protein concentration and the accumulation of the inflammatory cells in the BALF. This was supported by the histopathological examination which revealed marked attenuation of LPS-induced ALI in nilotinib treated rats. In addition, nilotinib significantly increased SOD and GSH activities with significant decrease in MDA content in the lung. Nilotinib also reduced LPS mediated overproduction of pulmonary NO(2)(-)/NO(3)(-) levels. Importantly, nilotinib caused down-regulation of the inflammatory cytokines TNF-α, TGF-β(1) and iNOS levels in the lung. Taken together, these results demonstrate the protective effects of nilotinib against the LPS-induced ALI. This effect can be attributed to nilotinib ability to counteract the inflammatory cells infiltration and hence ROS generation and regulate cytokine effects.
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Acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS) are common syndromes that affect both clinical and surgical patients. This study describes the effects of a potent and specific N-methyl-d-aspartate receptor antagonist (MK-801) against oxidative stress in acute lung injury induced by intratracheal lipopolysaccharide (LPS) injection. This study was performed using male Wistar rats weighing 200-250g. Rats were randomly divided into four groups: control with isotonic saline instillation (n=6); LPS (100μg/100g of body weight) treated with saline (n=6); LPS treated with MK-801 (0.3mg/kg, intraperitoneally; n=6); LPS treated with MK-801 (0.3mg/kg, intratracheally; n=6). Twelve hours after the LPS instillation, rats were anesthetized and a bronchoalveolar lavage (BAL) was performed in order to determine the alveolar-capillary membrane alterations and the inflammatory infiltrate level. Blood and lung samples were isolated and assayed for oxidative stress variables and histopathologic analysis. The use of MK-801 decreased bronchoalveolar lavage fluid protein, LDH activity and inflammatory cells. Indeed, the treatment with MK-801 significantly attenuated lung oxidative damage and histopathologic alterations after LPS instillation. Our data provide the first experimental demonstration that MK-801 decreases oxidative stress and limits inflammatory response and alveolar disarray in lipopolysaccharide-induced acute lung injury.