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Mulberrin reduces liver fibrosis in CCl 4 -treated mice. (A) H&E, Sirius red and Masson's Trichrome staining of liver sections. (B) Quantification for fibrosis score. (C,D) Quantification for fibrotic areas by Sirius red and Masson's Trichrome staining, respectively. Serum contents of (E) hyaluronic acid, (F) procollagen III (PC III) (G) and Collagen IV were measured. Representative data were expressed as mean ± SEM (n = 5 for histological analysis, or 10 for biological analysis per group). Magnification: × 100. **P < 0.01 and ***P < 0.001 vs the Oil/Veh group; +P < 0.05, ++P < 0.01 and +++P < 0.001 vs the CCl 4 /Veh group. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Source publication
Mulberrin (Mul) is a key component of the traditional Chinese medicine Romulus Mori with various biological functions. However, the effects of Mul on liver fibrosis have not been addressed, and thus were investigated in our present study, as well as the underlying mechanisms. Here, we found that Mul administration significantly ameliorated carbon t...
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... hepatic injury. Sirius red and Masson's Trichrome staining demonstrated that CCl 4 -challenged mice exhibited severer collagen deposition and fibrosis in liver sections than that of the Oil/Veh group. Notably, these histological alterations and hepatic fibrosis were significantly mitigated by Mul treatments via a concentration-dependent fashion ( Fig. 2A-D). Furthermore, the levels of three key liver fibrosis hallmarks including hyaluronic acid, PC III and Collagen IV were strongly promoted by CCl 4 , while being strongly ameliorated in Mul-treated mice compared with the model group ( mice compared with the model group ( Fig. 3F). Therefore, Mul could restrain TGF-β1/SMADs signaling to ...
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... shown in Fig. 7E, TGF-β1 stimulation led to inflammatory response in LX-2 cells, proved by the markedly enhanced gene expression levels of TNF-α, IL-1β, IL-6, IL-18, MCP-1 and CXCL10; however, such event caused by TGF-β1 was significantly abrogated following Mul exposure. Such anti-inflammatory effects mediated by Mul were validated in mouse primary HSCs upon TGF-β1 stimulation (Supplementary Fig. 2E). Moreover, HO-1 and NQO1 gene expression levels were highly decreased in LX-2 cells and mouse primary HSCs stimulated by TGF-β1, while being efficiently reversed upon Mul co-treatment. ...
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... HSCs upon TGF-β1 stimulation (Supplementary Fig. 2E). Moreover, HO-1 and NQO1 gene expression levels were highly decreased in LX-2 cells and mouse primary HSCs stimulated by TGF-β1, while being efficiently reversed upon Mul co-treatment. In contrast, NOX2 and NOX4 expression levels were greatly abolished by Mul in a dose-dependent fashion ( Fig. 7F and Supplementary Fig. 2F). In response to TGF-β1, nuclear Nrf2 protein expression levels were significantly weakened, whereas being markedly rescued after Mul exposure. Opposite profile was detected in the expression of cytoplastic Nrf2 both in LX-2 and primary mouse HSCs ( Fig. 7G and Supplementary Fig. 2G). These in vitro findings suggested that Mul could ...
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... were greatly abolished by Mul in a dose-dependent fashion ( Fig. 7F and Supplementary Fig. 2F). In response to TGF-β1, nuclear Nrf2 protein expression levels were significantly weakened, whereas being markedly rescued after Mul exposure. Opposite profile was detected in the expression of cytoplastic Nrf2 both in LX-2 and primary mouse HSCs ( Fig. 7G and Supplementary Fig. 2G). These in vitro findings suggested that Mul could restrain the activation, inflammation and oxidative stress in ...
Citations
... which in turn inhibits glucose production, attenuates hepatic fibrosis by reducing CTGFmediated deposition of collagen fibers, and attenuates the accumulation of pro-inflammatory factors, cytokines, and chemokines mediated by c-jun phosphorylation, thereby reducing the inflammatory response (26). Another research team discovered a natural compound 'mulberrin' in mulberry branches that can effectively target activation of TRIM31, reduce oxidative stress and liver inflammation, thereby alleviating NASH (27). However, instead of using the traditional RING structural domain, TRIM16 uses the B-box structural domain to perform its ubiquitination function. ...
Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases and is closely associated with metabolic abnormalities. The causes of NAFLD are exceedingly complicated, and it is known that a variety of signaling pathways, endoplasmic reticulum stress, and mitochondrial dysfunction play a role in the pathogenesis of NAFLD. Recent studies have shown that ubiquitination and deubiquitination are involved in the regulation of the NAFLD pathophysiology. Protein ubiquitination is a dynamic and diverse post-translational alteration that affects various cellular biological processes. Numerous disorders, including NAFLD, exhibit imbalances in ubiquitination and deubiquitination. To highlight the significance of this post-translational modification in the pathogenesis of NAFLD and to aid in the development of new therapeutic approaches for the disease, we will discuss the role of enzymes involved in the processes of ubiquitination and deubiquitination, specifically E3 ubiquitin ligases and deubiquitinating enzymes that are important in the regulation of NAFLD.
... Inhibiting HSC activation serves as a potential therapeutic approach for treating liver fibrosis. The intricate and diverse chemical structures of natural products have demonstrated significant potential for combating liver fibrosis, and these products are promising candidates for novel drug development [26][27][28][29][30]. ...
Background
Liver fibrosis is a representative scarring response that can ultimately lead to liver cancer. However, relevant antifibrotic drugs for the effective treatment of liver fibrosis in humans have not yet been identified. Chikusetsusaponin IVa (CS-IVa) is derived from natural products and exhibits multiple biological activities; however, its efficacy and potential mechanism of action against liver fibrosis remains unclear.
Purpose
This study aimed to examine the antifibrotic properties and potential mechanisms of action of CS-IVa.
Methods
We constructed two mature mouse models (CCl 4 challenge and bile duct ligation) to evaluate the antifibrotic properties of CS-IVa in vivo. Proteomics analysis and transforming growth factor β1 (TGF-β1)-activated LX-2 cells were used to elucidate the potential effects and mechanisms. Molecular docking, surface plasmon resonance (SPR), and cellular thermal shift assay (CETSA) were used to detect the affinity and binding between CS-IVa and its target.
Results
We found that CS-IVa significantly alleviated liver fibrosis and injury by downregulating yes-associated protein (YAP) and tafazzin (TAZ) expression. In an in vitro model, CS-IVa suppressed TGF-β1-induced hepatic stellate cell (HSC) activation, as well as the mRNA and protein expression of COL1A1, α-SMA, YAP, and TAZ. Moreover, specific knockdown or inhibition of YAP did not enhance the suppressive effect of CS-IVa on HSC activation or fibrosis-associated protein expression. Molecular docking, SPR, and CETSA showed that CS-IVa could directly bind to YAP.
Conclusion
These findings demonstrated that the administration of CS-IVa effectively alleviated liver fibrosis by suppressing the YAP/TAZ pathways. In addition, CS-IVa could directly bind to YAP and act as a YAP inhibitor.
... Ecological imbalances and disorders of intestinal microflora can lead to damage to the intestinal barrier and intestinal inflammation, which in turn can lead to intestinal mucosal damage (25). Chinese medicine can improve chronic liver disease through intestinal flora (26)(27)(28). We found that the traditional Chinese medicine FAE had a substantial influence on the diversity of intestinal flora in mice, and the number of OTUs in the FAE group differed greatly from that in the other two groups. ...
Objectives
To explore the mechanism underlying the effect of Fructus Akebiae (FAE) against hepatic fibrosis in mice through combined network pharmacology, liver metabolomics, and 16S rDNA analyses of the gut microbiota.
Methods
In this study, we randomly divided mice into the control, model, FAE high-dose, FAE medium-dose, and FAE low-dose groups to analyze the pathological changes in the hepatic fibrosis and levels of the α-SMA, collagen 1, Nuclear Factor Kappa B (NF-κ B), Toll Like Receptor 4 (TLR4). The gut microbiota was analyzed through 16S rDNA sequencing analysis of liver metabolites using liquid chromatography-mass spectrometry. Furthermore, network pharmacology was used to determine the specific molecular regulation mechanism of FAE in hepatic fibrosis treatment.
Results
FAE treatment markedly improved the pathological changes in the hepatic fibrosis. Analysis revealed that FAE administration reversed the carbon tetrachloride (CCl4)-induced dysbiosis by increasing the abundance of Akkermansia and reducing that of Cyanobacteria. Additionally, metabolomic analysis showed that FAE treatment reversed the CCl4-induced metabolic disorders by regulating amino and nucleotide sugar metabolism. Furthermore, correlation analysis showed that Akkermansia and Verrucomicobiota were closely related to D-tolasaccharide and maltotetraose saccharide. Moreover, network pharmacology indicated that FAE might regulate the signaling pathway through the JUN/CASP3/NOS3/PTGS2/HSP90AA1 during treatment.
Conclusion
FAE may be a promising treatment for hepatic fibrosis, and its protective effects are associated with improvements in the microbiome and metabolic disorders.
... Numerous studies have highlighted the critical role of Trim31 in the regulation of fibrosis. The overexpression of Trim31 has been shown to ameliorate the fibrotic process in the kidney [11,14] and liver [45]. However, it is still unclear whether Trim31 plays a role in the regulation of fibrosis in the heart. ...
Tripartite motif-containing protein 31 (Trim31) is known to be involved in various pathological conditions, including heart diseases. Nonetheless, its specific involvement in heart failure (HF) has yet to be determined. In this study, we examined the function and mechanism of Trim31 in HF by using mice with cardiac-specific knockout (cKO) of Trim31. The HF mouse model was induced via the subcutaneous injection of isoproterenol (ISO). We observed a decrease in Trim31 expression in the heart tissues of mice with HF. Compared with wild-type (WT) mice, Trim31 cKO mice presented more severe characteristics of HF, including worsened cardiac dysfunction, hypertrophy, and fibrosis. However, these symptoms in Trim31 cKO mice were significantly reversed when they received an intramyocardial injection of recombinant adeno-associated virus (AAV) expressing Trim31. Excessive activation of the NLRP3 inflammasome, manifested by increased levels of NLRP3, ASC, cleaved Caspase-1, cleaved GSDMD, IL-1β, and IL-18, was observed in Trim31 cKO mice with HF. However, Trim31 overexpression effectively reversed the NLRP3 inflammasome activation in Trim31 cKO mice with HF. Selective inhibition of the NLRP3 inflammasome with the NLRP3 inhibitor MCC950 effectively reversed the worsened cardiac dysfunction, hypertrophy, and fibrosis observed in Trim31 cKO mice with HF. Overall, the findings from this study reveal a crucial role of Trim31 in HF. Trim31 deficiency may contribute to the progression of HF by promoting cardiac hypertrophy, fibrosis, and inflammation by facilitating the activation of the NLRP3 inflammasome. Therefore, Trim31 may hold significant potential as a therapeutic target for the treatment of HF.
... Interestingly, the uncontrolled secretion of inflammatory factors and chemokines by macrophages are important driving force for HF formation 10 . In addition, inflammation-induced changes in the liver microenvironment lead to dysregulation of liver function, reducing the capacity for self-repair and accelerating HF progression 11 . ...
Kupffer cells (KCs), as residents and sentinels of the liver, are involved in the formation of hepatic fibrosis (HF). However, the biological functions of circular RNAs (circRNAs) in KCs to HF have not been determined. In this study, the expression levels of circRNAs, microRNAs, and messenger RNAs (mRNAs) in KCs from a mouse model of HF mice were investigated using microarray and circRNA-Seq analyses. circDcbld2 was identified as a candidate circRNA in HF, as evidenced by its up-regulation in KCs. Silver staining and mass spectrometry showed that Wtap and Igf2bp2 bind to cirDcbld2. The suppression of circDcbld2 expression decreased the KC inflammatory response and oxidative stress and inhibited hepatic stellate cell (HSCs) activation, attenuating mouse liver fibrogenesis. Mechanistically, Wtap mediated the N⁶-methyladenosine (m6A) methylation of circDcbld2, and Igf2bp2 recognized m6A-modified circDcbld2 and increased its stability. circDcbld2 contributes to the occurrence of HF by binding miR-144-3p/Et-1 to regulate the inflammatory response and oxidative stress. These findings indicate that circDcbld2 functions via the m6A/circDcbld2/miR-144-3p/Et-1 axis and may act as a potential biomarker for HF treatment.
... Two more recent studies have demonstrated that TRIM31 alleviates NAFLD and NASH pathologies by targeted degradation of rhomboid 5 homolog 2 (Rhbdf2) (35) and TAK1 (36) in the liver. TRIM31 is also responsible for the antifibrotic effects of mulberrin (a bioactive phytochemical from the traditional Chinese medicine Ramulus Mori) in CCl 4 -induced liver fibrosis (37). These lines of evidence illustrate the therapeutic potential of targeting TRIM family members in different stages of NAFLD. ...
Nonalcoholic fatty liver disease (NAFLD) encompasses a disease continuum from simple steatosis to nonalcoholic
steatohepatitis (NASH). However, there are currently no approved pharmacotherapies for NAFLD, although several drugs
are in advanced stages of clinical development. Because of the complex pathophysiology and heterogeneity of NAFLD,
the identification of potential therapeutic targets is clinically important. Here, we demonstrated that tripartite motif
56 (TRIM56) protein abundance was markedly downregulated in the livers of individuals with NAFLD and of mice fed
a high-fat diet. Hepatocyte-specific ablation of TRIM56 exacerbated the progression of NAFLD, while hepatic TRIM56
overexpression suppressed it. Integrative analyses of interactome and transcriptome profiling revealed a pivotal role of
TRIM56 in lipid metabolism and identified the lipogenesis factor fatty acid synthase (FASN) as a direct binding partner
of TRIM56. TRIM56 directly interacted with FASN and triggered its K48-linked ubiquitination–dependent degradation.
Finally, using artificial intelligence–based virtual screening, we discovered an orally bioavailable small-molecule inhibitor
of FASN (named FASstatin) that potentiates TRIM56-mediated FASN ubiquitination. Therapeutic administration of
FASstatin improved NAFLD and NASH pathologies in mice with an optimal safety, tolerability, and pharmacokinetics
profile. Our findings provide proof of concept that targeting the TRIM56/FASN axis in hepatocytes may offer potential
therapeutic avenues to treat NAFLD.
... Furthermore, the MAFLD therapeutic effects of SZ-A were believed to be associated with the activation of several substances as mentioned above. Moreover, relevant studies also suggested that SZ-A could activate the NRF2 axis to inhibit the excitation of hepatic stellate cells, thereby alleviating liver fibrosis [21,22]. It was also observed that PGC1α could regulate the expression of nuclear/mitochondrial genes associated with oxidative phosphorylation by coactivating NRF2, indicating that NRF2 may also act as an important pathway through which SZ-A exerts MAFLD-remedying effects [23]. ...
Background/Objectives: Metabolic-associated fatty liver disease (MAFLD) is one of the most common liver disorders associated with obesity and metabolic syndrome, and poses a significant global health burden with limited effective treatments. The aim of this study was to assess the protective effects of mulberry twig alkaloids (SZ-A) on MAFLD and to further investigate the underlying mechanisms including the specific targets or pathways. Methods: Diet-induced obesity (DIO) and normal mouse models were established by feeding C57Bl/6J mice with a high-fat diet (HFD) or common diet for 12 weeks. SZ-A, dapagliflozin, and placebo were administered to corresponding mouse groups for 8 weeks. Data of fasting blood glucose, glucose tolerance, insulin tolerance, and the body weight of mice were collected at the baseline and termination of the experiment. Serum liver enzymes and lipids were measured by ELISA. Western blotting, qPCR, and pathological section staining were implemented to evaluate the degrees of liver steatosis, fibrosis, and oxidative stress in mice. Results: In DIO mouse models, high-dose SZ-A (800 mg/kg/d) treatment significantly inhibited HFD-induced weight gain, improved insulin tolerance, and reduced serum alanine aminotransferase, total cholesterol, and triglyceride levels compared with placebo. In DIO mice, SZ-A could alleviate the pathological changes of hepatic steatosis and fibrosis compared with placebo. Lipid catabolism and antioxidant stress-related proteins were significantly increased in the livers of the high-dose SZ-A group (p < 0.05). Inhibition of PGC1α could inhibit the function of SZ-A to enhance lipid metabolism in hepatocytes. PGC1α might interact with NRF2 to exert MAFLD-remedying effects. Conclusions: By regulating the expression of PGC1α and its interacting KEAP1/NRF2 pathway in mouse liver cells, SZ-A played important roles in regulating lipid metabolism, inhibiting oxidative stress, and postponing liver fibrosis in mice with MAFLD.
... For IF analysis, the HET-1A cells after treatments were washed with PBS and were then blocked in 10% goat serum (#C0265, Beyotime Biotechnology) containing 0.3% Triton X-100 (#ST797, Beyotime Biotechnology) for 1 h at room temperature and incubated overnight with primary antibody NLRP3 (1:1000 #C1006, Beyotime Biotechnology). Photographs were captured using a fluorescence microscope [31]. ...
Background
Reflux esophagitis (RE) is a disease in which inflammation of the esophageal mucosa owing to the reflux of gastric contents into the esophagus results in cytokine damage. Britannilactone 1-O-acetate (Brt) has anti-inflammatory effects, significantly inhibiting the activation of the NLRP3 inflammasome, leading to a decrease in inflammatory factors including IL-1 β, IL-6, and TNF-α. However, the mechanism underlying its protective effect against RE-induced esophageal injury remains unclear. In the present study, we investigated the protective mechanism of TRIM31 against NLRP3 ubiquitination-induced RE both in vivo and in vitro.
Methods
A model of RE was established in vivo in rats by the method of “4.2 mm pyloric clamp + 2/3 fundoplication”. In vitro, the mod was constructed by using HET-1A (esophageal epithelial cells) and exposing the cells to acid, bile salts, and acidic bile salts. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay was used to screen the concentration of administered drugs, and the viability of HET-1A cells in each group. HE staining was used to assess the degree of pathological damage in esophageal tissues. Toluidine blue staining was used to detect whether the protective function of the esophageal epithelial barrier was damaged and restored. The enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of IL-1 β, IL-6, and TNF-α factors in serum. Immunohistochemistry (IHC) was used to detect the expression level of NLRP3 in esophageal tissues. The molecular docking and Co-immunoprecipitation assay (Co-IP assay) were used to detect the TRIM31 interacts with NLRP3. Western blotting detected the Claudin-4, Claudin-5, The G-protein-coupled receptor calcium-sensitive receptor (CaSR), NLRP3, TRIM31, ASC, C-Caspase1, and Caspase1 protein expression levels.
Results
Brt could alleviate RE inflammatory responses by modulating serum levels of IL-1 β, IL-6, and TNF-α. It also activated the expression of NLRP3, ASC, Caspase 1, and C-Caspase-1 in HET-1A cells. Brt also attenuated TRIM31/NLRP3-induced pathological injury in rats with RE through a molecular mechanism consistent with the in vitro results.
Conclusions
Brt promotes the ubiquitination of NLRP3 through TRIM31 and attenuates esophageal epithelial damage induced by RE caused by acidic bile salt exposure. This study provides valuable insights into the mechanism of action of Brt in the treatment of RE and highlights its promising application in the prevention of NLRP3 inflammatory vesicle-associated inflammatory pathological injury.
Graphical Abstract
Supplementary Information
The online version contains supplementary material available at 10.1186/s13020-024-00986-y.
... The culture medium used was Dulbecco's Modified Eagle Medium supplemented with 10 % FBS and 1 % penicillin/streptomycin. Prior to intervention, LX2 was cultured in vitro and pre-treated 1,25(OH) 2 D 3 (100 nM) for 16 h [21,30]. Subsequently, TGF-β1 (10 ng/mL) was administered to induce LX2 activation for 12 h [31], followed by treatment with VP (10 nM) or siRNA in combination with 1,25 (OH) 2 D 3 (100 nM) for 12 h. Upon completion the experiment, protein and RNA were extracted from the cells and analyzed individually. ...
... Multiple cytokines induced the activation of NF-κB pathway, thus contributed to the enhance of inflammation response, EMT and tumorigenesis. Ge et al. found that the function of Mulberrin against hepatic fibrosis and oxidative stress was depended on the expression of TRIM31 and the suppression of NF-κB pathway and NOD-like receptor protein 3 (NLRP3) inflammasome [49]. In pancreatic cancer and colorectal cancer, TRIM31 played a role in the activation of the NF-κB and the downstream genes [41,50]. ...
Most TRIM family members characterized by the E3-ubiquitin ligases, participate in ubiquitination and tumorigenesis. While there is a dearth of a comprehensive investigation for the entire family in gastric cancer (GC). By combining the TCGA and GEO databases, common TRIM family members (TRIMs) were obtained to investigate gene expression, gene mutations, and clinical prognosis. On the basis of TRIMs, a consensus clustering analysis was conducted, and a risk assessment system and prognostic model were developed. Particularly, TRIM31 with clinical prognostic and diagnostic value was chosen for single-gene bioinformatics analysis, in vitro experimental validation, and immunohistochemical analysis of clinical tissue microarrays. The combined dataset consisted of 66 TRIMs, of which 52 were differentially expressed and 43 were differentially prognostic. Significant survival differences existed between the gene clusters obtained by consensus clustering analysis. Using 4 differentially expressed genes identified by multivariate Cox regression and LASSO regression, a risk scoring system was developed. Higher risk scores were associated with a poorer prognosis, suppressive immune cell infiltration, and drug resistance. Transcriptomic data and clinical sample tissue microarrays confirmed that TRIM31 was highly expressed in GC and associated with a poor prognosis. Pathway enrichment analysis, cell migration and colony formation assay, EdU assay, reactive oxygen species (ROS) assay, and mitochondrial membrane potential assay revealed that TRIM31 may be implicated in cell cycle regulation and oxidative stress-related pathways, contribute to gastric carcinogenesis. This study investigated the whole functional and expression profile and a risk score system based on the TRIM family in GC. Further investigation centered around TRIM31 offers insight into the underlying mechanisms of action exhibited by other members of its family in the context of GC.
Supplementary Information
The online version contains supplementary material available at 10.1186/s40246-024-00631-7.
