Pharmacological preconditioning of mesenchymal stem cells (MSCs) is a promising strategy to enhance their therapeutic efficacy for end-stage liver disease; however, maximizing this benefit remains a major clinical challenge. Senkyunolide H (SNH), a small-molecule compound derived from Angelica sinensis, exhibits anti-inflammatory, antioxidant, and anti-apoptotic properties. Nevertheless, its capacity to optimize MSCs-based therapy for liver disease has not been fully elucidated. Here, we demonstrate that SNH preconditioning significantly enhances the therapeutic efficacy of bone marrow mesenchymal stem cells (BMSCs) in a murine model of liver cirrhosis. Specifically, SNH-pretreated BMSCs markedly alleviated hepatocellular injury, promoted hepatocyte proliferation, and attenuated collagen deposition. Mechanistically, SNH augments the therapeutic potency of BMSCs by partly binding to macrophage erythroblast attacher (MAEA), a subunit of the E3 ubiquitin ligase complex. This interaction stabilizes MAEA, which in turn facilitates the ubiquitination and proteasomal degradation of dual specificity phosphatase 6 (DUSP6), thereby activating ERK/STAT3 signaling and upregulating the secretion of hepatocyte growth factor (HGF). Collectively, our findings highlight SNH preconditioning as a robust approach to enhance the paracrine function and therapeutic potential of BMSCs, and identify MAEA as a novel therapeutic target for BMSCs-based interventions in liver cirrhosis.

ETHNOPHARMACOLOGICAL RELEVANCE:Fuzheng Huayu formula (FZHY) is clinically used for liver fibrosis treatment, but its pharmacological mechanisms remain incompletely understood. AIM OF THE STUDY:This study explored the anti-fibrotic effect of FZHY, particularly its efficacy against cholestatic liver fibrosis, and the underlying molecular mechanisms. MATERIALS AND METHODS:The therapeutic potential of FZHY was evaluated using Mdr2-/- mice and CCl4/2-AAF-induced rats of liver fibrosis model rodents, and GANT61, a Gli1 inhibitor was used as a positive control drug. Histological analysis and double-immunofluorescence techniques were employed to detect the effects of FZHY on ductular reaction and liver fibrosis. In vitro, WB-F344 cells, a hepatic progenitor cell (HPC) line, were transduced with Gli1-overexpressing lentiviral vectors and induced with sodium butyrate (SB) to verify the mechanisms of FZHY. RESULTS:In both Mdr2-/- mice and CCl4/2-AAF-induced rats, FZHY obviously ameliorated liver fibrosis, evidenced by reduced collagen deposition, decreased hepatic hydroxyproline content, and less inflammatory cell infiltration. FZHY reduced the expression of Epcam, CK19, and CK7. Double-immunofluorescence revealed an increased number of CK19+ and OV6+ cells in the CCl4/2-AAF rats, which decreased after FZHY treatment. Mechanistically, FZHY downregulated Gli1 expression in the liver. Notably, FZHY exerted a similar effect on ductular reaction and liver fibrosis as the Gli1 inhibitor GANT61 in CCl4/2-AAF-induced rats. In vitro, the inhibitory effect of FZHY on the differentiation of WB-F344 cells into a biliary phenotype was similar to the Gli1 inhibitor GANT61. Furthermore, Gli1 overexpression lentiviral transfection experiments confirmed that FZHY inhibited the differentiation of WB-F344 cells into a ductular phenotype in a Gli1-dependent manner. CONCLUSIONS:FZHY exerts a remarkable antifibrotic effect, which may be mediated by inhibiting the differentiation of HPCs into a ductular phenotype, potentially through suppression of the Gli1 pathway. This finding provides further insight into the mechanism of FZHY and suggests a potential role of Gli1 signaling in its antifibrotic action.

[This corrects the article DOI: 10.3389/fphar.2021.671152.].

BACKGROUND:Metabolic dysfunction-associated steatohepatitis (MASH) has emerged as a worldwide health challenge with few therapeutic options. Xiayuxue Decoction (XYXD), a classical herbal formula from the Synopsis of the Golden Chamber (Jin Gui Yao Lue), a classic by Zhang Zhongjing, comprises Prunus persica (Linn.) Batsch, Rheum palmatumLinn., and Eupolyphaga sinensis Walker. While clinically employed for the treatment of chronic liver diseases, including MASH, its precise molecular mechanisms remain undefined. AIM OF THE STUDY:This study aims to clarify the therapeutic mechanisms underlying the effects of XYXD in MASH, with a particular focus on investigating its roles in gut microbiota remodeling, bile acid (BA) metabolism, N6-methyladenosine (m6A) transcriptional modification, and arachidonic acid (AA) metabolism. METHODS:A MASH model was induced by using a methionine-choline-deficient (MCD) diet, and the therapeutic effect of XYXD was evaluated by analyzing lipid profiles, liver function parameters, and histopathological changes. Gut microbiota composition was characterized via 16S rRNA gene sequencing. Meanwhile, the metabolomic profiling of BA metabolites in the liver, serum, and feces, as well as AA derivatives in the liver, was performed by using LC-MS/MS. Additionally, the expression profiles of relevant mRNAs and proteins, including those related to BA metabolism, lipid homeostasis, inflammatory response, and m6A modification, were determined. Deoxycholic acid (DCA) and XYXD-containing serum were used to treat RAW264.7 macrophage cells to verify further their regulatory effects on inflammation, m6A modification, and AA metabolism in vitro. RESULTS:XYXD exhibits therapeutic efficacy against MASH through the dual regulation of inflammatory pathways and lipid metabolic homeostasis. It effectively reverses MCD diet-induced microbiota imbalance and maintains BA homeostasis by activating the farnesoid X receptor (FXR)-small heterodimer partner (SHP) pathway, with a particular role in reducing Clostridium abundance and DCA levels. Further investigations revealed that DCA mediates the upregulation of methyltransferase-like 13/14 mRNA, which in turn enhances m6A modification and influences AA metabolism. This integrated regulation of inflammatory, metabolic, and epigenetic pathways underscores XYXD's systemic therapeutic potential. CONCLUSION:XYXD alleviates MASH via the following multifaceted regulatory mechanism: it modulates gut microbiota dynamics, activates the FXR-SHP axis to maintain BA homeostasis, and ultimately regulates m6A transcriptional modification to influence AA metabolism. This coordinated network establishes functional crosstalk between microbiota and metabolic pathways in disease intervention.

BACKGROUND:MASH has become the most prevalent chronic liver disease worldwide. Hepatocyte ferroptosis is a key driver in the progression of MASH. Salvianolic acid B (Sal B), a natural antioxidant active ingredient, has an anti-MASH effect. However, the underlying mechanism by which Sal B inhibits hepatocyte ferroptosis and improves MASH remains unclear. METHODS:An in vivo MASH model was established by feeding C57BL/6J mice a HFHC diet for 30 weeks. In the 24th week, mice were treated with Sal B, obeticholic acid (OCA) or SRT1720. In vitro, over-expression and knockdown of SIRT1 in hepatocytes were constructed with lentivirus, subsequently, a hepatocyte lipid deposition model induced by palmitic acid/oleic acid (PO) or a hepatocyte ferroptosis model induced by erastin were used to investigate the efficacy and mechanism of Sal B. Furthermore, the interaction and binding kinetics of Sal B and SIRT1 were determined by molecular docking simulation, DARTS, CETSA and SPR assays. RESULTS:Sal B attenuated hepatic inflammation, lipid deposition and fibrosis in MASH mice. In situ transmission electron microscopy revealed characteristic morphological features of hepatocyte ferroptosis in the livers of MASH mice, which were alleviated by Sal B treatment. Mechanistically, Sal B up-regulated hepatic SIRT1 protein and inhibited p53 acetylation. In vitro, Sal B alleviated PO- or erastin-induced lipid deposition and ferroptosis in hepatocytes, along with up-regulating SIRT1 and inhibiting p53 acetylation. Similar protective effects were observed upon SIRT1 overexpression. Strikingly, SIRT1 knockdown reduced the inhibitory effect of Sal B on hepatocyte ferroptosis. SPR analysis confirmed moderate-affinity binding of Sal B to recombinant human SIRT1 protein, while DARTS and CETSA assays indicated that Sal B enhances SIRT1 enzymatic and thermal stability. CONCLUSION:Sal B ameliorates MASH in mice via directly regulating SIRT1/p53 signaling to suppress hepatocyte ferroptosis. Targeting hepatocyte ferroptosis through SIRT1 activation is a promising therapeutic strategy for MASH.

Cholestatic liver disease (CLD) is a severe hepatobiliary disorder with limited treatment options. Although the natural compound Isoastragaloside II (IAS II) has been suggested to possess general hepatoprotective properties, its therapeutic efficacy against CLD has not been reported. In this study, the therapeutic potential and underlying mechanisms of IAS II for CLD were investigated using a murine model established by 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) feeding. It was demonstrated that DDC-induced cholestatic liver injury was significantly alleviated by IAS II treatment, as indicated by the normalization of key serum biochemical parameters. An integrated analysis of histopathological, protein, and gene expression data demonstrated that IAS II treatment effectively suppressed the ductular reaction, inhibited hepatic fibrosis, and attenuated inflammation. Analysis of 16S rRNA sequencing data revealed that IAS II treatment was accompanied by alterations in gut microbiota composition, including an enrichment of Bacteroides and a reduction in Lachnospiraceae_NK4A136_group. Antibiotic depletion experiments demonstrated that the hepatoprotective effects of IAS II were abrogated by gut microbiota depletion, confirming the essential role of the gut microbiota in its pharmacological action. At the molecular level, IAS II activated both the PPAR-α pathway (as indicated by transcriptomic analysis) and the FXR pathway (as confirmed by subsequent validation), which collectively resulted in a significant reduction in the hepatic accumulation of toxic bile acids (BAs). In vitro studies in LCA-induced HepG2 cells further revealed that the restoration of PPAR-α, ACOX2, FXR, and BSEP protein expression by IAS II required receptor activation, as these effects were blocked by specific inhibitors. In summary, to our knowledge, this study provides the first demonstration that a protective effect against DDC-induced cholestatic liver injury is conferred by IAS II through coordinated regulation of PPAR-α and FXR signaling, restoration of BA homeostasis, and in a gut microbiota-dependent manner, with these effects requiring the activation of both PPAR-α and FXR pathways, thereby identifying IAS II as a novel and promising therapeutic candidate for CLD.

Abstract Background Fuzheng Huayu formula (FZHY) has been extensively applied in clinical for liver fibrosis treatment in China, its therapeutic potential in cholestatic liver injury remains underexplored. Objective To evaluate the protective effects and underlying mechanisms of FZHY against chronic cholestatic liver injury. Methods The therapeutic effect of FZHY was initially validated in a 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-induced murine model of chronic cholestasis. Subsequent mechanistic investigations were conducted through comparative analyses in peroxisome proliferator-activated receptor α gene knockout (Pparα −/−) mice subjected to DDC challenge. Results FZHY significantly ameliorated chronic cholestatic liver injury phenotypes in DDC-induced mice, as evidenced by bile acids (BAs) accumulation, inflammation, ductular reaction and biliary fibrosis was remarkably reduced after treatment with FZHY. Transcriptome sequencing analysis revealed that the effect of FZHY on chronic cholestatic liver injury was closely associated with activating PPAR signaling pathway and suppressing nuclear factor kappa-B (NF-κB) signaling. Further research found FZHY did not only enhance the total hepatic content of PPARα protein, but also increased its nuclear to cytoplasmic ratio that was reduced by DDC inducing. Additionally, FZHY suppressed hepatic phosphorylation of IκBα and NF-κB. The therapeutic effect of FZHY in treating DDC-induced mice with chronic cholestatic liver injury is similar to that of fenofibrate, a PPARα agonist. Crucially, genetic ablation of Pparα substantially abrogated the hepatoprotective and anti-fibrotic effects of FZHY in DDC-induced mice. Conclusions The present study underscores FZHY regulated BAs metabolism and alleviated hepatic inflammation and fibrosis by upregulating PPARa in DDC-induced mice. Our study provides novel insights that FZHY might be a promising drug for chronic cholestatic liver injury. Graphical Abstract

BackgroundDespite our previous findings that both total astragalus saponins (TAS) and hepatic AAV8.Numb-Exon3 overexpression (AAV8.Numb-Ex3OE) are effective against cholestatic liver fibrosis (CLF), the synergistic effect and mechanism of their combination remain unknown. This study was designed to investigate this combination therapy and elucidate its underlying molecular mechanisms.MethodsA rat model of CLF was induced by bile duct ligation and subsequently treated with combination therapy. Evaluations included serum biochemical parameters, liver histopathology, hepatic hydroxyproline (Hyp) content, expression levels of key molecules in the Notch signaling pathway, hepatic progenitor cells (HPCs) differentiation, ductular reaction (DR), and liver regeneration. In vitro, WB-F344 cells were induced toward a cholangiocyte-like phenotype, and the influence of TAS on HPCs differentiation was assessed following LV.Numb-Ex3 transfection.ResultsThe combination therapy more effectively improved serum biochemistry, reduced hepatic inflammation and collagen deposition, and lowered hepatic Hyp content compared to TAS or AAV8.Numb-Ex3OE monotherapy. Mechanistically, the combination therapy further suppressed differentiation of HPCs into cholangiocytes and inhibited DR, while promoting HPC-to-hepatocyte differentiation and enhancing liver regeneration.ConclusionThe combination of TAS and AAV8.Numb-Ex3OE exerts a synergistic anti-fibrotic effect in CLF by redirecting HPCs differentiation fate. These findings provide experimental support for the development of novel combinatorial strategies against liver fibrosis.

Rebleeding is a severe complication following recovery from esophageal variceal bleeding (EVB), yet robust predictive tools for assessing post-treatment risk after endoscopic variceal ligation (EVL) therapy remain scarce. This study developed and independently validated a machine learning (ML) model using multidimensional clinical data to predict 1-year rebleeding risk. Two independent cohorts were included: a retrospective cohort (n = 373) for model development and a prospective cohort (n = 119) for validation, with a one-year rebleeding endpoint. Predictors were identified using Recursive Feature Elimination (RFE), and eight ML algorithms were evaluated. Each algorithm was optimized via 5-fold cross-validation. The model with optimal performance was chosen to develop an online computational platform. RFE identified eight key predictors. The XGBoost model demonstrated superior predictive performance in both the training and validation cohorts, achieving AUCs of 0.883 and 0.887, respectively. This model was subsequently implemented in an online clinical platform for individualized 1-year rebleeding risk assessment. Our findings establish XGBoost as an effective tool for predicting EVB rebleeding risk, providing an evidence-based decision aid for post-EVL management.

BACKGROUND Rebleeding after recovery from esophagogastric variceal bleeding (EGVB) is a severe complication that is associated with high rates of both incidence and mortality. Despite its clinical importance, recognized prognostic models that can effectively predict esophagogastric variceal rebleeding in patients with liver cirrhosis are lacking. AIM To construct and externally validate a reliable prognostic model for predicting the occurrence of esophagogastric variceal rebleeding. METHODS This study included 477 EGVB patients across 2 cohorts: The derivation cohort (n = 322) and the validation cohort (n = 155). The primary outcome was rebleeding events within 1 year. The least absolute shrinkage and selection operator was applied for predictor selection, and multivariate Cox regression analysis was used to construct the prognostic model. Internal validation was performed with bootstrap resampling. We assessed the discrimination, calibration and accuracy of the model, and performed patient risk stratification. RESULTS Six predictors, including albumin and aspartate aminotransferase concentrations, white blood cell count, and the presence of ascites, portal vein thrombosis, and bleeding signs, were selected for the rebleeding event prediction following endoscopic treatment (REPET) model. In predicting rebleeding within 1 year, the REPET model exhibited a concordance index of 0.775 and a Brier score of 0.143 in the derivation cohort, alongside 0.862 and 0.127 in the validation cohort. Furthermore, the REPET model revealed a significant difference in rebleeding rates (P < 0.01) between low-risk patients and intermediate- to high-risk patients in both cohorts. CONCLUSION We constructed and validated a new prognostic model for variceal rebleeding with excellent predictive performance, which will improve the clinical management of rebleeding in EGVB patients.

BACKGROUND:Esophageal and gastric variceal bleeding is a catastrophic complication of portal hypertension, most commonly caused by cirrhosis of various etiologies. Although a considerable body of research has been conducted in this area, the complexity of the disease and the lack of standardized treatment strategies have led to fragmented findings, insufficient information, and a lack of systematic investigation. Bibliometric analysis can help clarify research trends, identify core topics, and reveal potential future directions. Therefore, this study aims to use bibliometric methods to conduct an in-depth exploration of research progress in this field, with the expectation of providing new insights for both clinical practice and scientific research. AIM:To evaluate research trends and advancements in esophagogastric variceal bleeding (EGVB) over the past twenty years. METHODS:Relevant publications on EGVB were retrieved from the Web of Science Core Collection. VOSviewer, Pajek, CiteSpace, and the bibliometrix package were then employed to perform bibliometric visualizations of publication volume, countries, institutions, journals, authors, keywords, and citation counts. RESULTS:The analysis focused on original research articles and review papers. From 2004 to 2023, a total of 2097 records on EGVB were retrieved. The number of relevant publications has increased significantly over the past two decades, especially in China and the United States. The leading contributors in this field, in terms of countries, institutions, authors, and journals, were China, Assistance Publique-Hôpitaux de Paris, Bosch Jaime, and World Journal of Gastroenterology, respectively. Core keywords in this field include portal hypertension, management, liver cirrhosis, risk, prevention, and diagnosis. Future research directions may focus on optimizing diagnostic methods, personalized treatment, and multidisciplinary collaboration. CONCLUSION:Using bibliometric methods, this study reveals the developmental trajectory and trends in research on EGVB, underscoring risk assessment and diagnostic optimization as the core areas of current focus. The study provides an innovative and systematic perspective for this field, indicating that future research could center on multidisciplinary collaboration, personalized treatment approaches, and the development of new diagnostic tools. Moreover, this work offers practical research directions for both the academic community and clinical practice, driving continued advancement in this domain.

ETHNOPHARMACOLOGICAL RELEVANCE:The traditional Chinese herb Rehmannia glutinosa (Gaertn.) DC. is an extensively employed to manage liver diseases. Rehmannioside D (RD), a major bioactive constituent of Rehmannia glutinosa (Gaertn.) DC, exhibits significant anti-apoptotic properties. Nevertheless, its efficacy and underlying mechanisms against hepatic fibrosis remain poorly defined. AIM OF THE STUDY:To evaluate the pharmacological effects of RD on liver fibrosis and elucidate its underlying mechanisms of action. MATERIALS AND METHODS:In vivo, the therapeutic efficacy of RD was evaluated in two classical liver fibrosis models: CCl4-treated mice and DMN-exposed rats. In vitro, RD's impact on hepatocyte apoptosis and the contribution of Sirtuin 7 (Sirt7) were investigated in H2O2-injured AML12 cells, using a selective Sirt7 inhibitor. RESULTS:RD treatment demonstrated a systematic reduction in hepatic injury, inflammation, and collagen deposition in vivo fibrosis models. RNA-sequencing analysis revealed that the therapeutic effects of RD was closely related to suppression of the p53 signaling pathway and cellular apoptosis. Similarly, RD significantly alleviated hepatocyte apoptosis in fibrotic livers, and this anti-apoptotic effect was mediated through the Sirt7/p53 signaling axis. In vitro, RD alleviated H2O2-induced apoptosis in AML12 cells, however, this protective effect was partially abrogated by pharmacological inhibition of Sirt7. CONCLUSION:Collectively, these findings indicated that RD attenuates hepatic fibrosis by upregulating Sirt7 expression, thereby inhibiting acetyl-p53-mediated hepatocyte apoptosis. Collectively, our findings establish RD as a promising anti-fibrotic agent and identify the Sirt7/p53 axis as its key mechanistic conduit.

Bone marrow-derived macrophages (BMDMs) regulate hepatic progenitor cells (HPCs) differentiation, potentially via the Wnt signaling pathway. While M1-polarized BMDMs (M1-BMDMs) exert anti-fibrotic effects in the liver, Wnt5a is implicated in fibrosis progression. The specific influence of Wnt5a levels within M1-BMDMs on HPCs fate and cirrhosis development remains unclear. This study aimed to elucidate the relationship between M1-BMDM-derived Wnt5a and HPCs differentiation during cirrhosis progression. First, Wnt5a protein expression was assessed in liver biopsy tissues from patients with hepatitis B-associated liver fibrosis. Second, cirrhosis was induced in rats using CCl4/2-AAF. In week 9, rats received intravenous injections of M1-BMDMs with Wnt5a knockdown (M1-BMDM Wnt5a−KD) or overexpression (M1-BMDM Wnt5a−OE); peripheral BMDMs recruitment was blocked using a CCR2 inhibitor. Fibrosis progression, ductular reaction (DR), and HPC differentiation were evaluated. In vitro, WB-F344 cells subjected to frizzled 2 (Fzd2) knockdown (WB-F344 Fzd2−KD) or overexpression (WB-F344 Fzd2−OE) were cultured with conditioned medium from M1-BMDM Wnt5a−KD (CM Wnt5a−KD) or M1-BMDM Wnt5a−OE (CM Wnt5a−OE). In patients with hepatitis B-related fibrosis, hepatic Wnt5a expression increased progressively with METAVIR fibrosis grade. In the rat cirrhosis model, M1-BMDMs Wnt5a−KD attenuated fibrosis, whereas M1-BMDMs Wnt5a−OE exacerbated it. Mechanistically, in vivo injection of M1-BMDMs Wnt5a−KD significantly inhibited HPCs differentiation into biliary epithelial cells (BECs), while M1-BMDM Wnt5a−OE promoted this differentiation. In vitro, CM Wnt5a−KD inhibited the differentiation of WB-F344 cells into BECs; this inhibition was potentiated by Fzd2 knockdown in WB-F344 cells but abrogated by Fzd2 overexpression. Conversely, under CM Wnt5a−OE conditions, WB-F344 Fzd2−OE cells exhibited increased cholangiocytic differentiation, an effect largely negated by Fzd2 knockdown. M1-BMDMs Wnt5a−KD demonstrated superior therapeutic efficacy against cirrhosis compared to unmodified M1-BMDMs. Wnt5a/Fzd2 signaling mediated the crosstalk between M1-BMDMs Wnt5a−KD and HPCs, revealing a novel therapeutic target for cirrhosis treatment.

Ethnopharmacological relevance Astragali Radix (Huangqi), a traditional Chinese medicine documented in Shennong Bencao Jing (ca. 200 AD), has been used for chronic liver disease. Total astragalus saponins (TAS), its primary bioactive component, exhibit anti-fibrotic effects. However, their potential synergy with stem cell therapy remains unexplored. Aim of the study To investigate whether TAS enhances the therapeutic efficacy of Numb-overexpressing human umbilical cord-mesenchymal stem cells (hUC-MSCs Numb-OE) against cholestatic liver fibrosis (CLF) and elucidate underlying mechanisms. Materials and methods CLF was induced in rats by bile duct ligation, post-surgery, rats received hUC-MSCs Numb-OE via intrasplenic injection. Starting the second week, TAS was administered daily by gavage for three weeks. Serum biochemistry, liver histopathology, hepatic hydroxyproline content, ductular reaction (DR), liver regeneration and Numb-p53 axis were assessed. In vitro, the effect of TAS on hUC-MSCs Numb-OE differentiation was evaluated. Results Combined TAS and hUC-MSCs Numb-OE significantly improved inflammatory response and cholestasis, reduced collagen deposition, and inhibited hepatic stellate cells activation and DR. Furthermore, the combination promoted differentiation of hUC-MSCs Numb-OE into hepatocytes confirmed by enhanced expression of hepatocyte nuclear factor 4α and albumin, and by EGFP (marked hUC-MSCs)/albumin immunofluorescent co-staining in vitro. Mechanistically, the combined therapy enhanced stability of the Numb-p53 axis demonstrated by increased expression levels of Numb and p53, alongside reduced expression levels of pNumb and Mdm2. Conclusions TAS synergistically enhances the anti-CLF efficacy of hUC-MSCs Numb-OE. This effect is associated with enhanced suppression of DR and promotion of liver regeneration, may mediated through stabilization of the Numb-p53 axis.

Cholestasis can occur in various acute and chronic liver diseases, with primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) being the most common clinical manifestations. Without appropriate treatment, these conditions may ultimately progress to liver cirrhosis and hepatic failure. Therefore, identifying novel therapeutic targets is of great importance. Our previous research had found a gene target named Numb which is a determinant of stem cell fate can increase the anti-cholestatic liver fibrosis (CLF) effect of bone marrow mesenchymal stem cells (BM-MSCs). However, whether the Numb gene or its exon has direct anti-CLF activity is unclear. In this study, an adeno-associated virus (AAV) was used as a gene delivery vector to overexpress the full-length Numb gene directly in the rat liver. In addition, Exon3 was overexpressed to clarify the effective site of Numb gene for comparison. AAV.Numb can alleviate CLF and suppressed the activation of Notch signaling and the differentiation of hepatic progenitor cells (HPCs) into biliary epithelial cells (BECs), and the anti-CLF effect of Numb-Exon3 was similar to that of full-length Numb. The findings revealed that Numb gene may be a new therapeutic target for PBC and that Exon3 may be an effective site.

The effectiveness and risks of anticoagulant therapy in cirrhotic patients with non-symptomatic portal vein thrombosis (PVT) remain unclear. We conducted a multicenter, Zelen-designed randomized controlled trial to determine the effectiveness of warfarin in cirrhotic patients with non-symptomatic PVT during a one-year follow-up. In brief, 64 patients were 1:1 randomly divided into the anticoagulation group or the untreated group. The probability of recanalization was significantly higher in the anticoagulation group than those untreated in both ITT analysis (71.9% vs 34.4%, p = 0.004) and PP analysis (76.7% vs 32.4%, p < 0.001). Anticoagulation treatment was the independent predictor of recanalization (HR 2.776, 95%CI 1.307-5.893, p = 0.008). The risk of bleeding events and mortality were not significantly different. A significantly higher incidence of ascites aggravation was observed in the untreated group (3.3% vs 26.5%, p = 0.015). In conclusion, warfarin was proved to be an effective and safe as an anticoagulation therapy for treating non-symptomatic PVT in cirrhotic patients.

ETHNOPHARMACOLOGICAL RELEVANCE:Chronic liver diseases mainly include chronic viral liver disease, metabolic liver disease, cholestatic liver disease (CLD), autoimmune liver disease, and liver fibrosis or cirrhosis. Notably, the compound formulas of traditional Chinese medicine (TCM) is effective for chronic liver diseases in clinical trials and basic research in vivo, which provide evidence of chronic liver disease treatment with integrated TCM and traditional Western medicine.AIM OF THE REVIEW:This paper aims to provide a comprehensive review of the compound formulas of TCM for treating different chronic liver diseases to elucidate the composition, main curative effects, and mechanisms of these formulas and research methods.MATERIALS AND METHODS:Different keywords related to chronic liver diseases and keywords related to the compound formulas of TCM were used to search the literature. PubMed, Scopus, Web of Science, and CNKI were searched to screen out original articles about the compound formulas of TCM related to the treatment of chronic liver diseases, mainly including clinical trials and basic in vivo research related to Chinese patent drugs, classic prescriptions, proven prescriptions, and hospital preparations. We excluded review articles, meta-analysis articles, in vitro experiments, articles about TCM monomers, articles about single-medicine extracts, and articles with incomplete or uncertain description of prescription composition. Plant names were checked with MPNS (http://mpns.kew.org).RESULTS:In this review, the clinical efficacy and mechanism of compound formulas of TCM were summarized for the treatment of chronic viral hepatitis, nonalcoholic fatty liver disease, CLD, and liver fibrosis or cirrhosis developed from these diseases and other chronic liver diseases. For each clinical trial and basic research in vivo, this review provides a detailed record of the specific composition of the compound formulas of TCM, type of clinical research, modeling method of animal experiments, grouping methods, medication administration, main efficacy, and mechanisms.CONCLUSION:The general development process of chronic liver disease can be summarized as chronic hepatitis, liver fibrosis or cirrhosis, and hepatocellular carcinoma. The compound formulas of TCM have some applications in these stages of chronic liver diseases. Owing to the continuous progress of medical technology, the benefits of the compound formulas of TCM in the treatment of chronic liver diseases are constantly changing and developing.

Total astragalus saponins (TAS) are the main active components of astragali radix, and have potent anti-hepatic fibrosis effect. However, the therapeutic efficacy of TAS and their potential mechanisms in the treatment of primary sclerosing cholangitis (PSC) remain unclear. In this study, two mouse models of PSC, including 3,5-Diethoxycarbonyl-1,4-Dihydro-2,4,6-Collidine (DDC)-induced PSC and Mdr2-/- spontaneous PSC, and the Tgr5-/- mice were used to investigate the therapeutic effect and mechanisms of TAS. Treatment with TAS, particularly with a dose of 56 mg/kg, significantly ameliorated the PSC-related liver injury, cholestasis, collagen deposition, ductular reaction (DR), and fibrosis in the DDC-induced and Mdr2-/-spontaneous PSC mice. Furthermore, treatment with TAS significantly mitigated the PSC-related inflammatory responses in vivo and HIBEpiC cells by inhibiting the expression of TNF-α, IL-6, and IL-1β. Mechanistically, treatment with TAS rescued the PSC-decreased hepatic TGR5 expression to attenuate the NF-κB p65 phosphorylation. Notably, the therapeutic efficacy of TAS on PSC in DDC-induced mice was abrogated in Tgr5-/- mice, suggesting the anti-PSC effect of TAS may depend on enhancing TGR5 expression. In conclusion, TAS ameliorated DR, inflammation and liver fibrosis in both models of PSC mice by rescuing TGR5 expression. Our findings may aid in the design of new therapeutic strategies for the treatment of PSC.

ETHNOPHARMACOLOGICAL RELEVANCE:Cholestatic liver diseases (CLD) are liver disorders resulting from abnormal bile formation, secretion, and excretion from various causes. Due to the lack of suitable and safe medications, liver transplantation is the ultimate treatment for CLD patients. Isoastragaloside I (IAS I) is one of the main saponin found in Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge, which has been demonstrated to obviously alleviate CLD. Nevertheless, the IAS I's specific anti-CLD mechanism remains undecipherable. AIM OF THE STUDY:This study's purpose was to elucidate the protective consequence of IAS I on 0.1% 3, 5-diethoxycarbonyl-1,4-dihydroxychollidine (DDC) diet-induced CLD mice, and to reveal its potential mechanism. MATERIALS AND METHODS:In this study, mice with CLD that had been fed a 0.1% DDC diet were distributed two doses of IAS I (20 mg/kg, 50 mg/kg). The effects of IAS I on CLD models were investigated by assessing blood biochemistry, liver histology, and Hyp concentrations. We investigated markers of liver fibrosis and ductular reaction using immunohistochemistry, Western blot, and qRT-PCR. Liver inflammation indicators, arachidonic acid (ARA), and ω-3 fatty acid (FA) metabolites were also analyzed. Quantitative determination of 39 bile acids (BAs) in different organs employing UHPLC-Q-Exactive Orbitrap HRMS technology. Additionally, the H&E and Western blot analysis were used to evaluate differences in intestinal barrier function in DDC-induced mice before and after administering IAS I. RESULTS:After treatment with IAS I, serum biochemical indicators and liver hydroxyproline (Hyp) increased in a dose-dependent manner in CLD mice. The IAS I group showed significant improvement in indicators of liver fibrosis and ductular response, including as α-smooth muscle actin (α-SMA) and cytokeratin 19 (CK19), and transforming growth factor-β (TGF-β)/Smads signaling pathway. And inflammatory factors: F4/80, tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β), ARA and ω-3 FA metabolites showed significant improvement following IAS I treatment. Moreover, IAS I significantly ameliorated liver tau-BAs levels, particularly TCA, THCA, THDCA, TCDCA, and TDCA contents, which were associated with enhanced expression of hepatic farnesoid X receptor (FXR), small heterodimer partner (SHP), cholesterol 7α-hydroxylase (Cyp7a1), and bile-salt export pump (BSEP). Furthermore, IAS I significantly improved pathological changes and protein expression related to intestinal barrier function, including zonula occludens protein 1 (ZO-1), Muc2, and Occludin. CONCLUSIONS:IAS I alleviated cholestatic liver injury, relieved inflammation, improved the altered tau-BAs metabolism and restored intestinal barrier function to protect against DDC-induced cholestatic liver diseases.