Anti-inflammatory and Anti-apoptotic Effects of Hypericum Japonicum and its Active Compounds in Hepatic FibrosisOA

Objective:To explore the therapeutic efficacy of Hypericum japonicum(HJ)and its active compounds in mitigating hepatic fibrosis by elucidating their anti-inflammatory and anti-apoptotic effects.Methods:Twenty-four mice were randomly divided into 4 groups:control group,model group,HJ-1 mg/g group,and HJ-2 mg/g group.A hepatic fibrosis model was established via carbon tetrachloride(CCI_(4))injection at a dose of 5μL/g of body weight,twice weekly for 6 weeks.Subsequently,mice were gavaged with 1 mg/g or 2 mg/g HJ aqueous solution to assess its effects on liver pathological injury,liver function markers,and fibrosis markers in model mice.Integrated analyses employing network pharmacology,molecular docking,and eukaryotic transcriptomics were conducted to identify interactions between key HJ active constituents(e.g.,mairin,beta-sitosterol,quercetin,kaempferol)and core targets[e.g.,B-cell lymphoma-2(Bcl-2),tumor protein p53(TP53),tumor necrosis factor-alpha(TNF-α)],along with associated signaling pathways.Additionally,in vitro experiments were used to validate the effect of HJ active constituents on the viability of CCl4-injured HepG2 cells,expression of inflammatory cytokines,and apoptotic pathways.The study further utilized 16S rRNA amplicon sequencing to explore the modulatory effects of HJ on gut microbiota structure and function.Results:In vivo experiments demonstrated that HJ intervention significantly ameliorated liver pathological damage,reduced liver function markers,and downregulatedα-smooth muscle actin expression in mice(P<0.01).Network pharmacology and nolecular docking revealed significant binding affinity between key HJ active constituents and core targets(e.g.,Bcl-2,TP53,TNF-α).Eukaryotic transcriptomic analysis further identified core differentially expressed genes modulated by HJ,which were implicated in inflammatory response regulation and extracellular matrix receptor interaction pathways.Although HJ intervention did not significantly alter gut microbiota richness,it modulated metabolic and immune functions closely associated with hepatic fibrosis.In vitro,HJ active constituents significantly increased the viability of CCl4-injured HepG2 cells,downregulated TNF-αand interleukin-6 mRNA expression,and suppressed the mitochondrial apoptotic pathway via Bcl-2 regulation(P<0.01).Among the compounds,mairin demonstrated the most potent effect in reducing aspartate aminotransferase and hydroxyproline levels,while mairin and beta-sitosterol exerted anti-fibrotic effects by suppressing TNF-αand decreasing glutathione,respectively.Conclusions:HJ and its active ingredients elucidate the anti-inflammatory and anti-apoptotic effects in hepatic fibrosis.Further they emerge as promising candidates for targeted therapy against hepatic fibrosis.