Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss.Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathways

• 作者: LIU Yuan BI Yan-meng PAN Ting ZENG Ting MO Chan SUN Bing GAO Lei LYU Zhi-ping
• 来源期刊: 中国结合医学杂志（英文版）
• 年，卷（期）: 2022,（1）
• 所属期刊栏目: Original Articles
• 页码范围: 60-68
• 页数: 9页
• 语种: 英文
• DOI: 10.1007/s11655-021-3298-5

摘要

Objective:To investigate the molecular mechanism underlying the anti-hepatic fibrosis activity of ethyl acetate fraction Dicliptera chinensis (L.) Juss.(EDC) in human hepatic stellate cells (HSCs) in vitro and in a carbon tetrachloride (CCl4)-induced hepatic fibrosis mouse model in vivo.Methods:For in vitro study,HSCs were pre-treated with platelet-derived growth factor (10 ng/mL) for 2 h to ensure activation and treated with EDC for 24 h and 48 h,respectively.The effect of EDC on HSCs was assessed using cell counting kit-8 assay,EdU staining,transmission electron microscopy,immunofluorescence staining,and Westem blot,respectively.For in vivo experiments,mice were intraperitoneally injected with CCl4 (2 μ L/g,adjusted to a 25％ concentration in olive oil),3 times per week for 6 weeks,to develop a hepatic fibrosis model.Forty 8-week-old male C57BL/6 mice were divided into 4 groups using a random number table (n=10),including control,model,positive control and EDC treatment groups.Mice in the EDC and colchicine groups were intragastrically administered EDC(0.5 g/kg) or colchicine (0.2 mg/kg) once per day for 6 weeks.Mice in the control and model groups received an equal volume of saline.Biochemical assays and histological examinations were used to assess liver damage.Protein expression levels of α-smooth muscle actin (α-SMA) and microtubule-associated protein light chain 3B(LC3B) were measured by Western blot.Results:EDC reduced pathological damage associated with liver fibrosis,downregulated the expression of α-SMA and upregulated the expression of LC3B (P＜0.05),both in HSCs and the CCl4-induced liver fibrosis mouse model.The intervention of bafilomycin A1 and rapamycin in HSCs strongly supported the notion that inhibition of autophagy enhanced α-SMA protein expression levels(P＜0.01).The results also found that the levels of phosphoinositide (PI3K),p-PI3K,AKT,p-AKT,mammalian target of rapamycin (mTOR),p-mTOR,and p-p70S6K all decreased after EDC treatment (P＜0.05).Conclusion:EDC has anti-hepatic fibrosis activity by inducing autophagy and might be a potential drug to be further developed for human liver fibrosis therapy.