新疆假龙胆调控TGF-β1/Smad通路抑制肝星状细胞活化的作用机制

Mechanism of inhibition of hepatic stellate cell activation by TGF-β1/Smad pathway regulated by Gentianella turkestanorum

目的:探讨新疆假龙胆提取物通过调控转化生长因子β1(TGF-β1)/Smad通路发挥防治肝纤维化的具体作用机制。方法:采用TGF-β1(10 ng/mL)诱导LX-2细胞构建肝纤维化模型,对肝纤维化模型细胞给予不同浓度(100、200、300μg/mL)新疆假龙胆提取物,干预时间均为48 h,采用CCK8法检测干预后各组细胞的增殖抑制情况,流式细胞技术检测各组细胞的凋亡情况,荧光定量PCR和Western blot法检测各组Smad相关信号Smad3、Smad4、Smad7、TGF-β1 mRNA及蛋白表达。结果:与模型组比较,高、中浓度新疆假龙胆提取物可抑制TGF-β1诱导的LX-2细胞增殖与活化(P<0.05),上调细胞内Smad3、Smad4、TGF-β1 mRNA和蛋白表达(P<0.05),下调Smad7 mRNA和蛋白表达(P<0.05)。结论:新疆假龙胆提取物对TGF-β1诱导的LX-2细胞具有抑制增殖和促进凋亡的作用,并通过良性调控TGF-β1/Smad通路,从而延缓肝纤维化进程。

Objective:The experiment investigated the specific mechanism of action with Gentianella turkestanorum extract in preventing liver fibrosis by modulating the TGF-β1/Smad pathway.Methods:A liver fibrosis model was constructed using TGF-β1(10 ng/mL)stimulated LX-2 cells,and different concentrations(100,200,300μg/mL)of Gentianella turkestanorum extract were given to the liver fibrosis model cells,and the intervention time was 48 h.The proliferation inhibition of each group of cells after the intervention was detected by CCK8 method,and the apoptosis of each group of cells was detected by flow cytometry,fluorescence quantitative PCR and Western-blot assay were used to detect the mRNA and protein expression of Smad-related signals Smad3,Smad4,Smad7 and TGF-β1 pathway in each group.Results:The high and medium dose groups of Gentianella turkestanorum extract inhibited TGF-β1-induced LX-2 cell proliferation and activation,up-regulated intracellular mRNA and protein expression of Smad3,Smad4 and TGF-β1(P<0.05),and down-regulated mRNA and protein expression of Smad7(P<0.05).Conclusion:The extract of Gentianella turkestanorum had an inhibitory effect on TGF-β1-induced LX-2 cells by inhibiting proliferation and promoting apoptosis,and delayed the process of liver fibrosis by benignly regulating the TGF-β1/Smad pathway.