白花丹醌对瘦素刺激人肝星状细胞转化生长因子-β_1表达的影响

Effects of plumbagozeylanica(plumbago) on expression of transforming growth factor beta 1 in human hepatic stellate cells activated by leptin

目的研究白花丹醌对瘦素刺激体外培养人HSC-LX2TGF-β1 mRNA和蛋白表达的影响。方法体外培养HSC-LX2,瘦素(leptin)刺激24h,各药物与细胞共孵育24h后,采用荧光PCR检测各组TGF-β1 mRNA的表达和免疫组织化学方法检测TGF-β1蛋白表达情况。结果与leptin组比较,白花丹醌各剂量组作用24h后,HSC-LX2细胞中TGF-β1 mRNA的水平明显降低,尤以中、高剂量组明显(P<0.01);免疫组化结果显示白花丹醌各剂量组对HSC-LX2细胞TGF-β1蛋白表达有一定的抑制作用,且作用呈剂量依赖性。结论白花丹醌抗肝纤维化作用机制之一是从mRNA和蛋白水平抑制TGF-β1表达,从而抑制HSCECM的合成,发挥抗肝纤维化作用。

Aim To investigate the effect of plumbagozeylanica （plumbago） on the expression of TGF-β1 protein and TGF-β1mRNA in human hepatic stellate cells（HSC-LX2） activated by leptin.Methods HSC-LX2 were cultured in vitro,stimulated by leptin for 24 hours and treated with different concentrations of plumbago for 24 hours,The expressions of TGF-β1mRNA were determined by Real-time quantitative PCR using SYBR Green I,and the expressions of TGF-β1were determined by immunohistochemistry. Results The expressions of TGF-β1mRNA were significantly reduced by the treatment with three concentrations of plumbago than that in the leptin group,especially in middle and high dose group（P0.01）; the result of immunohistochemistry showed that expression of TGF-β1 in the plumbago-treatment groups was to some extent lower than that in the leptin group （P0.05）,and the inhibitory effect was in a dose-dependent manner.Conclusion The expressions of TGF-β1can be down-regulated by plumbago,resulting in the reduction of the synthesis of ECM and the promotion of the degradation of ECM,which may be one of the possible molecular mechanisms against hepatic fibrosis.