葫芦茶苷对大鼠肝星状细胞增殖和细胞外基质的影响及机制研究

Effects of Tadehaginoside on Rat Hepatic Stellate Cells Proliferation and Extracellular Matrix and Its Underlying Mechanism

目的研究葫芦茶苷对大鼠肝星状细胞增殖和细胞外基质分泌的影响,并探讨其可能机制。方法以大鼠肝星状细胞(HSC-T6)为体外细胞模型,采用CCK-8法检测葫芦茶苷对HSC-T6增殖的抑制作用,ELISA法测定细胞上清液中Hyp、HA和TIMP-1的含量,FQ-PCR检测Col I、TGF-β1、α-SMA、PDGF-BB mRNA的表达,Western blot法检测细胞内TGF-β1、α-SMA和PDGF-BB蛋白的表达。结果葫芦茶苷对HSC-T6细胞的增殖有较强的抑制作用。与空白组比较,葫芦茶苷各剂量组能降低细胞上清液中Hyp、HA和TIMP-1的含量,下调细胞内Col I、TGF-β1、α-SMA、PDGF-BB mRNA的表达,而且能降低细胞内TGF-β1、α-SMA和PDGF-BB蛋白的表达。结论葫芦茶苷体外具有抗肝纤维化作用,其机制可能是抑制HSC-T6细胞的增殖,下调TGF-β1、α-SMA和PDGF-BB蛋白的表达,从而减少肝细胞外基质的合成和沉积。

Objective To investigate the potential pharmacological activity of tadehaginoside(TA)on rat hepatic stellate cells proliferation and extracellular matrix and its underlying mechanism.Methods This experiment established a cell model using rat hepatic stellate cells(HSC-T6),HSC-T6 are cultured in vitro,and CCK-8 assay is used to evaluate the inhibitive effect of TA.The contents of Hyp,HA and TIMP-1 in HSC-T6 cell supernate are detected by ELISA.The relative quantification of Col I,TGF-β1,α-SMA,PDGF-BB mRNA expression in HSC-T6 cell is detected by FQ-PCR.The protein expression of TGF-β1,α-SMA,PDGF-BB related to HSC-LX2 cell cycle by Western Blotting.Results TA display strong inhibitory action to the proliferation of HSC-T6 cell.Compared with the blank group,the contents of Hyp,HA and TIMP-1 in cell supernate are decreased in each dose group of TA.In addition,TA could down-regulated the expression of Col I,TGF-β1,α-SMA,PDGF-BB mRNA,and the expressions of TGF-β1,α-SMA and PDGF-BB are all decreased.Conclusion TA has a anti-hepatic fibrosis effect in vitro,the mechanism may inhibit the proliferation of hsc-t6 cells,and downregulated expression of TGF-β1,α-SMA and PDGF-BB proteins,thus reduces the synthesis and deposition of the extracellular matrix.