赶黄草总黄酮对活化的肝星状细胞 TGF-β1/Smads信号通路的影响

Effect of total flavonoids of Penthorum chinense Pursh on activated hepatic stellate cells based on TGF-β1/Smads signaling pathway

为研究赶黄草总黄酮对TGF-β1(transforming growth factor beta 1)活化的肝星状细胞的作用及可能的机理,采用TGF-β1诱导人肝星状细胞(hepatic stellate cell LX-2,HSC-LX-2)活化,给予不同浓度赶黄草总黄酮后,MTT法检测赶黄草总黄酮对活化后的LX-2增殖的影响,划痕实验检测细胞迁移率,胶原收缩实验检测胶原收缩情况,酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测细胞培养液中一型胶原(collagenⅠ,ColⅠ)及纤连蛋白(fibronectin,FN)等细胞外基质的沉积,进一步采用Western blot法检测TGF-β1/Smads信号通路相关蛋白Smad2、Smad3、p-Smad2、p-Smad3、Smad7的表达。结果显示,赶黄草浓度在为5、9、13 mg/L均可抑制TGF-β1活化后的LX-2的增殖和迁移,且13 mg/L时效果最显著(P<0.01);另外,赶黄草总黄酮可减少ColⅠ及FN等细胞外基质(extracellular matrix,ECM)的沉积(P<0.01),并抑制胶原收缩;Western blot结果显示,赶黄草总黄酮作用后Smad3、p-Smad2和p-Smad3的表达均显著减少(P<0.05),Smad7的表达明显增加。赶黄草总黄酮能明显抑制LX-2的增殖,减少ECM的分泌,其作用机制可能与与其抑制TGF-β1/Smads信号通路传导有关。

To investigate the effect and possible mechanism of total flavonoids of Penthorum chinense Pursh(TFPCP) on activated hepatic stellate cells,human hepatic stellate cell LX-2(HSC-LX-2) was cultured,TGF-β1 was used to active LX-2,MTT method was used to detect the effect of TFPCP on LX-2 proliferation rate,the cell migration rate was detected by scarification test and the collagen contraction was detected by collagen contraction test,enzyme-linked immunosorbent assay(ELISA) was adopted to detect the content of collagen I(Col I) and fibronectin(FN),Smad2,Smad3,p-Smad2,p-Smad3,Smad7 protein expressions in LX-2 was determined by Western blot.The results showed that the proliferation and migration of LX-2 were suppressed by TFPCP at three concentrations of 5,9 and 13 mg/L,and the effect was most significant at 13 mg/L(P<0.01).In addition,TFPCP could inhibit collagen contraction and reduce the deposition of Col I and FN(P<0.01).Further study indicated that treated with TFPCP suppressed the protein expressions of Smad3,phospho-Smad2 and phospho-Smad3(P<0.05),while significantly promoted the expressions of Smad7.In general,TFPCP inhibited the proliferation and migration of LX-2,reduced the secretion of ECM,the mechanism may be related to its inhibition of TGF-β1/Smads signaling pathway.