甘参复方对大鼠肝星状细胞增殖和胶原合成的影响  

Effects of Ganshen Compound on Proliferation and Collagen Synthesis of Rat Hepatic Stellate Cells

作　　者：张楚焌 孙天石 董一昕 张凡 李腾辉 王淑艳 于雪 杜庆红 李卫红 ZHANG Chujun;SUN Tianshi;DONG Yixin;ZHANG Fan;LI Tenghui;WANG Shuyan;YU Xue;DU Qinghong;LI Weihong(College of Chinese Medicine,Beijing University of Chinese Medicine,Beijing China 100029)

机构地区：[1]北京中医药大学中医学院

出　　处：《中医学报》2019年第5期959-962,共4页Acta Chinese Medicine

基　　金：国家自然科学基金项目(81273885);北京市自然科学基金青年基金项目(7144223);北京中医药大学校级课题项目(2017-JYB-JS003)

摘　　要：目的:观察甘参复方对大鼠肝星状细胞增殖和胶原合成的影响,并分析甘参复方抗肝纤维化的分子机制。方法:采用体外培养的永生化细胞株大鼠肝星状细胞(hepatic sellate cell-T6,HSC-T6)。将甘参复方稀释成6个不同的浓度梯度,采用CCK-8法检测各组细胞的增殖情况,筛选出甘参复方对HSC-T6的有效作用浓度。之后将细胞分为阴性对照组,甘参复方低浓度组、中浓度组、高浓度组。采用ELISA法检测各组细胞上清液中的Ⅰ型胶原蛋白(collagenⅠ,ColⅠ)水平。然后,应用转化生长因子-β1(transforming growth factor-β1,TGF-β1)(1 mg·L^-1)刺激HSC-T6,观察甘参复方对刺激后细胞的增殖和胶原合成的影响。结果:甘参复方的6个不同浓度对HSC-T6的增殖有明显抑制作用,且浓度为0.5μmol·L-1时抑制作用最明显(P<0.01)。ELISA结果显示,甘参复方能够降低细胞上清中的ColⅠ含量,且与CCK-8结果相吻合,在0.5μmol·L^-1时ColⅠ分泌最少。甘参复方能够抑制TGF-β1刺激后的HSC-T6增殖和胶原合成。结论:甘参复方能够显著抑制星状细胞的增殖以及ColⅠ的表达,这是甘参复方抗肝纤维化的作用机制之一。Objective:To observe the effects of Ganshen Compound on proliferation and collagen synthesis of rat hepatic stellate cells,and to analyze the molecular mechanism of Ganshen Compound on anti-hepatic fibrosis.Methods:The immortalized cell line of hepatic sellate cell-T6(HSC-T6) was cultured in vitro.The Ganshen Compound was diluted into 6 different concentration gradients.The proliferation of cells in each group was detected by CCK-8 method.The effective concentration of Ganshen Compound on HSC-T6 was screened out.After that,the cells were divided into negative control group,Ganshen Compound low concentration group,medium concentration group and high concentration group.The levels of collagenⅠ(Col I) in cell supernatants were detected by ELISA.Then,HSC-T6 was stimulated by transforming growth factor-β1(TGF-β1)(1 mg·L^-1),and the effects of Ganshen Compound on cell proliferation and collagen synthesis were observed.Results:The proliferation of HSC-T6 was inhibited by 6 different concentrations of Ganshen Compound,and the inhibition was most obvious when the concentration was 0.5 μmol·L-1(P<0.01).ELISA results showed that Ganshen Compound could reduce the content of collagen type I in cell supernatant,which was consistent with the results of CCK-8.Col I secreted least at 0.5 μmol·L^-1.Moreover,Ganshen Compound can inhibit the proliferation and collagen synthesis of HSC-T6 stimulated by TGF-β1.Conclusion:Ganshen Compound can significantly inhibit the proliferation of stellate cells and the expression of collagen type I,which is one of the mechanisms of anti-hepatic fibrosis of Ganshen Compound.

关 键 词：肝星状细胞 甘参复方 肝纤维化 胶原蛋白 大鼠 

分 类 号：R285.5[医药卫生—中药学]