摘要
目的观察中药复方861对体外培养肝星状细胞增殖及胶原合成的影响。方法分离培养大鼠肝星状细胞 ,传一代后加入不同浓度的复方861作用48小时 ,以MTT比色法检测对细胞增殖的作用。传一代细胞以10mg/ml复方861作用48小时后 ,以ELISA法、斑点杂交法检测胶原蛋白分泌及相应mRNA水平的变化。结果1.56～100mg/ml复方861(经0.45μm滤膜过滤)可使MTT转化率下降 ,细胞增殖受抑 ,呈明显的剂量依赖性 ;10mg/ml复方861可使I、III、IV型胶原及转化生长因子β1(TGFβ1)mRNA水平及相应蛋白分泌量下降。结论复方861抗纤维化的部分机理在于抑制活化肝星状细胞的增殖及胶原蛋白的合成 ,从而减少胶原纤维在肝脏内的沉积。
Aim The effect of compound 861(Cpd 861) on rat hepatic stellate cell(HSC) proliferation and collagen synthesis in vitro was observed. Methods The second passage of HSC was exposed to Cpd 861 at a concentration of 1.56～100 mg/ml for 48 hours. MTT colorimetric assay was used to evaluate the effect of Cpd 861 on HSC proliferation. After incubation with Cpd 861 for 48 hours, HSC was harvested to determine collagen type I,III,IV mRNA by dot blot hybridization and cell culture medium was used to detect collagen secretion by ELISA method. Results Cpd 861(through 0.45 μm filter membrane) could reduce MTT transforming rate and the inhibitory effect had a positive relation with the concentration of Cpd 861 in the cultured medium. Type I,III,IV collagen and TGFβ1 productions, were inhibited by Cpd 861 at a concentration of 10 mg/ml. Conclusion The antifibrotic mechanism of Cpd 861 was partly due to its inhibition of HSC proliferation and collagen production, which would reduce deposition in the liver.
基金
国家中医药管理局资助课题
