抗肝硬化复方在体外对星状细胞增殖和胶原合成的抑制作用

Inhibitory effect of BJ-JN on proliferation and collagen synthesis of HSC-T6 cells

目的研究抗肝硬化复方（BJ-JN）在体外对活化星状细胞（HSC）增殖和胶原合成的抑制作用，进一步探讨BJ-JN的作用机制。方法在引入血清药理方法的基础上，探讨BJ-JN在体外对混合新生小牛血清（NBS）＋大鼠血清（RS）和转化生长因子β1（TGF-β1）两种刺激因素作用下的HSC-T6细胞增殖和胶原合成的抑制作用。将不同时间点（2．5、3．5h）采集的BJ-JN含药血清作用于NBS和RS共同刺激下的HSC-T6，分别加入^3H-TdR和^3H-Pro，在液体闪烁计数仪上检测HST-T6细胞增殖和胶原合成。在5个不同的时间点（2～24h）将刺激浓度为9pmol／L的TGF-β1作用于HSC-T6，用四甲基偶氮唑（MTT）比色法测定吸光度值，确定TGF-β1刺激HST-T6的最适刺激时间。将不同浓度（0．19％～3．125％）BJ-JN含药血清作用于最适刺激时间、9pmol／L TGF-β1刺激条件下的HST-T6，用MTT比色法测定吸光度值并计算抑制率。结果与正常对照组比较，给药2．5和3．5h BJ-JN含药血清对NBS＋RS刺激的HSC-T6细胞增殖和胶原合成有显著的抑制作用。9pmol／LTGF-β1对HSC-T6细胞作用16h的刺激作用最强。不同浓度BJ-JN含药血清可显著抑制TGF-β1刺激的HSC-T6细胞增殖。结论BJ-JN含药血清在体外可以明显抑制HSC-T6细胞增殖和胶原合成。

Objective To study the inhibitory effect of BJ-JN on proliferation and collagen synthesis of active HSC cells in vrtro and its mechanism. Methods The inhibitory effect of BJ-JN on the proliferation and collagen synthesis of HSC-T6 cells was investigated, which was stimulated by mixed sera and transforming growth factor β1 respectively, by serum pharmacology method. The effect of serum containing BJ-JN in different time （2. 5 ,3.5 h） on the proliferation and collagen synthesis of HSC-T6 cells costimulated by mixed sera containing new bovine serum （NBS） and rat serum（RS） was measured by [^3H]-TdR and [^3H]-Proline incorporation assay. In additionally, the effect of TGF-β1（9 pmol/L） in different stimulus durations（2 -24 h）and the effect of drug-contained serum of BJ-JN in different concentrations（0. 19% -3. 125%） on proliferation of HSC-T6 stimulated by TGF-β1（9 pmol/L） in the optimal stimulus duration was measured by MTT assay. Results Drug-contained serum of BJ-JN in 2. 5 h and 3.5 h could markedly suppress proliferation and collagen synthesis of HSC-T6 cells stimulated by mixed sera. The optimal stimulus duration of TGF-β1 （9 pmol/L） on HSC-T6 was 16 h. Drug-contained serum of BJ-JN could markedly inhibit the activation and proliferation of HSC-T6 cells induced by TGF-β1（9 pmol/L）. Conclusion These results suggest that BJ-JN has inhibitory effect on proliferation and collagen synthesis of active HSC-T6 cells in vitro.