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目的:观察化浊解毒含药血清对体外培养活化的大鼠肝星状细胞增殖及活性的影响,探讨化浊解毒方治疗肝纤维化的作用机制。方法:不同剂量(分别含有生药30,60,120 g·L-1)的化浊解毒方,按0.01 mL.g-1体重SD大鼠灌胃给药,以制备含药血清;常规培养活化的肝星状细胞(HSC-T6),用不同浓度的含药血清进行干预,用MTT,ELISA,RT-PCR法分别检测肝星状细胞增殖、细胞上清液中I型胶原的含量以及细胞中转化生长因子-β1(TGF-β1)mRNA基因的表达。结果:与正常血清组比较,化浊解毒方含药血清各浓度组均呈现出抑制HSC增殖的作用(P<0.05),且呈剂量和时间依赖性。与正常血清组比较,化浊解毒方含药血清各个浓度组在作用48 h后,细胞上清液中的I型胶原的含量均降低(P<0.05),均能下调细胞中TGF-β1mRNA的表达(P<0.05)。结论:化浊解毒方之所以能有效的治疗肝纤维化可能与化浊解毒方抑制肝星状细胞的增殖和活性有关。
Objective: To observe the effect of Huazhuo Jiedu decoction-containing serum on the proliferation and activity of cultured rat hepatic stellate cells in vitro and to explore the mechanism of Huazhuo Jiedu Decoction in treating hepatic fibrosis. Methods: Huazhuo Jiedu decoction of different doses (30,60,120 g · L-1 crude drug) was given intragastrically to SD rats of 0.01 mL.g-1 body weight for preparation of drug-containing serum. Conventional cultured activated liver Astrocytes (HSC-T6) were treated with serum containing different concentrations. The proliferation of hepatic stellate cells, the content of type I collagen in the supernatant of the cells and the transformation in the cells were detected by MTT, ELISA and RT-PCR Growth factor-β1 (TGF-β1) mRNA gene expression. Results: Compared with the normal serum group, the concentration of Huazhuo Jiedu Decoction serum showed inhibitory effects on the proliferation of HSC (P <0.05) in a dose-and time-dependent manner. Compared with the normal serum group, the content of type I collagen in the cell supernatant decreased (P <0.05) after 48 h treatment in all concentration groups of Huazhuo Jiedu Decoction serum, which could down-regulate the expression of TGF-β1 mRNA Expression (P <0.05). Conclusion: Huazhuo Jiedu Fang can effectively treat liver fibrosis and Huazhuo Jiedu Prescription inhibit hepatic stellate cell proliferation and activity.