扶正化瘀319方药物血清对肝星状细胞Ⅰ型胶原及转化生长因子β1表达的影响

目的：探讨扶正化瘀３１９方抗肝纤维化的主要作用机理。方法：分离正常大鼠肝星状细胞，并伟代培养。用扶正化阏９方给 正常大鼠灌胃，药物血清，温育培养细胞。胶原酶消化法测定细胞内外胶原生成率，ＥＬＩＳＡ 培养上清的Ｉ型胶原含量，免疫细胞化学染色、图像分析半定量转化生长因子β１（ＴＧＦβ１）蛋白表达，ＲＴ－ＰＣＲ法分析Ｉ型前胶原、ＴＧＦβ１ｍＲＮＡ的表达量。结果：扶正化瘀３１９方药物血清能明显抑制肝星状细

Serapharmacological effect of fuzheng huayu 319 Decoction on expression of type I collagen and transforming growth factor beta 1 in hepatic stellate cells]

OBJECTIVE: To investigate the main serapharmacological mechanism of antifibrotic action of Fuzheng Huayu 319 Decoction (FHD) at cellular and molecular level. METHODS: Hepatic stellate cells (HSC) were isolated from the normal rats and subcultured. Normal rats were medicated with FHD and their blood serum collected to prepare the medicated serum, which was incubated with the subcultured HSC. Then, the extra- and intracellular collagen synthetic rates were assayed with collagenase digestion method, type I collagen content in the medium measured with ELISA, transforming growth factor beta 1 (TGF beta 1) protein production semiquantified by immunocytochemistry stain and image analysis system, the gene expressions of type I procollagen and TGF beta 1 analyzed by RT-PCR. RESULTS: FHD medicated serum could obviously inhibit the extra- and intracellular collagen synthetic rates, decreased type I procollagen mRNA expression and its secretion, downregulate TGF beta 1 mRNA and its protein expression. CONCLUSION: FHD could inhibit HSC activation markedly, the main mechanism of its antifibrotic action may be the medicated serum inhibition on type I procollagen and TGF beta 1 gene expression.