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目的明确大黄灵仙胶囊对胆管内皮细胞TGF-β1mRNA、Smad2/3 mRNA表达的调节作用,阐明其是否能够逆转胆管内皮细胞纤维化进程,从而发挥临床疗效。方法体外培养人原代胆管内皮细胞株,以适当LPS给予刺激,采用大黄灵仙胶囊含药血清进行干预,细胞实验分为空白组、模型对照组、大黄灵仙胶囊大剂量组和小剂量组,在干预后24h、48h、72h提取细胞总RNA,采用荧光PCR技术,测定TGF-β1mRNA和Smad2/3 mRNA表达情况。结果大黄灵仙胶囊能降低TGF-β1mRNA表达水平,在含药血清干预处理后48h、72h与模型对照组比较,差异有统计学意义(P<0.05),且成剂量依赖性,同样在干预后48h、72h细胞内Smad2/3 mRNA表达明显降低,与对照组比较差异有统计学意义(P<0.05)。结论大黄灵仙胶囊对TGF-β1-Smad2/3信号转导通路具有显著的下调作用,其能缓解早期肝内胆管纤维化进程,达到防治肝内胆管结石形成的临床效果。
Objective To clarify the effect of rhubarb Lingxian capsule on the expression of TGF-β1mRNA and Smad2 / 3 mRNA in bile duct epithelial cells and to clarify whether it can reverse the process of fibrogenesis of bile duct endothelial cells and thus exert its clinical effect. Methods Human primary bile duct endothelial cell line was cultured in vitro and stimulated with appropriate LPS. The serum containing rhubarb and Lingxian capsule was used for intervention. The cell experiment was divided into blank group, model control group, rhubarb Lingxian capsule high dose group and low dose group The total RNA was extracted at 24h, 48h and 72h after intervention. The expression of TGF-β1mRNA and Smad2 / 3 mRNA were detected by fluorescence PCR. Results Rhubarb Lingxian capsule could decrease the expression of TGF-β1 mRNA, and there was a significant difference (P <0.05) between the model group and the control group at 48h and 72h after treatment with the drug-containing serum in a dose-dependent manner. The expression of Smad2 / 3 mRNA in 48h, 72h cells was significantly lower than that in control group (P <0.05). Conclusion Rhubarb Lingxian capsule has a significant down-regulation of TGF-β1-Smad2 / 3 signal transduction pathway, which can relieve the process of early intrahepatic bile duct fibrosis and achieve the clinical effect of preventing and treating intrahepatic bile duct stones.