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目的应用基因芯片技术探讨甘草酸对大鼠肝星状细胞(HSC)转化生长因子(TGF)-β信号转导通路相关基因表达的作用。方法体外分离、培养大鼠HSC,分为对照组、TGF-β_1组(5ng/ml)、TGF-β_1(5ng/ml)+甘草酸(100μmol/L)组,10h后分别收集细胞抽提总RNA。应用针对于TGF-β/BMP信号转导通路的GEArray~(TM)Q基因芯片技术,筛选出甘草酸作用后HSC中TGF-β信号通路表达发生明显改变的相关基因。结果经TGF-β_1作用后上调,再经甘草酸作用后下凋的基因有16项。占16．7%(如Smad蛋白2,Smad蛋白3,Smad蛋白7,α1Ⅲ型前胶原,α2Ⅰ型前胶原,纤溶酶原激活物抑制剂1)；经TGF-β_1作用后下调,再经甘草酸作用后上调的基因有5项,占5．2%(如骨成型蛋白7,胰岛素生长因子结合蛋白3等)；经TGF-β_1作用后上调,再经甘草酸作用后上调更显著的基因有2项,占2．1%(转化生长因子2型受体,转化生长因子受体3)。并用RT-PCR法验证了部分基因(Smad蛋白2,Smad蛋白3,Smad蛋白7)mRNA的表达与基因芯片中变化趋势的一致性。结论甘草酸可能通过干预大鼠HSC中TGF-β信号通路,减少胶原合成,促进细胞外基质降解而发挥抗纤维化的分子机制。
Objective To investigate the effect of glycyrrhizin on the gene expression of transforming growth factor-β (TGF-β) in rat hepatic stellate cells (HSC) by gene chip technique. Methods Rat HSCs were isolated and cultured in vitro and divided into control group, TGF-β1 group (5ng / ml) and TGF-β1 (5ng / ml) + glycyrrhizinate group (100μmol / RNA. The GEArray ~ (TM) Q gene chip targeting to TGF-β / BMP signal transduction pathway was used to screen out the related genes that changed the expression of TGF-β signaling pathway in HSC after glycyrrhizin. Results After TGF-β 1 treatment, 16 genes were up-regulated after glycyrrhizic acid treatment. Accounting for 16.7% (such as Smad protein 2, Smad protein 3, Smad protein 7, α1 Ⅲ procollagen, α2 Ⅰ procollagen, plasminogen activator inhibitor 1); Glycyrrhizic acid increased after the role of genes 5, accounting for 5.2% (such as bone morphogenetic protein 7, insulin-binding protein 3, etc.); TGF-β_1 role after up-regulation, and then by glycyrrhizic acid increased more significantly There are 2 genes, accounting for 2.1% (transforming growth factor type 2 receptor, transforming growth factor receptor 3). RT-PCR method was used to verify the consistency of the changes of mRNA expression of some genes (Smad protein 2, Smad protein 3, Smad protein 7) with the gene chip. Conclusion Glycyrrhizic acid may exert its anti-fibrosis mechanism by interfering with TGF-β signaling pathway in rat HSC, reducing collagen synthesis and promoting extracellular matrix degradation.