芪蛭益肺颗粒药物血清对TGF-β_1刺激下肺成纤维细胞Smad3、Smad4、Smad7mRNA表达的影响

Influences of Qizhi Yifei Capsule medicated serum on expressions of Smad3 mRNA,Smad4 mRNA and Smad7 mRNA in lung fibroblasts stimulated by TGF-β_1

目的明确芪蛭益肺颗粒药物血清对正常大鼠肺成纤维细胞转化生长因子-β1／Smads（TGF-β1／Smads）信号转导通路的调控作用。方法分离自出生2～3d大鼠肺组织并培养传代至第4代成纤维细胞，随机分为空白血清组、模型组和药物血清组。模型组、药物血清组先滴加含2．5μg/L浓度TGF-β1的DMEM，空白血清组滴加新鲜无血清DMEM；之后空白血清组、模型组滴加含5％浓度空白血清的DMEM，药物血清组滴入含5％芪蛭益肺颗粒药物血清的DMEM。培养48、72h后采用实时荧光PCR检验方法，检测各组细胞中Smad3、Smad4、Smad7的mRNA表达。结果模型组48h细胞中Smad3、Smad4mRNA表达升高，72h细胞中Smad3、Smad4、Smad7mRNA表达升高，与空白组比较差异均具有统计学意义（P〈0．05）。药物血清组48h细胞中Smad3、Smad4mRNA表达下降，Smad7mRNA表达升高，72h细胞中Smad3、Smad4mRNA表达下降，与模型组比差异均具有统计学意义（P〈0．05）。结论芪蛭益肺颗粒可能通过调节TGF-β1／Smads信号转导通路的传导，抑制TGF-β1刺激下的肺成纤维细胞增殖。

Objective To definite the regulation effect of Qizhi Yifei Capsule medicated serum on signal transduction pathway of TGF-β1Smads of fibroblasts in normal rats. Methods The fibroblasts were separated from lung tissue in rats after birth for 2-3 days, and then randomly divided into blank group, model group and medicated serum group. Firstly model group and medicated serum group were given DMEM （containing 2. 5 μg/L TGF-β1 ） and blank group was given fresh non-serum DMEM, and secondly blank group and model group were given DMEM with blank serum （5 % ） and medicated serum group was given DMEM with Qizhi Yifei Capsule medicated serum （5%）. The expressions of Smad3 mRNA, Smad4 mRNA and Smad7 mRNA were detected by using real-time fluorescence quantitative polymerase chain reaction （RT-PCR） after 48 h and 72 h. Results In model group, the expressions of Smad3 mRNA and Smad4 mRNA increased after 48 h, and those of Smad3 mRNA, Smad4 mRNA andSmad7 mRNA increased after 72 h compared with blank group （P 〈 0.05 ）. In medicated serum group, the expressions of Smad3 mRNA and Smad4 mRNA decreased and expression of Smad7 mRNA increased after 48 h, and the expressions of Smad3 mRNA and Smad4 mRNA decreased after 72 h compared with model group （P 〈 0.05 ）. Conclusion Qizhi Yifei Capsule can inhibit the proliferation of lung fibro- blasts stimulated by TGF-β1 through regulating the conduction of signal transduction pathway of TGF-β1 Smads.