芪蛭益肺药物血清对转化生长因子-β1刺激下肺成纤维细胞中基质金属蛋白酶及其抑制剂基因表达的影响

Influence of Qizhi Yifei Containing Serum on Regulating Expression of MMP-9 and TIMP-1 mRNA in Rat Ffibroblasts Stimulated by TGF-β1

目的观察芪蛭益肺药物血清对肺成纤维细胞中基质金属蛋白酶(MMP)-9及其抑制剂-1(TIMP-1)mRNA表达的影响,探讨其作用机制。方法采用胰酶消化法提取大鼠肺组织成纤维细胞,培养至第4代后随机分为空白血清组、模型组和药物血清组。模型组与药物血清组先滴加含0.0025μg/mL转化生长因子-β1(TGF-β1)的DMEM,空白血清组滴加新鲜无血清DMEM,随后空白血清组、模型组滴加含5%空白血清的DMEM,药物血清组加入5%含芪蛭益肺药物血清的DMEM。培养48、72 h后,采用行实时荧光定量PCR法检测各组细胞MMP-9、TIMP-1 mRNA表达。结果与空白血清组比较,培养48 h后模型组和药物血清组成纤维细胞中MMP-9、TIMP-1 mRNA表达上升,差异有统计学意义(P<0.05),药物血清组和模型组之间比较差异无统计学意义。与空白血清组比较,培养72 h后模型组MMP-9 mRNA表达上升;与模型组比较,培养72 h后药物血清组MMP-9 mRNA表达下降,差异均有统计学意义(P<0.05);TIMP-1 mRNA表达3组间比较差异无统计学意义。结论芪蛭益肺颗粒药物血清能够抑制TGF-β1刺激后肺成纤维细胞中MMP-9 mRNA高表达。

Objective To observe the influence of Qizhi Yifei containing serum on the expression of MMP-9 and TIMP-1 mRNA in lung fibroblasts, and explore its mechanism. Methods Trypsin digestion method was used to extract fibroblasts from lung tissue in rats. All fibroblasts were cultured and trained to the fourth generation. Then they were randomly divided into control group, model group and drug serum group. The model group and drug serum group were firstly treated by DMEM with 0.002 5 μg/mL TGF-β1. The control group was treated by DMEM only. The control group and model group were then treated by DMEM with blank drug serum in concentration of 5%, and the drug serum group was treated by DEME with Qiahi Yifei containing serum in same concentration. After 48 and 72 hours, RT-PCR was performed to test the expression of MMP-9 mRNA and TIMP-1 mRNA of each group. Results After 48 hours, MMP-9 and TIMP-1 mRNA were significantly increased in model group and drug serum group compared with control group. There was no difference between model group and drug serum group. After 72 hours, MMP-9 mRNA was up-regulated in model group and was decreased in drug serum group compared with control group. There was no significant difference among the three groups on the expression of TIMP-1 mRNA. Conclusion Qizhi Yifei containing serum can decrease the up-regulated expression of MMP-9 mRNA in lung fibroblasts stimulated by TGF-β1.