阿魏酸哌嗪对TGF-β_1诱导肾成纤维细胞表型活化的影响

Effects of Piperazine Ferulate on TGF-β_1-induced Renal Interstitial Fibroblast Activation

目的观察阿魏酸哌嗪对转化生长因子-β1(TGF-β1)诱导肾成纤维细胞表型活化的影响,探讨其抗肾间质纤维化的可能机制。方法体外培养正常大鼠肾脏成纤维细胞(NRK-49F),利用MTT观察阿魏酸哌嗪对TGF-β1诱导细胞活力的影响;免疫细胞化学法观察阿魏酸哌嗪对TGF-β1诱导细胞α-平滑肌肌动蛋白(α-SMA)、结缔组织生长因子(CTGF)蛋白表达的影响;实时荧光定量逆转录-聚合酶链反应(realtime RT-PCR)检测阿魏酸哌嗪对TGF-β1诱导细胞α-SMA及CTGFmRNA表达的作用,双抗体夹心ABC-ELISA法检测阿魏酸哌嗪对TGF-β1诱导细胞上清型胶原(Col)、纤维连接蛋白(FN)表达的影响。结果TGF-β1可以显著增加大鼠肾脏成纤维细胞活力,上调细胞α-SMA、CTGF蛋白和mRNA的表达以及Col、FN的表达。与TGF-β1刺激组相比,阿魏酸哌嗪能部分抑制TGF-β1诱导的细胞活力增加、α-SMA、CTGF的表达和细胞外基质(ECM)的合成。结论阿魏酸哌嗪可以在一定程度上拮抗TGF-β1诱导的肾纤维化效应。

Objective To investigate the effects of piperazine ferulate （PF） on TGF-β1-indueed renal interstitial fibroblast activation, and to explore the mechanism of PF in the prevention of renal tubulointerstitial fibrosis. Methods In cultured normal rat renal kidney fibroblast （NRK-49F）, the effect of PF on the TGF-β1- induced cell vitality was observed by MTT. The protein expression levels of a-smooth muscle actin （a-SMA）, connective tissue growth factor （CTGF） induced by TGF-β1 were evaluated by immunoeytochemistry. The levels of α-SMA, CTGF mRNA expression induced by TGF-β1 were determined by quantitative real time fluroscent polymerase chain reaction. The levels of collagen type I （Col I ） and fibroneetin（FN） protein expression were examined by Enzyme Linked Immunosorbent Assay （ELISA）. Results TGF-β1 may markedly increase the cell vitality, a-SMA and CTGF expression, FN and Col I protein expression（P〈0. 05）. Compared with TGF-β1- induced group, PF can partly decrease the cell vitality, level of a-SMA and CTGF expression and extraeellular matrix（ECM） synthesis induced by TGF-β1 （P〈0. 05）. Conclusion PF may inhibit TGF-β1-indueed fibrosis in the renal fibroblast to a certain extent.