U0126对TGF-β1诱导足细胞炎症因子分泌和miR-155表达的影响

Effect of U0126 on inflammatory cytokine secretion and miR-155 expression in podocyte induced by TGF-β1

目的 探讨细胞外调节蛋白激酶(erk)1/2通路抑制剂U0126对转化生长因子(transforming growth factor,TGF)-β1诱导足细胞炎症因子分泌和微RNA(microRNA,miR)-155表达的影响。方法 体外培养小鼠足细胞,随机分为正常对照组、TGF-β1组和TGF-β1+U0126组,使用Westernblot法检测erk1/2磷酸化水平,ELISA法检测足细胞上清TNF-α及IL-6蛋白表达水平,RT-qPCR法检测足细胞miR-155表达水平。结果 与正常对照组相比,TGF-β1诱导足细胞后,erk1/2磷酸化水平明显上升,足细胞上清IL-6和TNF-α蛋白分泌增多,足细胞miR-155表达显著上调,差异均有统计学意义(P<0.05)。与TGF-β1组相比,TGF-β1+U0126组erk1/2磷酸化水平被抑制,足细胞上清IL-6和TNF-α蛋白分泌减少,足细胞miR-155表达下降,差异均有统计学意义(P<0.05)。结论 erk1/2通路抑制剂U0126可抑制足细胞炎症因子IL-6、TNF-α分泌,下调足细胞miR-155的表达。

Objective To investigate the effects of U0126,an extracellular regulated protein kinases(erk)1/2 pathway inhibitor,on inflammatory cytokine secretion and (microRNA,miR)-155 expression in podocytes induced by (transforming growth factor,TGF)-β1. Methods Mouse podocytes were cultured in vitro and randomly divided into normal control group,TGF-β1 group and TGF-β1+U0126 group.The erk1/2 phosphorylation level was detected by Western blot,the expressions of supernatant TNF-α and IL-6 were detected by ELISA,and the miR-155 expression in podocyte was detected by RT-qPCR. Results Compared with the normal control group,the phosphorylation level of erk1/2 significantly increased,the secretion of supernatant IL-6 and TNF-α protein increased,and the expression of miR-155 in podocytes significantly up-regulated after TGF-β1 inducement,difference was statistically significant( P <0.05).Compared with the TGF-β1 group,the phosphorylation level of erk1/2 in TGF-β1+U0126 group was inhibited,the secretions of IL-6 and TNF-α protein in podocyte supernatant decreased,and the miR-155 expression in podocyte decreased,difference was statistically significant( P <0.05). Conclusion erk1/2 pathway inhibitor U0126 can inhibit the secretions of inflammatory factors such as IL-6 and TNF-α,and down-regulate miR-155 expression in podocyte.