MG132对草酸钙肾结石模型TGF-β1及EMT的影响

Effects of MG132 on the TGF-β1 and EMT in the model of calcium oxalate stone

目的探讨泛素蛋白酶体抑制剂MG132对草酸钙肾结石模型转化生长因子β1(TGF-β1)/Smad信号通路及上皮间质转化(EMT)的影响。方法 MDCK培养后分为3组(空白对照组、草酸组、草酸+MG132组)分别予以相应干预。运用Western blot蛋白印迹分析检测上皮钙黏蛋白(E-Cadherin)、N-钙黏着蛋白(N-Cadherin)、兔抗人波形蛋白(Vimentin)、TGF-β1、Smad7的表达。采用细胞可渗透性荧光探针2′-,7′-二氯荧光素二乙酸酯(DCFH-DA)测量细胞活性氧(ROS)水平。结果草酸会使TGF-β1、N-Cadherin、Vimentin表达量增加,使Smad7、E-Cadherin表达下降。MG132会降低草酸引起的细胞ROS的增加。结论泛素蛋白酶体抑制剂MG132通过抑制TGF-β1/Smad通路、影响肾小管上皮细胞的转分化从而发挥抗肾间质纤维化作用。

Objective To investigate the effects of ubiquitin proteasome inhibitor MG132 on the transforming growth factor-βl （TGF-β1）and epithelial-mesenchymal transformation （EMT）in the model of calcium oxalate stone. Methods MD-CK cells were divided into 3 groups：control group,oxalate group, and MG132＋oxalate group. The protein expressions of E-cadherin,N-cadherin,vimentin,TGF-β1 and Smad7 were detected with Western blot. The level of reactive oxygen species （ROS） was measured with cell permeable fluorescence probe 2,7- dichlorodi-hydrofluorescein diacetate （DCFH-DA）. Results Ox- Mate increased the levels of TGF-β1, N-cadherin and vimentin while decreased the levels of Smad7 and E-cadherin. MG132 markedly reduced the ROS induced by oxalate. Conclusion MG132 affects the epithelial mesenchymal transformation of MD- CK cells and renal fibrosis by inhibiting the TGF-β1/Smads signaling pathway.