PPARγ通过PTEN调控AKT/FAK通路影响高糖条件下肾小管上皮细胞转分化  

PPARγaffects epithelial-mesenchymal transition in renal tubular epithe⁃lial cells under high glucose by regulating AKT/FAK signaling pathway through PTEN

作　　者：严瑞 杨雨星 刘玲伶[3] 王圆圆[3] 陈烨 郭兵[3] YAN Rui;YANG Yu-xing;LIU Ling-ling;WANG Yuan-yuan;CHEN Ye;GUO Bing(Department of Nephrology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Endocrinology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Guizhou Provincial Key Laboratory of Pathogenesis and Drug Research on Common Chronic Diseases,Gui-zhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学附属医院肾内科,贵州贵阳550004 [2]贵州医科大学附属医院内分泌与代谢病科,贵州贵阳550004 [3]贵州医科大学贵州省常见慢性疾病发病机制及药物研究重点实验室,贵州贵阳550025

出　　处：《中国病理生理杂志》2020年第9期1608-1615,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81960140);贵州省科技厅联合基金资助项目(黔科合LH字[2017]7183号)。

摘　　要：目的:探讨在高糖培养的肾小管上皮细胞中过氧化物酶体增殖物激活受体γ(PPARγ)对PTEN/AKT/FAK通路的调控机制及对肾小管上皮细胞上皮-间充质转化(EMT)的影响。方法:以高糖培养的肾小管上皮细胞NRK52E为研究对象,采用过表达及shRNA干扰技术分别上调和下调肾小管上皮细胞中PPARγ的表达,并培养48 h,应用细胞免疫荧光、Western blot和RT-qPCR方法检测PTEN及EMT相关蛋白的表达变化;然后,在过表达及敲减PTEN的情况下,分别给予PPARγ抑制剂GW9662及激动剂罗格列酮干预,进一步观察PPARγ对AKT/FAK通路的调节是否通过PTEN实现的。结果:过表达PPARγ组PPARγ和PTEN的mRNA及蛋白表达增加,E-cad⁃herin的表达明显上调,而vimentin和α-SMA蛋白表达显著下调(P<0.05);PPARγ敲减组PPARγ及PTEN的mRNA及蛋白表达水平明显降低,E-cadherin的表达也明显下调,而vimentin和α-SMA蛋白表达显著升高(P<0.05)。GW9662干预组PTEN的表达减少,而p-AKT(Thr308)、FAK及p-FAK(Tyr397)蛋白水平均明显升高(P<0.05);与GW9662组相比,GW9662+过表达PTEN组PTEN表达明显增加,并伴随p-AKT(Thr308)、FAK及p-FAK(Tyr397)蛋白水平的降低(P<0.05);罗格列酮干预组PTEN的表达增多,而p-AKT(Thr308)、FAK和p-FAK(Tyr397)蛋白水平降低(P<0.05);与罗格列酮组相比,罗格列酮+PTEN shRNA组PTEN的表达减少,相应的p-AKT(Thr308)、FAK和p-FAK(Tyr397)蛋白水平升高(P<0.05)。结论:PPARγ能调节肾小管上皮细胞PTEN的mRNA及蛋白表达水平,并影响肾小管上皮细胞的EMT;PPARγ对AKT及FAK通路的调节及对细胞EMT的影响主要是通过PTEN实现的。AIM:To investigate the role of peroxisome proliferator-activited receptor γ(PPARγ)in the regulation of PTEN/AKT/FAK signaling pathway and epithelial-mesenchymal transition(EMT)in renal tubular epithelial cells grown in high-glucose environment.METHODS:Renal tubular epithelial cells(NRK52E cells)cultured in high glucose were used as an in vitro model system.PPARγ was over-expressed or knocked down in these cells,and its effect on PTEN expression was determined by RT-qPCR,immunofluorescence and Western blot.The changes of EMT-related proteins were also measured.The PPARγ inhibitor GW9662 and the PPARγagonist rosiglitazone were used along with PTEN over-expression or knockdown to determine whether the effects of PPARγwere mediated through PTEN.RESULTS:PPARγover-expression resulted in the increased expression of PTEN at mRNA and protein levels,the up-regulation of E-cadherin,and the down-regulation of vimentin and α-SMA.Knockdown of PPARγexpression reduced the mRNA and protein levels of PTEN,down-regulated E-cadherin,and up-regulated vimentin andα-SMA(P<0.05).Treatment of the NRK-52E cells with GW9662 decreased PTEN expression and increased the protein levels of p-AKT(Thr308),FAK and p-FAK(Tyr397).These effects were rescued by PTEN over-expression.Treatment of the NRK-52E cells with rosiglitazone increased PTEN expression and decreased the protein levels of p-AKT(Thr308),FAK and p-FAK(Tyr397).These effects were rescued by PTEN knockdown.These changes were all statistically significant(P<0.05).CONCLUSION:PPARγ regulates the mRNA and protein expression of PTEN in renal tubular epithelial NRK52E cells,and affects EMT in renal tubular epithelial cells.The regulation of AKT/FAK signaling pathway by PPARγ is primarily mediated by PTEN.

关 键 词：高糖 过氧化物酶体增殖物激活受体Γ PTEN蛋白 肾小管上皮细胞 上皮-间充质转化 

分 类 号：R587.2[医药卫生—内分泌] R363.2[医药卫生—内科学]