PLK1靶向β-catenin介导慢性肾脏病炎症状态下足细胞转分化  

PLK1 mediates epithelial-mesenchymal transition of podocytes under inflammatory condition of chronic kidney disease via β-catenin

作　　者：王子阳 范明华[2] 吕智美[1] 王荣[1] WANG Ziyang;FAN Minghua;LYU Zhimei;WANG Rong(Department of Nephrology,Shandong Provincial Hospital Affiliated to Shandong University,Jinan 250021,China;Department of Obstetrics and Gynecology,Second Hospital of Shandong University,Jinan 250033,China)

机构地区：[1]山东大学附属省立医院肾内科,济南250021 [2]山东大学第二医院妇产科,济南250033

出　　处：《免疫学杂志》2018年第11期946-951,共6页Immunological Journal

基　　金：国家自然科学基金(81370834;81400732);山东省自然科学基金(ZR2016HP21)

摘　　要：目的探讨慢性肾脏病微炎症状态下PLK1(polo-like kinase 1)在足细胞上皮-间充质转分化(epithelial-mesenchymal transition,EMT)中的作用机制。方法以条件永生化小鼠足细胞系为研究对象进行分组:正常对照组;脂多糖(lipopolysaccharide,LPS)诱导组;PLK1 shRNA组(PLK1 shRNA+LPS)及Scramble shRNA组(Scramble shRNA+LPS)。利用Real-time PCR、Western blot检测足细胞中PLK1、结蛋白(desmin)mRNA及蛋白表达水平;Western blot检测PLK1对β-链蛋白(β-catenin)的作用;Western blot检测EMT相关分子E-钙粘素(E-cadherin)及波形蛋白(vimentin)的表达;Western blot检测糖原合成酶激酶3β(GSK-3β)的水平;Transwell迁移实验检测足细胞迁移能力。结果 LPS诱导炎症状态下,足细胞PLK1 m RNA及蛋白表达升高伴随β-catenin上调,同时,GSK-3β表达降低,vimentin呈现高表达,而E-cadherin呈现低表达;PLK1 shRNA转染足细胞可特异性敲低PLK1表达,下调β-catenin,伴随GSK-3β表达增加,逆转足细胞EMT的发生;且炎症状态下,足细胞Desmin表达增加;Transwell显示足细胞迁移能力增强。结论 LPS刺激可导致足细胞PLK1高表达,其可能通过负向调控β-catenin降解复合物(axin/GSK-3/APC)从而稳定β-catenin的表达,继而下调Vimentin,导致足细胞损伤及EMT的发生。To investigate the role of PLK1 in epithelial-mesenchymal transition（EMT） of conditionally immortalized mouse podocytes（MPC） under inflammatory condition of chronic kidney disease（CKD）, MPC were divided into control group, LPS induced group, PLK1 shRNA＋LPS group and Scramble shRNA＋LPS group. The expression of PLK1 and desmin were detected by Real-time PCR and Western blotting; the effects of PLK1 on β-catenin was measured by Western blotting; EMT associated proteins E-cadherin and vimentin were detected by Western blotting; the level of GSK-3β, which was participated in the degradation of β-catenin, were detected by Western blotting in different conditions; the motility of cultured podocytes in different conditions were tested by the transwell migration assay. Data showed that PLK1 was elevated in inflammatory condition induced by LPS, in paralled with the over expression of β-catenin and low expression of GSK-3β; mesenchymal marker vimentin was increased, whereas the expression of E-cadherin was decreased; the motility of cultured podocytes in inflammatory condition induced by LPS was enhanced compared with the control group; however, podocytes EMT can be reversed by PLK1 knockdown via transfection of PLK1 shRNA, accompanied by the decline of β-catenin and the raise of GSK-3β; what＇s more, podocytes injury were exhibited by increasing of desmin in inflammatory condition induced by LPS. Thus LPS could stimulates the expression of PLK1 in podocytes, at the same time PLK1 could stabilize β-catenin by regulating axin/GSK-3β/APC negatively, causing the down-expression of mesenchymal marker Vimentin, resulting in the lesion and EMT of podocytes ultimately.

关 键 词：脂多糖 足细胞 EMT PLK1 Β-CATENIN 

分 类 号：R392.12[医药卫生—免疫学]