p38 MAPK信号通路介导晚期氧化蛋白产物诱导的肾小管上皮细胞间充质转分化  

P38 MAPK signaling pathway mediates advanced oxidation protein product- induced epithelial-to-mesenchymal transition in tubular cells

作　　者：黄丽丽[1,2] 祝小林 邓伟谦 段娜[2] 梁秀洁 王悦[2] 郭婷婷[2] 束双双[2] 向晓红[2] 姜婷婷[2] 汤珣[2] 章俊[2] HUANG Lili ZHU Xiaolin DENG Weiqian DUAN Na LIANG Xiujie WANG Yue GUO Tingting SHU Shuangshuang XIANG Xiaohong JIANG Tingting TANG Xun ZHANG Jun(Department of Nephrology, Fifth Affiliated Hospital, Southern Medical University, Guangzhou 510900, China Department of Nephrology, Zhujiang Hospital of Southern Medical University, Guangzhou 510280, China Department of Nephrology, Zhongshan Guzhen People＇s Hospital, Zhongshan 528421, China)

机构地区：[1]南方医科大学第五附属医院肾内科,广东广州510900 [2]南方医科大学珠江医院肾内科,广东广州510280 [3]中山市古镇人民医院肾内科,广东中山528421

出　　处：《南方医科大学学报》2016年第9期1209-1214,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81202842);广州市科技计划项目(11C22120703);广东省科技计划项目(2013B021800149);广东省自然科学基金(S2011010004053;10151051501000030;S2011040003566);中山市科技计划项目(2014A1FC085)~~

摘　　要：目的探讨p38丝裂原活化蛋白激酶(p38 MAPK)信号途径是否介导晚期氧化蛋白产物(AOPP)诱导肾小管上皮细胞间充质转分化(EMT)。方法用次氯酸氧化牛血清白蛋白(BSA)制备的AOPP-BSA刺激体外培养的人肾小管上皮细胞(HK-2细胞),用Western blotting检测细胞p38 MAPK和磷酸化p38 MAPK的表达;用p38 MAPK磷酸化抑制剂SB203580预处理细胞,并予AOPP-BSA刺激,用Western blotting和实时荧光定量PCR分别检测EMT标志性蛋白E-cadherin、vimentin和内质网应激标志性蛋白GRP78和m RNA表达;用内质网应激(ERS)阻断剂salubrinal预处理细胞,并予AOPP-BSA刺激,用Western blotting检测细胞p38 MAPK和磷酸化p38 MAPK的表达。结果 AOPP-BSA能使细胞p38 MAPK磷酸化;用SB203580抑制p38 MAPK磷酸化可显著抑制AOPP-BSA下调的E-cadherin和上调的vimentin和GRP78的表达;用salubrinal抑制ERS可有效地抑制AOPP-BSA诱导的p38 MAPK磷酸化。结论 p38 MAPK信号途径可能参与了AOPPs诱导肾小管上皮细胞发生EMT的过程,且p38 MAPK受ERS调控。Objective To investigate whether the p38 mitogen-activated protein kinase （MAPK） signaling pathway mediates advanced oxidation protein products （AOPPs）-induced epithelial-to-mesenchymal transition （EMT） in tubular cells. Methods Human proximal tubular cells （HK-2 cells） exposed to AOPP-bovine serum albumin （BSA） were examined for expressions of p38 MAPK and phosphorylated p38 MAPK using Western blotting. Western blotting and quantitative RT-PCR were used to examine the protein and mRNA expressions of EMT markers E-cadherin and vimentin and endoplasmic reticulum stress marker glucose-regulated protein （GRP） 78 in cells treated with SB203580 （an inhibitor of the p38 MAPK signaling pathway） prior to AOPP exposure. The cells treated with AOPPs following pretreatment with salubrinal （an inhibitor of endoplasmic reticulum stress） were also examined for expressions of p38 MAPK and phosphorylated p38 MAPK. Results AOPP treatment induced the phosphorylation of p38 MAPK in HK-2 cells. AOPP-induced decrease in E-cadherin expression and overexpression of vimentin and GRP78 were partly inhibited by pretreatment of the cells with SB203580. Salubrina partly suppressed AOPP-induced phosphorylation of p38 MAPK in the cells. Conclusion p38 MAPK signaling pathway, which is regulated by endoplasmic reticulum stress, might mediate AOPP-induced EMT in HK-2 cells.

关 键 词：P38 MAPK 间充质转分化 晚期氧化蛋白产物 内质网应激 HK-2细胞 

分 类 号：R692[医药卫生—泌尿科学]