南方医科大学学报

南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (09): 1209-.

• • 上一篇    下一篇

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

黄丽丽，祝小林，邓伟谦，段娜，梁秀洁，王悦，郭婷婷，束双双，向晓红，姜婷婷，汤珣，章俊   

  • 出版日期:2016-09-20 发布日期:2016-09-20

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

  • Online:2016-09-20 Published:2016-09-20

HTML

PDF

1115

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mRNA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。

Abstract: 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.