南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (05): 659-.

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晚期蛋白氧化产物通过氧化应激诱导肾小管上皮细胞转分化

章俊,邱敏姿,马亚琼,卜阳,杨蕾,汤珣   

  • 出版日期:2014-05-20 发布日期:2014-05-20

Advanced oxidation protein products induce epithelial-to-mesenchymal transition in
cultured human proximal tubular epithelial cells via oxidative stress

  • Online:2014-05-20 Published:2014-05-20

摘要: 目的探讨晚期蛋白氧化产物(AOPP)对人近端肾小管上皮细胞转分化(EMT)的影响及作用机制。方法实验共3部分:⑴
分别用不同浓度AOPP及BSA刺激细胞24 h后检测α-SMA、E-Cadherin蛋白表达;⑵AOPP刺激细胞不同时间后检测α-SMA、
E-Cadherin表达;⑶分别予BSA 、AOPP以及用DPI、C-SOD预处理后再加入AOPP刺激细胞,检测α-SMA、E-Cadherin表达和不
同时间的MDA 含量及SOD、CAT、GSH-px 活力。结果AOPP 以浓度和时间依赖方式诱导HK-2 细胞α-SMA 表达上调、
E-Cadherin表达下调。AOPP引起细胞MDA含量上升,SOD、CAT和GSH-px活力下降。DPI、C-SOD预处理细胞可以部分抑
制AOPP诱导的α-SMA表达上调和E-Cadherin表达下调;降低MDA含量,提高SOD、CAT和GSH-px活力。结论AOPP通过
氧化应激诱导肾小管上皮细胞EMT,抑制NADPH氧化酶活性及抗氧化治疗可以抑制肾小管上皮细胞EMT,延缓肾间质纤维
化的进展。

Abstract: Objective To investigate the effects of advanced oxidation protein products (AOPP) on epithelial-to-mesenchymal
transition (EMT) in cultured human proximal tubular epithelial cells (HK-2) and explore the mechanism. Methods HK-2 cells
treated with 50, 100, 200, and 400 μg/ml AOPP or 50 μg/m bovine serum albumin (BSA) for 24 h, or with 200 μg/ml AOPP for
0.5, 1, 3, 6, 12, and 24 h were examined for the protein expression of α-SMA and E-cadherin. In cells pretreated with
diphenyleneiodonium (DPI) or cytoplasmic superoxide dismutase (C-SOD), the effects of 50 μg/ml BSA and 200 μg/ml AOPP
were assessed on the expressions of α-SMA and E-cadherin, malondialdehyde (MDA) level, superoxide dismutase (SOD)
activity, catalase (CAT) activity, and glutathione peroxidase (GSH-px) activity. Results AOPP treatment up-regulated α-SMA
expression and down-regulated E-cadherin expression in a dose- and time-dependent fashion. AOPP exposure of the cells
resulted in increased MDA level and lowered activities of SOD, CAT and GSH-PX. DPI and C-SOD partially attenuated the
effects of AOPP on α-SMA, E-cadherin, MDA, SOD, CAT and GSH-px. Conclusion AOPP can induce EMT in cultured HK-2
cells via oxidative stress, and this effect can be attenuated by inhibiting the activation of NADPH oxidase and using
antioxidants to delay the progression of renal interstitial fibrosis.