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SS31对大鼠肾小管上皮细胞缺氧复氧损伤的影响作用及其机制

Pretreatment of SS31 prevents hypoxia/reoxygenation-induced apoptosis in renal tubular epithelial cells
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摘要 目的研究线粒体靶向性小分子多肽-SS31对肾小管上皮细胞缺氧复氧(HR)损伤的影响及其作用机制。方法以大鼠肾小管上皮细胞株NRK-52E细胞为研究对象,建立细胞HR模型,分为对照组、Ss31组、HR组、HR+Ss31组。四甲基偶氮唑盐实验行细胞损伤检测,annexinV/PI双染色法行细胞凋亡检测,荧光显微镜观察活性氧、线粒体膜电位的变化,蛋白质印迹法检测衔接蛋白p66Shc和磷酸化衔接蛋白p-p66Shc表达,比较组间结果差异。结果HR组细胞活性明显下降,为对照组的(52.2±6.0)%(P〈0.05);HR+SS31组细胞活性有所提高,呈SS31剂量依赖性。与对照组相比较,HR组细胞凋亡率增加,为(23.03±1.04)%,HR+SS31组(100/μmol/L)细胞凋亡率较HR组下降,为(10.37±0.80)%(P〈0.05)。HR+SS31组(100ptmol/L)细胞内活性氧的产生明显低于HR组;HR+SS31组(100μmot/L)线粒体膜电位下降较HR组有所改善。对照组细胞中p66Shc和p-p66Shc表达很低;在HR组细胞中,p66Shc和p-p66Shc表达都显著升高,分别为0.93±0.05和0.85±0.04;HR+SS31组(100μmol/L)两种蛋白的表达有所降低,分别为0.58±0.06和0.45±0.04(P〈0.05)。结论SS31可减轻大鼠肾小管上皮细胞HR损伤,其机制可能是通过p66Shc依赖的通路来抑制HR诱导的细胞凋亡和坏死,从而减少细胞损伤。 Objective To investigate whether the pretreatment of SS31 could alleviate hypoxia/ reoxygenation (H/R)-induced injury by inhibiting p66Shc. Method The cultured rat renal proximal tubular cell line NRK52E cells were exposed to 24-h hypoxia (5% C02, 1% 02, and 94% N2) followed by 6-h reoxygenation (5% C02, 21% 02, and 74% N2). SS31 was added to the culture medium 4 h prior to the treatment. Then the cell viability, apoptosis, ROS and MTP were determined. In addition, Western blotting was used to detect the expression of p66Shc and p-p66Shc. Result H/R induced apoptotic cell death, accompanied with activation of total and p-p66Shc in NRK52E cells. Total p66Shc and p-p66Shc were detected at low levels in control NRK52E cells, and their levels were dramatically increased in cells after H/R treatment. Pretreatment with 100 μmol/L SS31 significantly prevented cell death and attenuated total p66Shc and p-p66Shc levels after H/R. Conclusion This study revealed that SS31 pretreatment serves a protective role against H/R-induced apoptosis of human renal tubular epithelial cells by suppressing p66Shc.
出处 《中华器官移植杂志》 CAS CSCD 北大核心 2014年第4期232-236,共5页 Chinese Journal of Organ Transplantation
基金 国家自然科学基金青年项目(81000310)
关键词 缺氧 再灌注 肾小管 上皮细胞 体外研究 大鼠 Anoxia Reperfusion Kidney tubular Epithelial cells In vitro Rats
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