活性氧自由基在肾小管上皮细胞necroptosis中的作用

发布时间：2018-04-01 18:33

本文选题：活性氧自由基　切入点：necroptosis　出处：《肾脏病与透析肾移植杂志》2017年03期

【摘要】：目的:探讨活性氧自由基(ROS)在肾小管上皮细胞necroptosis中的作用。方法:构建肾小管上皮细胞HK-2细胞necroptosis模型,检测其ROS升高程度。并使用NADPH酶抑制剂Apocynin抑制HK-2细胞necroptosis模型中ROS的生成,通过流式细胞计数及检测necroptosis的关键蛋白观察HK-2细胞necroptosis的变化。结果:使用肿瘤坏死因子α、苄氧羰酰-缬氨酰-丙氨酰-天冬氨酰-氟甲基酮及抗霉素A成功建立了HK-2细胞necroptosis模型,并观察到HK-2细胞发生necroptosis时ROS显著升高(43.29±2.49 vs 25.90±1.27,P0.001),而使用necrostatin-1抑制necroptosis后ROS生成受到抑制(35.58±1.08 vs 43.29±2.49,P=0.002)。当对necroptosis模型使用Apocynin干预时,HK-2细胞ROS明显下降(30.71±2.82 vs 43.29±2.49,P0.001),并且流式细胞计数结果显示坏死细胞比例减少(2.00%±0.30%vs 6.99%±2.79%,P0.001),同时受体相关蛋白3和混合系列蛋白激酶样结构域的磷酸化水平降低。结论:ROS参与了HK-2细胞的necroptosis,并且通过抑制ROS的生成可减少necroptosis发生,提高损伤状态下HK-2细胞存活率,减轻急性肾小管坏死。
[Abstract]:Aim: to investigate the role of reactive oxygen species (Ros) in necroptosis of renal tubular epithelial cells. Methods: the necroptosis model of renal tubular epithelial cells (HK-2) was constructed, and the degree of ROS elevation was detected. Apocynin, a inhibitor of NADPH enzyme, was used to inhibit the production of ROS in necroptosis model of HK-2 cells. The changes of necroptosis in HK-2 cells were observed by flow cytometry and the key proteins of necroptosis. Results: tumor necrosis factor- 伪, benzoxycarbonyl-valeryl-alanine-aspartyl-fluoromethyl ketone and anti-mycin A were successfully established. The necroptosis model of HK-2 cells was established. It was also observed that ROS increased significantly during necroptosis in HK-2 cells (43.29 卤2.49 vs 25.90 卤1.27p 0.001g / L), while ROS production was inhibited by necrostatin-1 treatment with necroptosis 35.58 卤1.08 vs 43.29 卤2.49 P0. 002. The ROS of HK-2 cells decreased by 30.71 卤2.82 vs 43.29 卤2.49 P0.001 after Apocynin intervention in necroptosis model, and the results of flow cytometry were significant. The percentage of necrotic cells decreased by 2.00% 卤0.30%vs 6.99% 卤2.79% 卤2.79%, and the phosphorylation level of receptor-associated protein 3 and mixed series protein kinase-like domains decreased. Conclusion: Ros is involved in the necrotosis of HK-2 cells and can reduce the occurrence of necroptosis by inhibiting the production of ROS. The survival rate of HK-2 cells was increased and the acute tubular necrosis was alleviated.
【作者单位】：南方医科大学;广东省人民医院广东省医学科学院;中山大学中山医学院高血压研究所;
【基金】：国家临床重点专科建设项目国家自然科学基金面上项目(81570609) 广东省自然科学基金(2014A030313545)
【分类号】：R692

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