尿酸对人肾小管上皮细胞线粒体氧化损伤与凋亡的影响  

Effects of uric acid on mitochondrial oxidative damage and apoptosis in human renal tubularepithelial cells

作　　者：张涛[1] 李英[1] 迟雁青[1] 任韫卓[2] 刘茂东[1] 牛洪琳[1] 

机构地区：[1]河北医科大学第三医院肾内科,石家庄050081 [2]河北医科大学病理学教研室

出　　处：《中华肾脏病杂志》2014年第5期356-362,共7页Chinese Journal of Nephrology

基　　金：河北省卫生厅科研基金项目（20110111）

摘　　要：目的观察尿酸对人肾小管上皮细胞线粒体氧化损伤及相关凋亡途径的影响，探讨其可能作用机制。方法体外培养人肾小管上皮细胞（HK-2），分别加入不同浓度的尿酸（480μmol/L、720μmol/L）进行干预，于0h、24h、48h收集细胞；应用MitoSOX染色检测活细胞线粒体中ROS的产生；JC-1染色检测线粒体膜电位的变化；Western印迹和免疫荧光化学法对凋亡诱导因子（AIF）和抗增殖因子（prohibitin）蛋白表达进行细胞亚定位及半定量分析；AnnexinV-FITC及PI双染法检测细胞凋亡。结果480μmol/L尿酸刺激细胞24h，线粒体ROS产量较正常对照组升高（P＜0．05），且随尿酸刺激浓度升高和时间延长逐渐上调，线粒体膜电位则逐渐降低。480μmol/L尿酸刺激细胞24h，细胞凋亡率和AIF蛋白表达均较正常对照组无明显变化（P，0．05），而刺激48h时两者升高且其差异有统计学意义（Pc0．05）；720μmol/L尿酸干预24h时，AIF蛋白表达和细胞凋亡率已较正常对照组升高（Pc0．05），干预48h时升高更为显著。480μmol/L尿酸刺激细胞24h，prohibitin蛋白表达较正常对照组下降（P〈0．05），720μmol/L尿酸干预48h，降低最为明显。结论尿酸可诱导HK-2细胞线粒体ROS合成增多，膜电位下降，线粒体相关蛋白prohibitin表达下调，激活线粒体途径导致细胞凋亡。Objective To observe the effects of uric acid （UA） on mitochondrial oxidative damage and apoptosis in renal tubular epithelial cells （HK-2）, and investigate the possible mechanism. Methods HK-2 cells were exposed to UA （480 μmol/L, 720 μmol/L） for different time （0 h, 24 h, 48 h） in vitro. The mitochondrial ROS production was detected by MitoSOX staining. The mitochondrial membrane potential was measured by JC - 1 staining. The expressions of prohibitin and AIF were examined by Western blotting and immunofluorescence cytochemistry. The cell apoptosis was measured by Annexin V-FITC/PI staining. Results The mitochondrial ROS production in HK-2 cells exposed to 480 p, mol/L UA was increased than that of control group at 24 h （P 〈 0.05）, and increased gradually with UA concentration and incubation time increasing, while the mitochondrial membrane potential was reduced at the same time. There were no significant changes in AIF expression and apoptosis rate of HK -2 cells exposed to 480 μmol/L UA for 24 h compared with that of control group （P 〉 0.05）, while both of them were up-regulated when HK-2 cells were exposed to 480 μmol/L UA for 48 h and 720 Ixmol/L UA for 24 h and 48 h （P 〈 0.05）. The prohibitin expression in HK-2 cells exposed to 480μmol/L UAwas reduced than that of control group at 24 h （P 〈 0.05）, and down- regulated gradually with UA concentration and incubation time increasing. Conclusion Uric acid can induce the mitochondrial ROS production increased, the mitochondrial membrane potential reduced, the prohibitin expression down-regulated and the mitochondrial apoptosis pathway activated in HK-2 cells.

关 键 词：尿酸 细胞凋亡 线粒体 肾小管上皮细胞 抗增殖因子 

分 类 号：R589.2[医药卫生—内分泌] R692[医药卫生—内科学]