高浓度尿酸对人近端肾小管上皮细胞氧化应激的影响

Effects of high concentrations of uric acid on oxidative stress of human renal tubular epithelial cells

目的观察高浓度尿酸对人近端肾小管上皮细胞氧化应激的影响,探讨其可能的作用途径。方法体外培养人近端肾小管上皮HK-2细胞,以720μmol/L的尿酸干预0、12、24、48 h后收集细胞;DCHF-DA染色检测细胞内活性氧(ROS)含量,分光光度计检测细胞上清液中的丙二醛(MDA)、总超氧化物岐化酶(T-SOD),MitoSOXTM染色观察活细胞线粒体中ROS的生成情况。结果尿酸干预12 h时细胞内总ROS含量较未干预时无明显变化,细胞上清中MDA含量、T-SOD活性亦较未干预时无明显差异(P均>0.05);干预24 h时总ROS含量有所升高,MDA含量上升,T-SOD活性降低,48 h时上述变化更为明显(P均<0.05);线粒体内ROS也在尿酸干预24 h时合成上调,48 h时升高更为显著(P<0.05)。结论高浓度尿酸可诱导HK-2细胞氧化应激反应,线粒体ROS合成增多可能是其作用基础。

Objective To observe the effects of high concentrations of uric acid（ UA） on oxidative stress of human proximal renal tubular epithelial cells,and to investigate the possible pathway. Methods HK-2 cells were cultured in vitro and exposed to 720 μmol /L UA for 0 h,12 h,24 h and 48 h. The reactive oxygen species（ ROS） production in cells was detected by DCHF-DA staining. The MDA content and T-SOD activity in the supernatants were measured by Spectrophotometer. The mitochondrial ROS production was detected by MitoSOXTM staining. Results There were no significant changes in ROS production,MDA content and T-SOD activity of HK-2 cells exposed to UA for 12 h（ all P〈 0. 05）. The cell ROS production and MDA content increased,while T-SOD activity decreased when HK-2 cells were exposed to UA for24 h. Changes were more significant at 48 h（ all P〈 0. 05）. The mitochondrial ROS production was increased in HK-2cells exposed to UA for 24 h,and it increased more when exposed to UA for 48 h（ P〈 0. 05）. Conclusion High concentrations？ of UA can induce the oxidative stress in HK-2 cells through the mechanism of increasing the mitochondrial ROS production.