AGEs诱导培养人肾小球系膜细胞中NOX4及O_2~-水平

目的:观察晚期糖基化终末产物(AGEs)培养下人肾小球系膜细胞(HMCs)中还原型尼克安腺嘌吟二核苷酸氧化酶4(NOX4)蛋白及超氧阴离子(O-2)水平变化。方法:将正常浓度葡萄糖(5.6mmol/L)培养的HMCs经过无血清培养液同步化24h后分为两组:正常对照组5.6mmol/L葡萄糖+200mg/L牛血清白蛋白(BSA)]及AGEs组(5.6mmol/L葡萄糖+200mg/LAGEs),分别干预48h后用Western blotting检测NOX4蛋白表达,用Dihydroethidium(DHE)法检测O-2的荧光水平。结果:与正常对照组比较,AGEs组NOX4蛋白表达明显上调(P<0.01),O-2水平亦明显增加(P<0.01),两者呈显著正相关。结论:AGEs能激活HMCs中NOX4,引起氧化应激,可能与糖尿病肾病(DN)的发生有关。

The Level of NOX4 and O2- in Human Glomerular Mesangial Cells Cultured by AGEs

Objective: To investigate the change of NADPH oxidase subunit NOX4 and O2- in human glomerular mesangial cells(HMCs) cultured by AGEs. Method: After normal glucose(5.6mmol/L glucose) cultured human mesangial cells were synchronized 24 hours, HMCs were divided into two groups: the control group(5.6 mmol/L glucose+200mg/L BSA) and the AGEs group(200mg/L AGEs) for 48 hours.NOX4 protein were detected by Western blotting,fluorescence of O2- were assessed by dihydroethidium(DHE). Results: Compared with the control group,the protein of NOX4 significantly increased in HMCs cultured by AGEs(P<0.01) and the fluorescence of O2- also raised(P<0.01). Conclusion: AGEs can cause the increasing of NOX4 in cultured HMCs to produce O2-,which induced oxidative stress. Maybe,it is related with the occurrence of diabetic nephropathy.