GLO Ⅰ敲减对AGEs诱导人肾小球系膜细胞氧化应激的影响  

Effect of knockdown of GLO Ⅰ on AGEs-induced oxidative stress of human renal mesangial cells

作　　者：徐莹 李春 操轩[1] XU Ying;LI Chun;CAO Xuan(Department of Nephrology,Affiliated Hospital/Huangshi Municipal Central Hospital, Hubei Institute of Science and Technology,Huangshi,Hubei 435000,China)

机构地区：[1]湖北理工学院附属医院/湖北省黄石市中心医院肾内科

出　　处：《重庆医学》2019年第20期3427-3430,共4页Chongqing medicine

基　　金：湖北省自然科学基金项目(2013CFC058)

摘　　要：目的分析乙二醛酶Ⅰ(GLOⅠ)敲减对晚期糖基化终末产物(AGEs)诱导人肾小球系膜细胞(HRMCs)氧化应激的影响,并探讨其可能机制。方法利用酶联免疫吸附试验(ELISA)分析糖尿病肾病(DN)患者尿液中GLOⅠ的水平;小干扰RNA(siRNA)对HRMCs中GLOⅠ进行敲减,Western blot验证敲减效果;H2DCFDA荧光探针标记法检测细胞内ROS活性;Q-PCR和Western blot检测P22phox表达;Western blot检测PI3K/AKT信号蛋白和p38信号蛋白的活化。结果与对照组(健康个体)相比,GLOⅠ在DN患者尿液中水平降低;GLOⅠ敲减可导致AGEs诱导的细胞ROS活性进一步增加,P22phox基因和蛋白表达进一步增加,以及AKT和p38的磷酸化进一步增加。结论GLOⅠ敲减可使AGEs诱导的HRMCs氧化应激进一步增强,这可能与进一步上调PI3K/AKT和p38信号通路活性有关。Objective To analyze the effect of glyoxalase Ⅰ(GLOⅠ) knockdown on AGEs-induced oxidative stress of human renal mesangial cells,and to probe its possible mechanism.Methods The urine GLOⅠlevel in the patients with diabetes nephrosis (DN) was detected by ELISA.The knockdown of GLOⅠ in HRMCs was conducted by RNA interference (RNAi) and its effect was validated by Western blot.The intracellular ROS activity was analyzed by H2DCFDA fluorescence probe labeling assay.The expression of P22phox was determined by Q-PCR and Western blot.The activation of PI3K/AKT signal and p38 signal was analyzed by Western blot.Results Compared with the control group,the level of urine GLOⅠ in the DN patients was decreased.GLOⅠ knockdown further increased AGEs-induced cellular ROS activity of HRMCs, further enhanced AGEs-induced P22phox gene and protein expression,and further increased the phosphorylation of AKT and p38.Conclusion GLOⅠ Knockdown of GLOⅠcan increase AGEs-induced oxidative stress of HRMCs,which is possibly related with further activating PI3K/AKT and p38 signal activity.

关 键 词：乙二醛酶Ⅰ 糖尿病肾病 氧化应激 

分 类 号：R587.1[医药卫生—内分泌]