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目的探讨胰升血糖素样肽-1(GLP-1)对晚期氧化蛋白产物(AOPPs)诱导足细胞损伤的影响。方法以小鼠永生性足细胞系为研究对象,分为对照(Con)组、AOPPs组、GLP-1组、AOPPs+GIP-1组,CCK8试剂盒检测足细胞增殖活性,DHE荧光探针检测细胞内活性氧簇(ROS)水平,光泽精化学发光法测定还原型辅酶II(NADPH)氧化酶的活性,酶标仪法检测SOD活性、MDA及还原型谷胱甘肽(GSH)含量;Western blot检测足细胞特异性标志蛋白podocin、nephrin的表达情况。结果 GLP-1可呈浓度依赖性地拮抗AOPPs诱导的足细胞增殖活性降低,减少ROS产生[荧光强度为(48.93±2.99)],降低NADPH氧化酶活性(较AOPPs组下降40.1%);增加SOD酶活性和GSH含量,减少MDA产生,并增加nephrin、podocin蛋白表达(P<0.05,分别为AOPPs组的1.29倍、1.44倍)。结论 GLP-1能减轻AOPPs诱导的足细胞损伤,可能与抑制NADPH氧化酶介导的氧化应激有关。
Objective To investigate the effect of glucagon-like peptide-1 (GLP-1) on podocyte injury induced by advanced oxidized protein product (AOPPs). Methods The immortalized mouse podocytes were divided into control group, AOPPs group, GLP-1 group, AOPPs + GIP-1 group and CCK8 kit for detection of podocyte proliferative activity. DHE fluorescence probe The level of reactive oxygen species (ROS), the activity of NADPH oxidase by spectrophotometry and the activity of SOD and the content of glutathione (GSH) and glutathione (GSH) were detected by microplate reader. blot detection podocin, nephrin expression of podocyte specific marker protein. Results GLP-1 antagonized AOPPs-induced decrease of podocyte proliferative activity and ROS production in a concentration-dependent manner [fluorescence intensity (48.93 ± 2.99)], decreased NADPH oxidase activity (40.1% compared with AOPPs group) Enzyme activity and GSH content, decreased MDA production, and increased nephrin and podocin protein expression (P <0.05, respectively, 1.29 times AOPPs group, 1.44 times). Conclusion GLP-1 can reduce AOPPs-induced podocyte injury, which may be related to the inhibition of NADPH oxidase-mediated oxidative stress.