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目的观察不同浓度晚期蛋白氧化产物(AOPPs)对体外培养足细胞(MPC5)内单核细胞趋化蛋白-1(MCP-1)表达的影响,探讨AOPPs对糖尿病肾病病情进展的影响。方法以体外培养的足细胞进行研究,共设立5个组,包括正常对照组、小鼠血清白蛋白(MSA)组、不同浓度的AOPPs(50、100、200μmol/L)干预组。实时荧光定量PCR检测足细胞内MCP-1 m RNA的表达,用酶联免疫(ELISA)法检测培养足细胞上清中MCP-1蛋白的表达。结果与对照组及MSA组相比,AOPPs诱导足细胞24 h后,上清MCP-1蛋白浓度明显升高(P<0.01),并且随AOPPs浓度的增大而逐渐增高,呈浓度依赖性升高;与对照组及MSA组相比,AOPPs诱导足细胞24 h后,细胞内MCP-1m RNA的表达明显增强(P<0.01),并且随浓度的增大而逐渐增强。结论 AOPPs可诱导肾小球足细胞MCP-1表达升高,并且具有浓度依赖性,升高的MCP-1可能参与了糖尿病肾病病情的发展。
Objective To investigate the effects of different concentrations of advanced protein oxidase (AOPPs) on the expression of monocyte chemoattractant protein-1 (MCP-1) in cultured podocytes (MPC5) in vitro and the effect of AOPPs on the progression of diabetic nephropathy. Methods Five podocytes were cultured in vitro, including normal control group, mouse serum albumin (MSA) group, and different concentration of AOPPs (50,100,200μmol / L). The expression of MCP-1 mRNA in podocytes was detected by real-time fluorescence quantitative PCR. The MCP-1 protein expression in podocytes was detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with control group and MSA group, the concentration of MCP-1 protein in supernatant of AOPPs induced by AOPPs increased significantly (P <0.01) and increased gradually with the increase of AOPPs concentration in a dose-dependent manner Compared with the control group and MSA group, the expression of MCP-1mRNA in podocytes induced by AOPPs increased significantly (P <0.01) 24 h after AOPPs treatment, and gradually increased with the increase of concentration. Conclusion AOPPs can induce MCP-1 expression in glomerular podocytes in a concentration-dependent manner. Increased MCP-1 may be involved in the development of diabetic nephropathy.