不同浓度AOPPs对肾小球足细胞MCP-1表达及合成的影响

The effect of different concentration of AOPPs on MCP-1 expression in glomerular podocyte

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 observe the different concentrations of advanced oxidative protein products（AOPPs） on monocyte chemotactic protein 1（MCP-1） expression in mouse podocyte clone 5（MPC5）, and explore the effect of AOPPs on the progression of diabetic nephropathy. Methods MPC5 was cultured in vitro, and then set up five groups ： normal control group, the MSA group, different concentrations of AOPPs intervention group（50,100,200 μmol / L）. Real-time fluorescent quantitative PCR was used to detect MCP-1 m RNA expression in cells; MCP-1 protein was detected by ELISA. Results Compared with control group and MSA group, MCP-1 protein was increased significantly after induced by AOPPs for 24 h（P〈0.01）, and increased gradually along with the higher concentration of AOPPs. Compared with the control group and MSA group, AOPPs induced MCP-1 m RNA expression obviously, and increased with the increase of concentration of AOPPs after 24 h（P 〈0.01）. Conclusion AOPPs could induced the expression of MCP-1 in mouse podocyte clone 5 in dose dependent manor. MCP-1 may be participated in the development of diabetic kidney disease.