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目的观察髓过氧化物酶(MPO)对内皮细胞单核细胞趋化蛋白-1(MCP-1)表达的影响及二甲双胍的干预作用。方法应用western-blot检测内皮细胞内MCP-1、腺苷酸活化蛋白激酶α亚基(AMPKα)蛋白表达,酶联免疫吸附法检测细胞培养液中单核细胞趋化蛋白-1的含量变化。同时应用RNA干扰技术,进一步观察AMPKα的作用。结果MPO显著增加MCP-1的蛋白表达和培养液中MCP-1的浓度(P<0.01),随作用浓度和时间增加而显著增强。二甲双胍显著减少内皮细胞MCP-1的蛋白表达和培养液中MCP-1的浓度(P<0.01),而AMPKαSiRNA干预后,二甲双胍对内皮细胞MCP-1的影响作用显著减弱(P<0.01)。结论MPO可刺激内皮细胞MCP-1的蛋白表达和分泌,二甲双胍通过促使AMPKα磷酸化,激活AMPKα,抑制MPO对内皮细胞的促炎性反应,从而发挥其有益作用。
Objective To investigate the effect of myeloperoxidase (MPO) on the expression of monocyte chemotactic protein-1 (MCP-1) in endothelial cells and the effect of metformin on it. Methods The expression of MCP-1 and AMPKα in endothelial cells was detected by western-blot. The changes of MCP-1 in cell culture medium were detected by enzyme-linked immunosorbent assay. At the same time, RNA interference technology was used to further observe the effect of AMPKα. Results MPO significantly increased the protein expression of MCP-1 and the concentration of MCP-1 in culture medium (P <0.01), which was significantly increased with the increase of concentration and time. Metformin significantly reduced the MCP-1 protein expression in endothelial cells and MCP-1 concentration in culture medium (P <0.01). The effect of metformin on the expression of MCP-1 in endothelial cells was significantly attenuated by AMPKαSiRNA treatment (P <0.01). Conclusion MPO can stimulate the expression and secretion of MCP-1 in endothelial cells. Metformin can exert its beneficial effects by phosphorylating AMPKα, activating AMPKα and inhibiting the pro-inflammatory response of MPO to endothelial cells.