高糖通过激活人脐静脉内皮细胞ⅠκB-α/NF-κB途径诱导MCP-1的表达

High glucose induces monocyte chemoattractant protein-1 expression via stimulating NF-κB activity mediated by ⅠκB-α phosphorylation in the cultured human umbilical vein endothelial cells

目的观察高糖诱导人脐静脉内皮细胞NF-κB的活性和单核细胞趋化蛋白1(MCP-1)的表达,探讨糖尿病并发动脉粥样硬化的发病机制。方法在培养的人脐静脉内皮细胞中加入不同浓度葡萄糖,检测内皮细胞中MCP-1mRNA、MCP-1蛋白的表达,NF-κB的活性及IκB-α的磷酸化水平。结果高糖明显地诱导了血管内皮细胞NF-κB的活性、MCP-1的表达及IκB-α的磷酸化;NF-κB活性抑制剂明显地降低了高糖所诱导的MCP-1的表达。结论高糖通过诱导血管内皮细胞IκB-α的磷酸化激活NF-κB,从而诱导了MCP-1的表达。提示在糖尿病并发动脉粥样硬化的发病过程中,高血糖通过激活血管内皮细胞IκB-α/NF-κB途径诱导MCP-1的表达,进而发挥了重要的作用。

Objective To understand the mechanism for diabetes-induced atherosclerosis by observing monocyte chemoattractant protein-1 （MCP-1） expression and NF-κB activity in cultured endothelial cells with high concentration of glucose. Methods Human umbilical vein endothelial cells were isolated and cultured in M199 medium. RT-PCR was conducted to detect MCP-1 mRNA expression and ELISA was used to detect MCP-1 protein expression. Electrophoretic mobility shift assay （EMSA） was performed for NF-κB activity and Western blot analysis was carried out to detect phosphorylation of ⅠκB-α. Results High concentration of glucose significantly stimulated NF-κB activity and MCP-1 expression in the cultured cells, and the NF-κB specific inhibitor peptide （NF-κB SN50） markedly blocked the high glucose-induced MCP-1 expression and phosphorylation of ⅠκB-α. Conclusions High concentration of glucose induces MCP-1 expression via stimulating NF-κB activity mediated by phosphorylation of ⅠκB-α in vascular endothelial cells.