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目的明确氧化低密度脂蛋白(ox-LDL)对血管内皮细胞中巨噬细胞移动抑制因子(MIF)表达的调控作用,检测MIF对血管内皮细胞增殖、迁移的可能调控作用。方法用不同剂量的ox-LDL处理人脐静脉血管内皮细胞(HUVECs),Western-blot检测MIF蛋白的表达。利用腺病毒介导MIF在HUVECs中的表达,分别通过Trans-well细胞穿膜实验和细胞划痕实验检测MIF对HUVECs迁移的调控作用。分别通过EdU染色、MTT细胞增殖检测和流式细胞术检测细胞周期,分析MIF对HUVECs增殖的影响。用荧光定量PCR检测MIF对HUVECs中MMP2和CXCR4表达的影响。结果Ox-LDL可特异调控HUVECs中MIF的表达。Trans-well细胞穿膜实验和细胞划痕实验表明,利用腺病毒介导MIF高表达可显著抑制HUVECs的迁移。EdU染色、MTT和细胞周期检测结果显示MIF对HUVECs增殖无明显作用。定量PCR结果显示过表达MIF可显著抑制HUVECs中CXCR4的表达(P<0.01)。结论 MIF通过降低CXCR4表达来抑制血管内皮细胞的迁移,但对血管内皮细胞的增殖没有影响。
Objective To investigate the regulatory effect of ox-LDL on the expression of macrophage migration inhibitory factor (MIF) in vascular endothelial cells and the possible regulation of MIF on the proliferation and migration of vascular endothelial cells. Methods Human umbilical vein endothelial cells (HUVECs) were treated with different doses of ox-LDL and the expression of MIF protein was detected by Western-blot. Adenovirus-mediated MIF expression in HUVECs was used to detect the regulatory effect of MIF on the migration of HUVECs by Trans-well cell transmembrane assay and cell scratch assay, respectively. The cell cycle was detected by EdU staining, MTT cell proliferation assay and flow cytometry, respectively. The effects of MIF on the proliferation of HUVECs were analyzed. The effect of MIF on the expression of MMP2 and CXCR4 in HUVECs was detected by real-time PCR. Results Ox-LDL could specifically regulate the expression of MIF in HUVECs. Trans-well cell transmembrane and cell scratch assays showed that adenovirus-mediated MIF overexpression significantly inhibited the migration of HUVECs. EdU staining, MTT and cell cycle test results showed that MIF had no significant effect on the proliferation of HUVECs. Quantitative PCR results showed that overexpression of MIF significantly inhibited CXCR4 expression in HUVECs (P <0.01). Conclusion MIF can inhibit the migration of vascular endothelial cells by decreasing the expression of CXCR4, but has no effect on the proliferation of vascular endothelial cells.