摘要
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 effect of macrophage migration inhibitory factor（MIF）on the proliferation and migration of endothelial cells（HUVECs）. Methods MIF protein expression was determined by Western-blot assay using ox-LDL- treated human umbilical vein endothelial cells （HUVECs）. Enforced expression of MIF in HUVECs was mediated by recombinant MIF adenovirus.Trans-well migration assay and wound healing assay were performed to study the effect of MIF on HUVECs migration.Edu staining, MTI＇ assay and flow cytometry for cell cycle assay were performed to investigate the effect of MIF on HUVECs proliferation. Modulation of MMP2 and CXCR4 mRNA by MIF was detected by quantitative real-time PCR assay. Results Ox-LDL was found toregulate MIF expression in a dose-dependent＇ manner. The results of trans-well cell migration and wound healing assay showed that enforced expression of MIF could efficiently inhibit HUVECs migration. However, the results of Edu staining, MTr assay and flow cytometry indicated that MIF had no effect on HUVECs growth. Quantitative real-time PCR assay revealed that expression of CXCR4 mRNA, but not MMP2 mRNA,was significantly inhibited by MIF in HUVECs （P〈0.01）. Conclusion MIF inhibits endothelial cell migration through suppressing CXCR4 expression, with no effect on the endothelial cell proliferation.
出处
《热带医学杂志》
CAS
2013年第12期1441-1444,1447,共5页
Journal of Tropical Medicine
基金
国家自然科学基金(81070102
81270222)
广东省自然科学基金(S2011020005911
S2012010009453)
