过表达趋化因子受体4的内皮祖细胞影响血管平滑肌细胞迁移、增殖

目的 研究过表达趋化因子受体4(chemokine receptor 4,CXCR4)的内皮祖细胞(endothelial progenitor cells,EPCs)对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)迁移、增殖的影响.方法 分离培养并鉴定大鼠骨髓来源EPCs.CXCR4的重组腺病毒感染EPCs后,流式细胞术、实时荧光定量PCR(qRT-PCR)分别检测细胞膜CXCR4受体及mRNA表达,CCK-8法、Transwell法分别检测EPCs增殖活性、迁移能力.建立过表达CXCR4的EPCs与VSMCs非接触共培养模型,Transwell法、CCK-8法检测EPCs对VSMCs的迁移、细胞存活的影响.结果 CXCR4重组腺病毒感染EPCs 48 h后,EPCs细胞膜CXCR4受体和mRNA表达明显增高(P＜0.01),过表达CXCR4对EPCs增殖活性无明显影响,但可增强EPCs定向迁移能力(P＜0.01).共培养模型中,SDF-1α诱导下,CXCR4组VSMCs迁移、细胞存活率明显降低(P＜0.05),无SDF-1α诱导下空白对照组、GFP组及CXCR4组VSMCs的迁移、细胞存活率整体高于SDF-1α诱导下各组(P＜0.05).结论 过表达CXCR4可增强EPCs的迁移能力,在SDF-1/CXCR4调控轴中加强EPCs对VSMCs迁移和增殖的抑制作用,可用于防治血管再狭窄的细胞治疗.

Effect of CXCR4 over-expressed endothelial progenitor cells on migration and proliferation in vascular smooth muscle cells

Objective To determine the effects of endothelial progenitor cells (EPCs) with overexpression of chemokine receptor 4 (CXCR4) on the migration and proliferation of vascular smooth muscle cells (VSMCs).Methods EPCs were isolated from rat bone marrow,and cultivated and identified,and then were infected with recombinant adenovirus of CXCR4.Flow cytometry and qRT-PCR were used to detect the membrane expression of CXCR4 receptor and the mRNA expression of the molecule respectively.CCK-8 assay was applied to detect the proliferation of EPCs,and Transwell assay was carried out to evaluate the migration of EPCs.Transwell culture plate was constructed into a non-contact co-cuhure model between CXCR4 over-expressed EPCs and VSMCs.EPCs' effects on the migration of VSMCs and cell survival were detected with Transwell chamber test and CCK-8 assay.Results After the EPCs were infected with recombinant adenovirus of CXCR4 for 48 h,the expression of membrane CXCR4 receptor and CXCR4 mRNA was significantly increased (P ＜ 0.01),and the over-expression of CXCR4 had no significant effect on the proliferative activity of EPCs,but enhanced the oriented migration ability of EPCs (P ＜0.01).In the coculture model,the migration and cell viability of VSMCs were obviously lower after the induction of SDF-1 a (P ＜ 0.05).In absence of SDF-1 a induction,the migration and cell viability were obviously higher in the blank control cells,GFP infected cells and CXCR4 over-expressed cells than the corresponding cells after induction of SDF-1 a (P ＜ 0.05).Conclusion The over-expression of CXCR4 could increase the migration ability of EPCs,and enhance its inhibitory effects on the migration and proliferation of VSMCs through SDF-1/CXCR4 axis.So it can be used for cell therapy in the prevention and control of vascular restenosis.