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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。
Objective To investigate the effects of endothelial progenitor cells (EPCs) overexpressing chemokine receptor 4 (CXCR4) on the migration and proliferation of vascular smooth muscle cells (VSMCs). Methods The bone marrow derived EPCs were isolated and identified. CXCR4 recombinant adenovirus infected EPCs, flow cytometry, real-time quantitative PCR (qRT-PCR) were detected cell membrane CXCR4 receptor and mRNA expression, CCK-8 method, Transwell method were detected EPCs proliferation activity and migration ability. A non-contact co-culture model of EPCs overexpressing CXCR4 with VSMCs was established. Transwell assay and CCK-8 assay were used to detect the effect of EPCs on VSMCs migration and cell survival. Results CXCR4 recombinant adenovirus infected EPCs significantly increased the expression of CXCR4 receptor and mRNA (P <0.01) at 48 h. CXCR4 overexpression did not significantly affect the proliferation of EPCs but enhanced the ability of directional migration of EPCs (P <0.01) . In the co-culture model, the migration of VSMCs in CXCR4 group was significantly decreased after SDF-1α induction (P <0.05), and the migration of VSMCs in blank control group, GFP group and CXCR4 group without SDF- Higher than SDF-1αinduced in each group (P <0.05). Conclusion Overexpression of CXCR4 can enhance the migration of EPCs and enhance the inhibitory effect of EPCs on the migration and proliferation of VSMCs in the regulation axis of SDF-1 / CXCR4. It may be used in the treatment of vascular restenosis.