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目的研究血管内皮生长因子C(VEGF-C)、基质细胞衍生因子-1(SDF-1)、单核细胞趋化蛋白-1(MCP-1)和粒细胞集落刺激因子(G-CSF)对CD34+/CD133+/VEGFR-3+淋巴管内皮祖细胞(LEPCs)的趋化和动员作用。方法用Percoll非连续密度梯度离心法分离狗外周血单个核细胞,再用流式细胞术分选VEGFR-3+LEPCs,激光扫描共焦显微镜下观察细胞特征性标志物CD34和CD133的表达。通过跨膜迁移实验观察VEGF-C、SDF-1、MCP-1和G-CSF对LEPCs的趋化作用,并用扫描电镜观察迁移细胞的形态特征。在大鼠皮下分别注射VEGF-C、SDF-1、MCP-1和G-CSF,用流式细胞术检测外周血单个核细胞中LEPCs数目,同时用全自动血样分析仪计数白细胞。结果与对照组相比,VEGF-C、SDF-1、MCP-1和G-CSF组跨膜迁移细胞的百分率增大,用VEGFR-3和CXCR-4阻断抗体处理后VEGF-C和SDF-1的趋化迁移作用明显降低。注射VEGF-C、SDF-1、MCP-1和G-CSF后,大鼠外周血中LEPCs的数量明显增多。未见白细胞显著增多。结论VEGF-C、SDF-1、MCP-1和G-CSF对LEPCs有趋化迁移和动员作用,VEGF-C/VEGFR-3和SDF-1/CXCR-4信号途径在LEPCs趋化迁移方面起着重要调控作用。
Objective To investigate the effects of vascular endothelial growth factor C (VEGF-C), stromal cell-derived factor-1 (SDF-1), monocyte chemoattractant protein- 1 (MCP-1) and granulocyte colony stimulating factor Chemotaxis and mobilization of CD34 + / CD133 + / VEGFR-3 + lymphatic endothelial progenitor cells (LEPCs). Methods Peripheral blood mononuclear cells were isolated by Percoll non-continuous density gradient centrifugation. VEGFR-3 + LEPCs were separated by flow cytometry. The expression of CD34 and CD133 were observed under laser scanning confocal microscope. The chemotactic effects of VEGF-C, SDF-1, MCP-1 and G-CSF on LEPCs were observed by transmembrane migration assay. Morphological characteristics of migrating cells were observed by scanning electron microscopy. The rats were subcutaneously injected with VEGF-C, SDF-1, MCP-1 and G-CSF respectively. The number of LEPCs in peripheral blood mononuclear cells was detected by flow cytometry, and the white blood cells were counted by automatic blood sample analyzer. Results Compared with the control group, the percentage of transmembrane migration in VEGF-C, SDF-1, MCP-1 and G-CSF groups increased. VEGF-C and SDF were blocked by VEGFR-3 and CXCR- -1 chemoattractive effect was significantly reduced. After injection of VEGF-C, SDF-1, MCP-1 and G-CSF, the number of LEPCs in peripheral blood of rats increased significantly. No significant increase in white blood cells. Conclusions VEGF-C, SDF-1, MCP-1 and G-CSF have chemotactic and mobilization effects on LEPCs. VEGF-C / VEGFR-3 and SDF-1 / CXCR-4 signaling pathways play an important role in chemotactic migration of LEPCs An important role in regulation and control.