The role of eNOS in the migration and proliferation of bone-marrow derived endothelial progenitor cells and in vitro angiogenesis
The role of eNOS in the migration and proliferation of bone-marrow derived endothelial progenitor cells and in vitro angiogenesis
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eNOS在骨髓源性内皮祖细胞迁移和增殖及体外血管生成中的作用
DOI：
10.1002/cbin.10405
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发表时间：
2015-04-01
影响因子：
3.9
通讯作者：
Qian, Liling
中科院分区：
文献类型：
--
作者：
Lu, Aizhen;Wang, Libo;Qian, Liling
研究了内皮型一氧化氮合酶(ENOS)在体外内皮祖细胞(EPC)迁移、增殖和管状形成中的作用。大鼠骨髓单个核细胞在EGM-2 MV中培养7-10天获得内皮祖细胞，并通过荧光显微镜观察其结合FITC-UEA-1的能力和摄取乙酰化低密度脂蛋白(Dil-Ac-LDL)的能力进行鉴定。在eNOS抑制剂N-硝基-L-精氨酸甲酯(L-NAME)存在或不存在的情况下，检测细胞的迁移、增殖和管形成活性。用实时荧光定量聚合酶链式反应和免疫印迹法检测细胞中CXCR4、CXCR7、VEGFR2和eNOSmRNA和蛋白的表达。用硝酸还原酶检测L存在或不存在时一氧化氮的产生。第7~10天出现典型的梭形细胞，融合达80%左右。FITC-UEA-1和Dil-Ac-LDL双染细胞的比例约为85%。ENOS被抑制后，细胞的迁移、增殖和管状形成明显减弱(P
The role of endothelial nitric oxide synthase (eNOS) in the activities of endothelial progenitor cells (EPCs) including migration, proliferation, and tube formation in vitro was investigated. EPCs were obtained from rat bone mononuclear cells by culturing for 7-10 days in EGM-2MV and identified by their capacity for FITC-UEA-1 binding and acetylated low-density lipoprotein (Dil-ac-LDL) intake using fluorescence microscopy. Migration, proliferation and tube formation activities were assessed in the presence or absence of N-nitro-L-argininemethylester (L-NAME), an eNOS inhibitor. mRNA and protein expression of CXCR4, CXCR7, VEGFR2, and eNOS were detected by real-time PCR and western blotting in the presence or absence of L-NAME. Nitric oxide production was detected by nitrate reductase in the presence or absence of L-NAME. Typical spindle-shaped cells appeared on the 7(th)-10(th) day and confluence reached about 80%. The percentage of FITC-UEA-1 and Dil-ac-LDL double-stained cells was about 85%. Cell migration, proliferation, and tube formation were significantly weakened after eNOS was inhibited (P