血管紧张素Ⅱ对外周血早期内皮祖细胞血管内皮生长因子表达的影响
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江苏大学临床医学科技发展基金资助项目(JLY20080035)


Effects of Angiotensin Ⅱ on Vascular Endothelial Growth Factor Expression in Early Endothelial Progenitor Cells from Human Peripheral Blood
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    摘要:

    目的观察血管紧张素Ⅱ对外周血早期内皮祖细胞血管内皮生长因子表达的影响。方法密度梯度离心法获取外周血单个核细胞,培养7天,收集贴壁细胞,随机分对照组、血管紧张素Ⅱ各浓度(10-3mol/L、10-5mol/L、10-7mol/L)组、血管紧张素Ⅱ+缬沙坦组、血管紧张素Ⅱ+PD123319组。多波长激光共聚焦显微镜鉴定FITC标记荆豆凝集素Ⅰ和D iI标记的乙酰化低密度脂蛋白双染色阳性为早期内皮祖细胞,流式细胞仪检测其表面标志进一步鉴定。酶联免疫吸附测定法对早期内皮祖细胞血管内皮生长因子的表达进行测定。结果血管紧张素Ⅱ呈浓度依赖方式上调早期内皮祖细胞血管内皮生长因子的表达,此作用可被缬沙坦显著抑制,使之接近正常水平,PD123319对此无明显作用。结论血管紧张素Ⅱ通过血管紧张素Ⅱ受体1介导上调早期内皮祖细胞的血管内皮生长因子的表达,并呈浓度依赖性,血管紧张素Ⅱ受体2不参与血管紧张素介导的血管内皮生长因子的分泌。

    Abstract:

    Aim To investigate the effect of angiotensin Ⅱ on vascular endothelial growth factor expression of early endothelial progenitor cells. Methods Total mononuclear cells(MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation,and then the cells were plated on fibronectin-coated culture dishes.After 7 days of culture,several groups of attached cells were incubated with angiotensin Ⅱ(to make a series of concentrations: 10-3 mol/L,10-5 mol/L,10-7 mol/L vehicle control for 24 h),angiotensin Ⅱ+valsartan,angiotensinⅡ+PD123319.The cells were observed under inverted microscope,and characterized as adherent cells double positive for DiL DL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope.The early endothelial progenitor cells were further documented by demonstrating the expression of cell markers with flow cytometry.Enzyme-linked immunospecific assay(ELISA) were used to assess vascular endothelial growth factor expression. Results Our data indicated that angiotensin Ⅱ can significantly increase the vascular endothelial growth factor expression,with a maximum at 10-3 mol/L after 24 hours(P<0.05);These effects can be attenuated by pre-treatment with valsartan but not PD123319. Conclusion It is suggested that angiotensin Ⅱ induces vascular endothelial growth factor protein secretion via the angiotensin Ⅱ receptor-1 but not angiotensin Ⅱ receptor-2.

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孙文文,任国庆,汪奕斌,张浩.血管紧张素Ⅱ对外周血早期内皮祖细胞血管内皮生长因子表达的影响[J].中国动脉硬化杂志,2011,19(8):651~654.

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  • 收稿日期:2010-12-23
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