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目的 探讨在缺氧条件下人脐静脉血管内皮细胞血管内皮生长因子 (VEGF)表达及缩血管活性物质内皮素 ,舒血管活性物质NO和NO抑制剂LNNA对VEGF基因表达的影响。方法 体外培养人脐静脉血管内皮细胞 ,经缺氧及血管活性物质处理 ,Northern杂交、酶联免疫检测和计算机图像分析等观察VEGFmRNA和蛋白表达水平。结果 缺氧 6h内皮细胞可见VEGF表达。ET可促进VEGFmRNA的表达 ,NO可明显抑制VEGFmRNA的表达 ,NO抑制剂LNNA也影响VEGFmRNA的表达。ELISA检测VEGF蛋白水平分别为 6h组 (8 2± 1 1) μg/L ,ET +6h组 (9 37± 1 0 2 ) μg/L ,NO +6h组 (2 86± 0 91) μg/L ,LNNA +6h组 (14 75± 1 87)μg/L。 结论 缺氧可诱导人脐静脉血管内皮细胞分泌VEGF并受血管活性物质的调控 ,ET促进其表达 ,NO抑制其表达。
Objective To investigate the effect of hypoxia on the expression of vascular endothelial growth factor (VEGF) and the vasoconstrictural endothelin (vasoactive endothelin), vasodilator NO (NO) and nitric oxide synthase (LNNA) in human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells were cultured in vitro. The expression of VEGF mRNA and protein was observed by hypoxia and vasoactive substances treatment, Northern blotting, enzyme-linked immunosorbent assay and computer image analysis. Results The expression of VEGF was observed in endothelial cells 6 h after hypoxia. ET can promote the expression of VEGFmRNA, NO can significantly inhibit the expression of VEGFmRNA, NO inhibitor LNNA also affect the expression of VEGFmRNA. The protein expression of VEGF in the group of 6 h (8 2 ± 1 1) μg / L, ET + 6h (9 37 ± 1 0 2) μg / L, and NO + 6h , LNNA + 6h group (14 75 ± 1 87) μg / L. Conclusions Hypoxia can induce the secretion of VEGF in human umbilical vein endothelial cells, which is regulated by vasoactive substances. ET can promote the expression of VEGF and NO can inhibit the expression of VEGF.