缺氧条件下血管活性因子对血管内皮细胞中VEGF表达的影响

Effects of Vascular Activators on Expression of VEGF Gene in Human Umbilical Vein Endothelial Cells under Hypoxia

探讨在缺氧条件下人脐静脉血管内皮细胞对血管内皮生长因子 (vascular endothelialgrowth factor,VEGF)表达及缩血管活性物质内皮素 (ET)、舒血管活性物质一氧化氮 (NO)和 NO抑制剂 LNNA对 VEGF基因表达的影响 .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象分析等观察 VEGF m RNA和蛋白表达水平 .发现缺氧 6h内皮细胞可见 VEGF表达 .ET可促进 VEGF m RNA的表达 ,NO可明显抑制 VEGFm RNA的表达 ,NO抑制剂 LNNA也影响 VEGF m RNA的表达 .ELISA检测 VEGF蛋白水平分别为 6h组 8.2± 1 .1 ng/ L,ET+6h组 9.37± 1 .0 2 ng/ L,NO+6h组 2 .86± 0 .91 ng/ L,L - NNA+6h组 1 4.75± 1 .87ng/ L.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ET和 NO的调控 ,ET促进其表达 ,NO抑制其表达 .

To investigate the regulation and expression of VEGF gene in human umbilical vein endothelial cells under hypoxia and vascular activators,such as endothelin(ET),nitric oxide (NO) and NO synthesis inhibitor N\|nitro\| L \|arginine(L\|NNA),the human umbilical vein endothelial cells were cultured and then treated by hypoxia and vascular activators.The expression on VEGF mRNA level and protein level were assayed by Northern blots,ELISA and computer figure analysis system respectively.When human umbilical vein endothelial cells were subjected to hypoxia for 6 hours the expression of VEGF begans to increase.when vascular activators were added to the 6\|hour group,ET could increase inducement and NO inhibited it and L\|NNA stimulated the cells to express the VEGF.VEGF protein concentration in the cell culture media matched the mRNA result.They were 6 hours hypoxia 8.2±1.1 ng/L,ET+6 hours hypoxia 9.37±1.02 ng/L,NO+6 hours hypoxia 2.86±0.91 ng/L,L\|NNA+6 hours hypoxia 14.75±1,87 ng/L respectively.It was found that hypoxia could induce the expression of VEGF in human umbilical vein endothelial cells; ET could increase the inducible effect of hypoxia,NO could inhibit it and LNNA stimulated the expression of VEGF in human umbilical vein endothelial cells.