摘要
目的 评价缺氧预处理(HPC)对在缺氧状态下人脐静脉血管内皮细胞(HUVECs)细胞水平(细胞形态学变化、细胞活力、细胞凋亡率)、分子水平[细胞内血管内皮生长因子(VEGF)和血管生成素2(Ang-2)表达]及基因水平(相应mRNA的表达)的影响,探讨缺氧预处理对HUVECs缺氧损伤的保护作用机制.方法 常规进行人脐静脉血管内皮细胞株的培养,建立HUVECs缺氧模型,分为3组:正常对照(CON)组、缺氧(HYP)组及HPC组.在显微镜下观察各组细胞形态,分别采用CCK-8法、Hoechst 33258荧光染色和Western blotting法、RT-PCR法检测各组细胞活性、细胞凋亡率和细胞内VEGF、Ang-2及相应mRNA的表达水平.结果 与CON组相比,HYP组细胞明显受损、细胞活性明显减低,细胞凋亡率明显增加,细胞内VEGF、Ang-2及相应mRNA水平明显升高;而HPC组细胞形态接近正常,细胞活性及细胞凋亡率较CON组减低、但较HYP组高,细胞内VEGF、Ang-2及相应mRNA的表达水平较CON组及HYP组明显升高.结论 HPC通过上调细胞内VEGF、Ang-2及相应mRNA的表达对VEC缺氧损伤起到保护作用.
Abstract
Objective To investigate the effect of hypoxic preconditioning(HPC)on human umbilical vein endothelial cells(HUVECs)at the cellular level(cell morphology,viability,and apoptosis rate),the molecular level[the expression of vascular endothelial growth factor(VEGF)and angiogenin-2(Ang-2)],and the genetic level(the expression of corresponding mRNAs),and to discuss the protective mechanism of HPC against hypoxic injury of HUVECs.Methods HUVECs were routinely cultured to establish a hypoxia model of HUVECs,and then HUVECs were divided into normal control group(CON group),hypoxia group(HYP group),and HPC group.Cell morphology was observed for each group under a microscope,and CCK-8 assay,Hoechst33258 fluorescent staining,Western blotting,and RT-PCR were used to measure cell viability,cell apoptosis rate,and the expression levels of VEGF,Ang-2,and corresponding mRNAs in cells.Results Compared with the CON group,the HYP group had significant damage of cells,a significant reduction in cell viability,and significant increases in cell apoptosis rate and the levels of VEGF,Ang-2,and corresponding mRNAs in cells,while the HPC group had basically normal cell morphology,significantly lower cell viability and apoptosis rate than the CON group,and significantly higher cell viability and apoptosis rate than the HYP group,as well as significantly higher expression levels of VEGF,Ang-2,and corresponding mRNAs than the CON group and the HYP group.Conclusion HPC exerts a protective effect against hypoxic injury of HUVECs by upregulating the expression of VEGF,Ang-2,and corresponding mRNAs.
维普
万方数据