高血糖“代谢记忆”效应对血管内皮细胞损伤机制的研究-study on the mechanism of vascular endothelial cell injury caused by.docxVIP

华中科技大学硕士学位论文华中科技大学硕士学位论文33Study on the machanism of metabolic memory of hyperglycemia on injury of endothelial cellsABSTRACTObjective: to study the role and significance of transient High glucose on human vascular endothelial cells.Methods: Cultured primary human umbilical vein endothelial cells were used to study.HUVECs were divided into 4 group:1.Normal control group(NG,5.5mM D-glucose for 7 days),2.Persistent high glucose group(HG,30mM D-glucose for 7 days),3.Transient high glucose group(TG,30mM D-glucose for 1 day then 5.5mM D-glucose for 6 days),4.mannitol control group(MA,5.5mM D-glucose and 24.5mM D-mannitol for 7 days).We measured cell proliferation(MTT method),cell apoptosis(annexin V—FITC/PI double staining),ROS generation(DCFH-DA staining),SOD level(wst-1 method),MDAlevel(TBA method),NO level（Griess method）,eNOS mRNA(real-time PCR) and proteinlevel(western blot).Results: Cell proliferation is inhibited in HG group and TG group relative to NG group and also inhibited in MA group. High glucose can induce apoptosis of endothelial cells,incease ROS and MDA generation and down-regulate intracellular SOD level ,which can still maintain after re-establishing good blood glucose control. eNOS expression and NO generation of high glucose treated HUVEC persistent lower than NG group after a pe- riod of increasing when measured after the first 24h.Conclusion: Transient high glucose can inhibit endothelial cells proliferation and increase cell apoptosis,induce oxidative stress and reduce endothelium-dependent vasodilatation which can still persisitent even after re-establishment of good blood glucose control.Key words: Diabetes mellitus, diabetic complications, oxidative stress Nitric oxide, en- dothelial Nitrix oxide synthase11英文缩略词表英文缩写英文全称中文全称HUVEChuman umbilical vein endothelial cells人脐静脉内皮细胞ROSreactive oxygen species活性氧族MDAMalondialdehyde丙二醛SODsuperoxide dismutase超氧化物歧化酶NONitric oxide一氧化氮AGEsAdvanced glycation end products晚期高级糖基化终末产物FBSfetal bovine serum胎牛血清eNOSE