氧化高密度脂蛋白对培养的人脐静脉内皮细胞血管紧张素转化酶表达的影响  

Effect of oxidized High density lipoprotein on angiotensinConverting Enzyme of human vascular endothelial cell.

作　　者：李晓庆[1] 王秋林[1] 张磊[1] 姜轶[1] 

机构地区：[1]成都医学院第一附属医院,四川成都610500

出　　处：《四川医学》2008年第7期805-808,共4页Sichuan Medical Journal

基　　金：四川省教育厅自然科学科研项目(No:2006B109)

摘　　要：目的探讨氧化高密度脂蛋白（ox-HDL）对人血管内皮细胞血管紧张素转化酶（ACE）表达的影响。方法人脐静脉内皮细胞株复苏、传代后取生长良好3～6代用于实验。分为①空白对照组：加无血清培养基;②不同浓度组：ox-HDL（终浓度分别为20、40、80mg/L）与人脐静脉内皮细胞孵育24h;③不同时间组：以终浓度为40mg/L氧化型高密度脂蛋白分别刺激6、12和24h。提取细胞总RNA及蛋白,逆转录聚合酶链反应（RT-PCR）检测血管紧张素转化酶mRNA表达,以蛋白质免疫印迹法（Western blot）检测ACE蛋白表达量。结果ox-HDL呈剂量（浓度）和时间依赖性使ACE基因及蛋白表达量增加,与对照组比较,ox-HDL各浓度刺激组ACE mRNA表达增加1.43、2.41、2.98倍,ACE蛋白表达增加为1.32、2.0、2.54倍（P均〈0.01）,不同浓度组间比较差异亦有统计学意义（P〈0.01）。于6、12和24h,ACE mRNA表达增加1.64、2.08、2.43倍,ACE蛋白表达增加1.52、1.78、1.99倍（P均〈0.01）,不同时间组间比较差异有统计学意义（P〈0.05）。结论ox-HDL呈剂量（浓度）和时间依赖性显著上调ACE基因及蛋白表达,并通过这一机制参与动脉粥样硬化形成。Objective To investigate the effect of oxidized high density lipoprotein（ox-HDL） on the gene and protein expression of angiotensin-converting enzyme（ACE） in human umbilical vein endothelial cells（HUVECs）.Methods Pproducing ox-HDL from human fresh plasma,HUVECs of 3～6th passage were rolled into the experiment and divided into three groups：（1）cells cultured with equivalence Serum-free medium as control;（2）Various concentrations groups of ox-HDL：HUVECs were treated with ox-HDL（20mg/L、40mg/L、80mg/L） for 24 hours;（3） In a time-control experiment,HUVECs were treated with ox-HDL at the final concentration of 40mg/L for 6,12 and 24 hours,respectively.Changes in both gene and protein expression of ACE were determined with reverse transcription polymerase chain reaction RT-PCR） and Western blot analysis technique,respectively.Results ox-HDL induced a concentration and time dependent increase in both gene and protein expression of ACE.Compared with control group,treatment of HUVECs with ox-HDL at the concentration of 20、40、80mg/L stimulated 1.43、2.41 and 2.98 fold increase in the ACE mRNA expression,As well As 1.32、2.0 and 2.54 fold increase in their protein expression,respectively.After treatment of HUVECs with ox-HDL for 6,12 and 24 hours at the concentration of 40mg/L,the ACE gene expression increased 1.64、2.08 and 2.43 fold,and its protein expression increased 1.52、1.78 and 1.99 fold;respectively.Conclusion ox-HDL up-regulates the protein and gene expression of ACE in HUVECs in a concentration and time dependent manner.

关 键 词：氧化高密度脂蛋白 血管紧张素转化酶 人脐静脉内皮细胞 

分 类 号：R363[医药卫生—病理学]