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.
出处
《四川医学》
CAS
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)
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
oxidized high density lipoprotein
angiotensin-converting enzyme
human umbilical vein endothelial cells
