氧化型低密度脂蛋白及阿托伐他汀对培养的人脐静脉内皮细胞血管紧张素转化酶2表达的影响
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广东省自然科学基金(06301193)


Effects of Oxidized Low Density Lipoprotein and Atorvastatin on Angiotensin-Converting Enzyme-2 in Human Umbilical Vein Endothelial Cells
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    摘要:

    目的探讨氧化型低密度脂蛋白对人血管内皮细胞血管紧张素转化酶2表达的影响及阿托伐他汀的干预作用。方法用正常人新鲜血浆制备氧化型低密度脂蛋白,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。分为空白对照组、不同浓度氧化型低密度脂蛋白组(终浓度分别为20、40及80 mg/L)、不同浓度阿托伐他汀组(1、5和10μmol/L)、不同时间组(40 mg/L氧化型低密度脂蛋白和5μmol/L阿托伐他汀分别刺激6、12和24 h)及混合刺激组(5μmol/L阿托伐他汀+40 mg/L氧化型低密度脂蛋白刺激24 h)。提取细胞总RNA及蛋白,逆转录聚合酶链反应检测血管紧张素转化酶2 mRNA的表达,免疫印迹法检测血管紧张素转化酶2蛋白的表达。结果氧化型低密度脂蛋白呈剂量和时间依赖性下调血管紧张素转化酶2 mRNA及蛋白的表达。与空白对照组比较,不同浓度氧化型低密度脂蛋白组血管紧张素转化酶2 mRNA的表达分别降低为73%、51%和33%,蛋白的表达降低为81%、50%及32%,不同浓度组间比较差异亦有显著性(p<0.01);氧化型低密度脂蛋白培养6、12和24 h时间组血管紧张素转化酶2 mRNA与对照组比较分别减少为66%、55%和50%,蛋白的表达与对照组比较降低为63%、53%及49%(p<0.01)。不同浓度阿托伐他汀及其培养不同时间组血管紧张素转化酶2 mRNA和蛋白的表达与对照组比较差异无统计学意义。阿托伐他汀能抑制氧化型低密度脂蛋白对血管紧张素转化酶2 mRNA和蛋白表达的下调,mRNA表达增加为1.63倍,蛋白表达增加为1.60倍(p<0.01)。结论氧化型低密度脂蛋白呈剂量和时间依赖性下调血管紧张素转化酶2 mRNA和蛋白的表达,阿托伐他汀能抑制这种作用。

    Abstract:

    Aim To investigate the effects of oxidized low density lipoprotein(ox-LDL) and atorvastatin on gene and protein expression of angiotensin-converting enzyme-2(ACE-2) in human umbilical vein endothelial cells(HUVEC).Methods HUVEC cultured with equal serum-free medium were used as control.Different concentration groups of ox-LDL and atorvastatin: HUVEC were treated with ox-LDL(20,40 and 80 mg/L) and atorvastatin(1,5 and 10 μmol/L) for 24 h,respectively.In a timecontrol experiment,HUVEC were treated with ox-LDL at final concentration of 40 mg/L and atorvastatin of 5 μmol/L for 6,12 and 24 h,respectively.HUVEC were preincubated with atorvastatin(5 μmol/L) for 1 h and then cocultured with ox-LDL(40 mg/L) for 24 h.Changes in both gene and protein expression of ACE-2 were determined by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot analysis technique,respectively.Results Ox-LDL induced a concentration and time dependent decrease in gene and protein expression of ACE-2.When compared with that of control group,ACE-2 mRNA expression decreased to 73%,51%,33% and protein expression decreased to 81%,50% and 32%,respectively,after treated with different concentrations of ox-LDL(p< 0.01).When treated with ox-LDL for 6,12 and 24 h at concentration of 40 mg/L ox-LDL,ACE-2 mRNA expression in HUVEC decreased to 66%,55% and 50% when compared with that of control group( p<0.01),and protein expression decreased to 63%,53% and 49%(p< 0.01),respectively.Gene expression and protein expression of ACE-2 in groups treated with atorvastatin alone showed no significant difference when compared with that of control groups( p> 0.05).Pretreatment of HUVEC with simvastatin(5 μmol/L) for 1 h could obviously inhibited 40 mg/L ox

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周鹏,李志木梁,徐春生,严全能,陈文忠,范智文.氧化型低密度脂蛋白及阿托伐他汀对培养的人脐静脉内皮细胞血管紧张素转化酶2表达的影响[J].中国动脉硬化杂志,2007,15(12):889~892.

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  • 收稿日期:2007-08-27
  • 最后修改日期:2007-12-01
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