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ACE2对脂多糖损伤人脐静脉内皮细胞的抗黏附作用

, PP. 1302-1307

Keywords: 脂多糖,ACE,脐静脉内皮细胞,单核细胞

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Abstract:

目的观察脂多糖(lipopolysaccharide,LPS)诱导血管内皮细胞损伤后细胞黏附性改变与血管紧张素转化酶2(angiotensin-convertingenzyme2,ACE2)的表达变化的关系,探讨ACE2对血管内皮细胞损伤所致黏附性增加的对抗作用。方法分别用不同浓度(0、10、100、1000ng/mL)LPS诱导人脐静脉内皮细胞(umbilicalveinendothelialcell,HUVEC)致其损伤,人单核细胞白血病细胞(humanacutemonocyticleukemiacellline,THP-1)黏附实验观察HUVEC的黏附性改变。本实验设置4个实验组,分别为未经任何处理的空白对照组、1000ng/mLLPS处理6h的模型组、以10μmol/L氯沙坦钾预处理模型组的氯沙坦钾组及以10μmol/LA779预处理空白对照组的A779组(n=3)。Westernblot及RT-PCR法分别检测ACE2的蛋白和基因表达,ELISA法检测细胞上清中血管紧张素Ⅱ(AngⅡ)、血管紧张素1-7(Ang1-7)及细胞间黏附分子1(ICAM-1)的含量。结果单核细胞黏附实验可见黏附在HUVEC上的THP-1呈绿色荧光,黏附细胞数呈LPS诱导浓度及时间依赖性显著增加(P<0.01);Westernblot结果显示,LPS诱导HUVEC损伤模型组中ACE2蛋白表达水平较空白对照组显著减少(37.33±3.50)%(P<0.01);RT-PCR结果显示,模型组中ACE2mRNA表达水平较空白对照组显著减少(26.31±2.19)%(P<0.01);ELISA结果显示,模型组较空白对照组细胞上清Ang1-7显著降低(P<0.05),AngⅡ及ICAM-1均显著升高(P<0.01),氯沙坦钾处理组较模型组ICAM-1显著降低(P<0.01),A779处理组较空白对照组ICAM-1显著升高(P<0.05)。结论LPS诱导损伤HUVEC细胞后其黏附性增加与ACE2表达下调密切相关,提示ACE2具有一定抗黏附作用。

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