血管紧张素转化酶2基因过表达改善人脐静脉内皮细胞生长

摘要

Objective To explore the effects of overexpression of angiotensin-convrting enzyme 2 (ACE2) on the proliferation and apoptosis induced by angiotensin Ⅱ in human umbilical vein endothelial cells (HUVECs). Methods In vitro cultured HUVECs were infected with recombinant lentiviruses expressing ACE2 (Lentiviral-ACE2) for 72 hours with an multiplicity of infection of 10 (MOI=10); HUVECs were then treated with Ang Ⅱ at a final concentration of 10-7 mol/L . The morphological changes of HUVECs in each group were observed by inverted phase contrast microscope ; the proliferation of HUVEC in each group was detected by Alamar Blue reagents ; and the apoptosis in each group was assayed by TUNEL Apoptosis Detection Kit . Results Compared to untreated normal control cells and Ang II + Lentiviral-ACE2 cells , cells in Ang Ⅱ group and Ang Ⅱ + Lentiviral-GFP group were in poor growing conditions with slow doubling time , irregular cell morphologies ; lots cells were floating in the culture medium ; Alamar Blue reduction rate in Ang II group was significantly lower (P<0 .05) than those of normal cell control group and Ang II +Lentiviral-ACE2 group . Apoptosis index (0 .12 ± 0 .02) in Ang Ⅱ group was also statistical significant higher (P<0 .05 ) compared to those in normal cell control group (0 .04 +0 .01) and AngⅡ + Lentiviral-ACE2 group(0 .05 + 0 .01) . However , the Alamar Blue reduction rate and apoptosis index between Ang Ⅱ group and Ang Ⅱ + Lentiviral-GFP and between normal cells control group and Ang Ⅱ +Lentiviral-ACE2 were not statistically significant different . Conclusion Ang Ⅱ reduces proliferation and increases apoptosis of human umbilical vein endothelial cells , while the overexpression of ACE2 protects Ang Ⅱ-treated HUVEC by increasing proliferation and decreasing apoptosis .%目的 探索在血管紧张素转化酶2(ACE2)基因过表达干预下血管紧张素(AngⅡ)对人血管内皮细胞增殖、凋亡的影响.方法 原代培养人脐静脉血管内皮细胞.慢病毒重组ACE2基因表达载体(Lentiviral-ACE2)以感染复数为10(MOI=10)感染人脐静脉内皮细胞(HUVECs).感染72 h后,以AngⅡ(终浓度为10-7mol/L)干预细胞,倒置相差显微镜观察HUVECs的形态学变化,AlamarBlue试剂检测HUVECs增殖功能,TUNEL法检测HUVECs凋亡.结果 AngⅡ组与AngⅡ+Lentiviral-GFP组细胞生长状态差,生长缓慢,形态不规则并且有不同程度脱壁悬浮的细胞;而正常细胞组与AngⅡ+Lentiviral-ACE2组细胞生长状态良好.AngⅡ组较正常细胞组、AngⅡ+Lentiviral-ACE2组AlamarBlue被还原率明显降低.AngⅡ组的凋亡指数(0.11±0.02)与正常细胞组(0.04±0.01)和AngⅡ+Lentiviral-ACE2(0.05±0.01)比较,具有统计学意义(P<0.05).AngⅡ组与AngⅡ+Lentiviral-GFP比较、正常细胞组与AngⅡ+Lentiviral-ACE2组比较,AlamarBlue被还原率和凋亡指数差别无统计学意义(P>0.05).结论 AngⅡ具有降低人脐静脉内皮细胞增殖能力和增加细胞凋亡的作用,ACE2过表达改善AngⅡ诱导的人脐静脉内皮细胞增殖和减少细胞凋亡,对人脐静脉内皮细胞具有保护作用.