ACE2基因表达增加对人脐静脉内皮细胞增殖、凋亡的影响

The effect of ACE2 gene overexpression on proliferation and apoptosis of human umbilical vein endothelial cells induced by Angiotensin lI

目的探索在AngII干预下ACE2基因表达增加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。用构建、包装好的慢病毒重组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（HUVEC）。感染72h后，以AnglI（终浓度为10。mol／L）干预细胞，倒置相差显微镜观察各组HUVEC的形态学变化，AlamarBlue试剂检测各组HUVEC增殖功能，TUNEL细胞凋亡检测试剂盒检测各组HUVEC的凋亡。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；而正常细胞对照组与AngII＋Lentiviral—ACE2组细胞生长状态良好，圆形、卵圆形，饱满，形态正常呈“铺路石”样生长，紧密贴壁，悬浮细胞少。AngII组较正常细胞对照组、Ang1I＋Lentiviral—ACE2组AlamarBlue被还原率明显降低（P〈O．05）。Ang11组的凋亡指数为（0．1165±0．0181），与正常细胞对照组凋亡指数（0．0373±0．0113）和AngⅡ＋Lentiviral—ACE2组凋亡指数（0．0540±0．0061）相比差异有统计学意义（P〈0．05）。AngII组与AngⅡ＋Lentiviral—GFP相比、正常细胞对照组与AngII＋Lentiviral—ACE2组相比，AlamarBlue被还原率和凋亡指数差异无统计学意义。结论AngⅡ具有促进人脐静脉内皮细胞增殖能力下降和凋亡增加的作用。ACE2表达增~IIII抑制AngⅡ诱导的人脐静脉内皮细朐增殖活件降低和凋亡增加．对人脐静脉内皮细胞具有保护作用。

Objective To study the effect of ACE2 gene overexpression on proliferation and apoptosis in- duced by angiotensin H in human umbilical vein endothelial cells. Methods Human umbilical endothelial cells were cultured in vitro. The human umbilical vein endothelial cells were infected by recombinant ACE2 gene lentiviral vector （Lentiviral-ACE2） with multiplicity of infection of 10 （MOI=IO） which had been constructed, packaged. After infected by Lentiviral-ACE2 72 hours. The human umbilical vein endothelial cells were treated with Ang II （at final concentration 10-7 mol/L）. The morphological changes of human umbilical vein endothelial cells in each group was observed by inverted phase contrast microscope. The proliferation function of each group human umbilical vein endothelial cells was detected by AlamarBlue reagents. The apoptosis in each group human umbilical vein endothelial cells was researched by TUNEL Apoptosis Detection Kit. Results The cell growth state in Ang !1 group and Ang !1 ＋ Lentiviral-GFP group were poor, slow growth, irregular shape and there were some degrees of cell suspension off wall, while the normal cell control group and group of Ang I1 ＋Lentiviral-ACE2 cell growth were in good condition, round, oval, full, and they were ＂paving stone＂-like normal morphology growth and closely adherent, having little suspension cells. Ang I1 group＇ AlamarBlue reduction rate was significantly lower （P〈0.05） than those of normal cell control group and Ang Ill ＋Lentiviral-ACE2 group. The apoptosis indexwas also a statistical significance （P〈0.05） in Ang ]I group （0.1165±0.0181） compared to normal cell control group（0.0373±0.0113） and Ang l] ＋Lentiviral-ACE2 group （0.0540±0.0061）. The AlamarBlue reduction rate and apoptosis index between Ang II group and Ang II ＋Lentiviral-GFP was not statistically significant, nor between normal cells control group and Ang lI ＋Lentiviral-ACE2. Conclusion Ang 11 has a function that it can reduce the proliferation and increase the apoptosis of human umbilical vein endothelial cells,while the overexpression of ACE2 gene can restrain those actions.