Title: Effects of adenoviral vector containing human AT1 receptor antisense cDNA on the proliferation and apoptosis of human vascular smooth muscle cells
Abstract: To identify the role of human angiotensin Ⅱ (AngⅡ) type 1 receptor (AT1R) antisense cDNA (ahAT1) on the proliferation and apoptosis of cultured human artery tissue smooth muscle cells (VSMCs) induced by AngⅡ, Two recombinant adenoviral vectors, Ad/CMV.ahAT1 containing full length antisense cDNA targeting to human AT1R mRNA, and Ad/CMV.LacZ containing LacZ (report gene) were constructed by orientation clone technique and homologous recombination, and then transfected into VSMCs from human umbilical artery. After stimulation for 24h by AngⅡ 10 -7 mol/L, AT1R expression was detected by RT-PCR and immunohistochemical method, Proliferation index (PI) and apoptosis of VSMCs were determined by flow cytometry. Ad/CMV.LacZ transfected and nontransfected VSMCs served as control. Forty eight hours after being transfected Ad/CMV.ahAT1 into VSMCs, the level of AT1R mRNA decreased markedly (50% of control-group) and AT1R protein expression was significantly less than that in other groups (P0.01 compare with control-group and Ad/CMV.LacZ-group respectively). AngⅡ markedly increased PI of VSMCs in a time-dependent manner (P0.05 or P0.01 compare with control). The increased PI of VSMCs was blocked to control level by transfecting Ad/CMV.ahAT1 into VSMCs named as Group AngⅡ-Ad/CMV.ahAT1 (P0.05 compare with control, 0.01 compare with AngⅡ stimulated only). But there was no statistic difference in these parameters between Group AngⅡ-Ad/CMV.LacZ and Group AngⅡ. Apoptosis peak emerged only in Group AngⅡ-Ad/CMV.ahAT1. The rate of apoptosis in those VSMCs used Ad/CMV.ahAT1 and AngⅡ was 1% to 28%. These results indicated that antisense cDNA targeted to human AT1R transfer in vitro mediated by adenoviral vector had a powerful inhibitory effect on AngⅡ-induced proliferation of VSMCs by attenuating AT1R expression.
Publication Year: 2004
Publication Date: 2004-01-01
Language: en
Type: article
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