摘要
目的:观察腺病毒介导的反义AT1基因转染对培养的大鼠动脉平滑肌细胞(VSMCs)迁移的影响。方法:用定向克隆和同源重组方法构建携带人反义AT1基因的复制缺陷型重组腺病毒(AdCMV/ahAT1),转染体外培养的VSMCs,用RT-PCR半定量法和免疫组化法检测AT1R的表达,用改良趋化小室法检测细胞迁移。结果:与对照组相比转染AdCMV/ahAT1后48h的VSMCs,AT1RmRNA表达少50％,AT1R的蛋白表达也显著低下(P<0.01);迁移距离为(41.7±9.7)μm,显著短于空白对照组(77.5±12.7)μm和Ad/CMV.LacZ组(77.2±10.6)μm,均为P<0.01。结论:腺病毒介导的反义hAT1R转染,通过抑制AT1R的表达,明显抑制了大鼠VSMCs的迁移。
AIM: To evaluate the role of human angiotensin Ⅱ(AngⅡ) type Ⅰ receptor (AT1R) antisense cDNA (ahAT1) on migration of cultured artery smooth muscle cells (VSMCs). METHODS: Two recombinant adenoviral vectors, Ad/CMV. ahAT1 containing full length antisense cDNA targeting to human AT1R mRNA, and Ad/CMV.LacZ containing LacZ called report gene, were constructed by orientation clone technology and homologous recombination, and then were used to transfect VSMCs in vitro. AT1R expression detected by RT-PCR and immunohistochemistry, and migration of VSMCs measured by Boyden's Chamer methods, were compared between transfected and non-
transfected VSMCs. RESULTS: Forty-eight hours after Ad/CMV. ahAT1 transfection, the level of AT1R mRNA decreased markedly (50% of control group), and AT1R protein expression was significantly less (P<0.01 vs control group and Ad/CMV. LacZ-group, respectively) in VSMCs. So it was migration distance of VSMCs among the three groups. CONCLUSION: These results indicate that antisense cDNA targeting to human AT1R transfer in vitro mediated by adenoviral vector has a powerful inhibitory effect on migration of VSMCs by attenuating AT1R expression.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第3期429-432,共4页
Chinese Journal of Pathophysiology
基金
湖北省自然科学基金(No.2001ABB167)
卫生厅项目(No.WJ01580)
关键词
血管紧张素Ⅱ
腺病毒
人
肌
平滑
血管
细胞运动
Angiotensin Ⅱ
Adenoviruses
human
Muscle
smooth
vascular
Cell movement
