腺病毒介导的eNOS基因转移对血管平滑肌细胞增殖的抑制作用

目的 :探讨腺病毒载体介导的内皮型一氧化氮合酶 (eNOS)基因转染血管平滑肌细胞 (VSMC)的效率及其对冠状动脉旁路移植术后再狭窄基因治疗的可行性。方法 :构建带有eNOS基因的复制缺陷型重组腺病毒转染体外培养的VSMC ,应用聚合酶链 (PCR)、逆转录PCR(RT PCR)技术和免疫组化法检测外源基因转染及表达效率 ;应用MTT法和流式细胞仪检测eNOS基因对VSMC的抑制作用。结果 :外源性eNOS基因成功导入了VSMC ;VSMC中有eNOS基因mRNA的表达 ;外源性eNOS蛋白呈高效表达。eNOS基因重组腺病毒载体转染细胞后 ,可显著抑制VSMC的生长 ,DNA合成减少。结论 :腺病毒介导的eNOS基因可高效地转染VSMC ,并可有效抑制细胞生长。

Adenovirus-mediated endothelial nitric oxide synthase gene transfer inhibits vascular smooth muscle cells proliferation

Objective:To study the transfection efficiency of adenovirus mediated endothelial nitric oxide synthase (eNOS) gene and the possibility of eNOS gene in prevention of restenosis after coronary artery bypass graft (CABG).Methods:A replication defective adenovirus encoding eNOS (AdCMVeNOS) gene was correctly constructed and transfected into vascular smooth muscle cells (VSMC) in vitro. The efficiency of transfection and expression was detected by polymerase chain reaction (PCR) techniques and RT PCR and immunohistochemistry. The inhibition of AdCMVeNOS on the proliferation of VSMCs was detected by MTT and flow cytometric tests.Results:The eNOS cDNA ,mRNA and protein could be detected within the transfected cells. The efficiency of transfection and expression was high. The proliferation and DNA synthesis of VSMCs could be inhibited by the transferred eNOS gene.Conclusion:eNOS gene could be quickly and effectively transferred into VSMCs by adenovirus vector. AdCMVeNOS might play a role in prevention of restenosis.