eNOS对大鼠颈动脉球囊损伤后平滑肌细胞增殖的影响

目的研究内皮型一氧化氮合酶(eNOS)对大鼠球囊损伤后平滑肌细胞增殖的影响。方法构建腺病毒-eNOS(AdCMV-eNOS)质粒,以脂质体介导转染293细胞,扩增后转染SD大鼠球囊损伤后平滑肌细胞(实验组),以转染Ad-LacZ重组质粒的平滑肌细胞作为对照组。RT-PCR技术观察转染后平滑肌细胞eNOS mRNA的表达;细胞计数法分别检测实验组、对照组、正常组和单纯球囊损伤组平滑肌细胞的增殖能力。。结果RT-PCR检测发现实验组平滑肌细胞表达eNOS mRNA,而对照组无表达。各时间点细胞计数结果显示,正常组均明显少于单纯球囊损伤组(P<0.05),而实验组均明显少于单纯球囊损伤组和对照组(P<0.05)。结论重组腺病毒表达载体构建正确;eNOS可以抑制平滑肌细胞增殖。为eNOS预防经皮腔内冠状动脉成形术术后再狭窄基因转染奠定了基础。

Effect of endothelial nitric oxide synthase on proliferation of rat vascular smooth muscle cells from ballon-injured carotid artery

Objective To study the effect of endothelial nitric oxide synthase(eNOS) on proliferation of rat vascular smooth muscle cells from ballon-injured carotid artery. Methods An adenovirus expression vector for eNOS(AdCMV-eNOS) was constructed and propagated in HEK293 cells by Fugene 6.AdCMV-eNOS was transfected into SD-rat vascular smooth muscle cells from ballon-injured carotid artery(experiment group).Smoth muscle cells transfected with Ad-LacZ recombinant plasmid were served as control group.Cell counting was performed to observe the effect on cell proliferation in experiment group,control group,normal group and simple ball on injury group.While RT-PCR was employed to examine the eNOS mRNA expression. Results It was revealed by RT-PCR that the smooth muscle cells in the experiment group expressed eNOS mRNA,while those in the control group did not.Cell counting indicated that the normal group was significantly less than the simple ballon injury group at each time points(P<0.05),while the experiment group was significantly less than the simple ballon injury group and control group(P<0.05). Conclusion The recombinant adenovirus expression vector can be correctly constructed,and eNOS can inhibite the proliferation of vascular smooth muscle cells,laying a foundation for eNOS transfection in the prevention of postangioplasty restenosis.