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目的研究腺病毒介导的线粒体融合素基因2(mitofusin2gene,Mfn2)对大鼠颈动脉球囊损伤后血管平滑肌细胞(vascularsmoothmusclecells,VSMCs)凋亡的影响。方法建立大鼠颈动脉球囊损伤模型,用携带大鼠Mfn2基因(rMfn2)的重组腺病毒Adv rMfn2GFP和含有绿色荧光蛋白基因的对照腺病毒Adv GFP分别感染球囊损伤动脉段,以磷酸缓冲液(PBS)作为未感染对照组,假手术组作为正常对照组,采用免疫组织化学方法检测外源基因表达水平;末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测凋亡细胞;Image Pro图像分析系统进行血管定量组织形态学分析。结果病毒感染后5d,免疫组织化学证实Adv rMfn2GFP组Mfn2蛋白表达水平明显;TUNEL染色显示,Adv rMfn2GFP组的VSMCs凋亡率明显高于PBS组和Adv GFP组,假手术组未见TUNEL阳性VSMCs(n=10,P<0.01);感染后21d,Adv rMfn2GFP组的血管内膜面积及内膜中膜面积比明显低于对照组(n=10,P<0.01)。结论高表达Mfn2基因可诱导大鼠颈动脉球囊损伤后血管平滑肌细胞凋亡。
Objective To investigate the effect of adenovirus-mediated mitofusin-2 gene (Mfn2) on the apoptosis of vascular smooth muscle cells (VSMCs) after carotid artery balloon injury in rats. Methods The model of carotid artery balloon injury in rats was established. The injured rabbit arteries were infected with recombinant adenovirus Adv rMfn2GFP carrying rat Mfn2 gene (rMfn2) and control adenovirus Adv GFP containing green fluorescent protein (EGFP) (PBS) as the uninfected control group and the sham operation group as the normal control group. The expression of exogenous gene was detected by immunohistochemical method; apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) Cells; Image Pro image analysis system for vascular quantitative histomorphological analysis. Results The expression of Mfn2 protein in Adv rMfn2GFP group was confirmed by immunohistochemistry 5 days after virus infection. The apoptosis rate of VSMCs in Adv rMfn2GFP group was significantly higher than that in PBS group and Adv GFP group by TUNEL staining, but not in TUNEL positive VSMCs group n = 10, P <0.01). At 21 days after infection, the intimal area and intima-media area ratio in Adv rMfn2GFP group were significantly lower than those in control group (n = 10, P <0.01). Conclusion High expression of Mfn2 gene can induce apoptosis of vascular smooth muscle cells after carotid artery balloon injury in rats.