转染ANT1基因诱导颈动脉球囊损伤大鼠血管平滑肌细胞凋亡  

Adenine translocase 1 gene transfection induces apoptosis of vascular smooth muscle cells in rats with carotid balloon injury

作　　者：杨易[1] 李燕[1] 王霄[1] 刘洁[1] 卢巍[1] 宋耀明[1] 

机构地区：[1]第三军医大学新桥医院心血管内科,全军心血管病研究所,重庆400037

出　　处：《第三军医大学学报》2012年第8期738-743,共6页Journal of Third Military Medical University

摘　　要：目的通过腺病毒载体在体转染大鼠颈总动脉球囊损伤模型,研究过表达腺嘌呤核苷酸转位酶-1(adeninenucleotide translocase 1,ANT1)对血管平滑肌细胞凋亡的影响。方法将72只雄性SD大鼠随机分为正常组、单纯损伤组、Ad-GFP转染组、Ad-ANT1转染组,每组18只。用携带ANT1的腺病毒载体(Ad-ANT1)或空载体腺病毒载体(Ad-GFP)局部感染大鼠颈总动脉球囊损伤动物模型,在损伤后7、14、28 d,取大鼠损伤侧颈动脉RT-PCR检测ANT1 mRNA表达,West-ern blot以及免疫组化检测BAX和Bcl-2表达,TUNEL法原位检测新生内膜及中膜VSMCs凋亡率。结果 Ad-ANT1腺病毒转染后大鼠颈动脉ANT1大量表达,在14 d达到高峰,显著高于Ad-GFP转染组及单纯球囊损伤组(P<0.01),至28 d时仍有表达。在7、14 d时,Ad-ANT1转染组BAX阳性表达率显著高于Ad-GFP转染组及单纯损伤组(P<0.05);Bcl-2阳性表达率高于正常血管,但与Ad-GFP转染组和单纯损伤组无显著差异(P>0.05);Ad-ANT1转染组新生内膜及中膜平滑肌细胞凋亡率显著高于其他各组(P<0.05);在14、28 d时,Ad-ANT1转染组新生内膜及中膜面积比(IA/MA)小于Ad-GFP转染组及单纯损伤组(P<0.05)。结论腺病毒载体介导过表达ANT1可诱导大鼠颈总动脉球囊损伤动物模型新生内膜及中膜VSMC的凋亡,其作用可能通过上调促凋亡蛋白BAX的表达实现。Objective To study the effect of adenine nucleotide translocase 1 pression on apoptosis of vascular smooth muscle cells （VSMC） in a rat carotid balloon （ANT1） gene over-ex- injury model induced by adenovirus plasmid. Methods Seventy-two SD rats were randomly divided into normal group, non-transfection group, Ad-GFP transfection group and Ad-ANT1 transfection group （ 18 in each group）. ANT1 gene was trans- fected into rat carotid arteries with Ad-ANT1 adenovirus after a rat carotid balloon injury model was established. The arteries were harvested on days 7, 14 and 28 after operation. Expression of ANT-1, BAX, and Bcl-2 in arteries was detected by RT-PCR, Western blotting, and immunohistochemistry, respectively, with HE stai- ning. Apoptosis of VSMC in tunica intima and tunica media of the model was assayed with TUNEL staining. Results The ANT1 gene was significantly expressed in rat carotid arteries after Ad-ANT1 transfection and reached its peak level on day 14, which was significantly higher in Ad-ANT1 transfection group than in Ad-GFP transfection group and balloon injury group（P 〈 0.01 ）. The expression level of BAX was significantly higher in Ad-ANT1 transfection group than in Ad-GFP transfection group and balloon injury group on days 7 and 14 after transfection （P 〈 0. 05 ）. The expression level of Bcl-2 was higher in Ad-ANT1 transfection group than in normal group （P 〈 0. 05 ）. However, no significant difference was found in expression level of Bcl-2 between the two groups （ P 〉 0. 05 ）. The apoptosis rate of VSMC in tunica intima and tunica media of the model was significantly higher in Ad-ANT1 transfection group than in the other 3 groups （P 〈 0. 05 ）. The tunica intima/tunica mediaarea ratio in the model was lower in Ad-ANT1 transfection group than in Ad-GFP transfection group and balloon injury group on days 14 and 28 after transfection（P 〈 0. 05）. Gonclusion Adenovirus plasmid-induced over- expression of ANT1 gene induces apoptosis of VSMC in tunica inti

关 键 词：腺嘌呤核苷酸转位酶-1 颈动脉球囊损伤 血管平滑肌细胞 凋亡 

分 类 号：R543.402[医药卫生—心血管疾病]