腺病毒介导高血压相关基因转移对兔颈动脉球囊损伤后新生内膜形成的抑制作用

Adenovirus-mediated hypertension-related gene transfer to rabbit carotid ateries inhibits neointimal formation after balloon angioplasty

目的 探讨腺病毒载体介导的新基因高血压相关基因(Hypertension-related gene,HRG-1)转移对兔颈总动脉球囊损伤后新生内膜形成的影响。方法 兔颈总动脉球囊损伤后,经血管腔内转染含有HRG-1的腺病毒载体(AdHRG-1)或含有绿色荧光蛋白报告基因的空载腺病毒载体(AdGFP),以生理盐水处理为对照。于术后3、7和28 d取材,分别进行外源基因表达检测、PCNA染色和定量组织形态学分析。结果 应用逆转录-聚合酶链反应和免疫组织化学方法分别证实了基因转移后3 d HRG-1在兔颈动脉壁内的表达;PCNA免疫组织化学染色显示,转染AdHRG-1后7 d兔颈动脉内膜和中膜PCNA阳性细胞指数较转染AdGFP组明显降低[(21.4±2.5)％对(45.6±3.8)％,(6.4±1.1)％对(17.8±1.9)％,P<0.05]。基因转移后28 d血管壁定量组织形态学分析显示,转染AdHRG-1组血管新生内膜面积及内膜面积与中膜面积比均显著低于转染AdGFP组[(0.13±0.03)mm2对(0.45±0.14)mm2,0.19±0.02对0.70±0.15,P<0.01],血管腔面积则显著增加[(0.88±0.07)mm2对(0.57±0.27)mm2,P<0.01],而转染AdGFP组与生理盐水处理组间无显著性差异。结论 以腺病毒为载体经血管腔内转染HRG-1可有效抑制球囊损伤后血管平滑肌细胞增殖和新生内膜形成。

Objective To study the effects of adenovirus-mediated gene transfer of hypertension-related gene ( HRG-1) on neointimal formation in rabbit carotid arteries after balloon angioplasty. Methods The rabbit common carotid arteries subjected to balloon injury were treated with AdHRG-1, AdGFP or saline from lumen. The arteries were harvested 3 days after gene transfer for assessment of transgene expression, 7 days for immunostaining of proliferating cell nuclear antigen ( PCNA) and 28 days for quantitative histomorphometric analysis. Results Transgene expression in the vessel segment infected with AdHRG-1 was confirmed by RT-PCR and immunohistochemistry. The PCNA-positive index in the neointima and media at day 7 after transfection was significantly reduced in AdHRG-1 treated arteries than that in AdGFP treated arteries[ (21.4±2.5)% vs. ( 45. 6±3. 8)% ,(6. 4±1. 1)% vs. ( 17. 8±1. 9)% ,P < 0. 05 ]. The intimal area and the intimal/medial ratio ( I/M ) at day 28 were significantly reduced in the AdHRG-1 group than those in AdGFP group [ (0. 13± 0. 03) mm2 vs. ( 0.45 ± 0. 14) mm2,0. 19±0. 02 vs. 0.70 ±0. 15,P<0.01] ,the vessel lumen area was increased[ (0. 88±0.07) mm2 vs. (0.57±0.27) mm ,P <0. 01 ]. There were no significant differences between AdGFP group and saline group. Conclusion Adenovirus-mediated gene transfer of HRG-1 from lumen may effectively inhibit VSMC proliferation and neointimal formation in the rabbit carotid arteries after balloon angioplasty and may potentially be developed as a new approach to prevent restenosis after angioplasty.