Janssens S, Flaherty D, Nong Z, Varenne O, van Pelt N, Haustermans C, Zoldhelyi P, Gerard R, Collen D
Cardiac Unit, University Hospital Gasthuisberg, University of Leuven, Belgium.
Circulation. 1998 Apr 7;97(13):1274-81. doi: 10.1161/01.cir.97.13.1274.
Loss of endothelial NO production after arterial injury may contribute to restenosis, characterized by neointima formation and elastic recoil. Adenovirus-mediated transfer of the gene encoding NO synthase (NOS) in balloon-injured arteries may restore NO production and inhibit neointima formation.
After balloon injury, rat carotid arteries were transduced with 3x10(10) pfu/mL recombinant adenovirus carrying the human endothelial constitutive NOS cDNA (AdCMVceNOS, n=8) or no cDNA (AdRR5, n=8). ceNOS expression was confirmed by immunoblot analysis of vascular extracts and was localized by immunostaining in 30% of medial smooth muscle cells (SMCs) and in the adventitia of AdCMVceNOS-transduced arteries. Vascular cGMP levels were reduced from 3.9 pmol/g wet wt in uninjured arteries to 0.7 pmol cGMP/g after AdRR5 but were restored after ceNOS gene transfer (3.8 pmol cGMP/g wet wt, P<.05 versus AdRR5). Intima-to-media ratio 2 weeks after injury was significantly reduced (0.19+/-0.02 in AdCMVceNOS-infected versus 0.69+/-0.07 in AdRR5-infected arteries, P<.05). In vitro, BrdU incorporation of AdCMVceNOS-infected SMCs was reduced by 28% compared with AdRR5-infected SMCs. Transduced cells from injured carotid arteries subjected to FACS sorting showed a significantly lower BrdU labeling index in ceNOS-infected rats (29+/-6% versus 43+/-5% and 45+/-4% in control, injured, and AdRR5-infected rats, respectively, P<.05).
AdCMVceNOS gene transfer to balloon-injured rat carotid arteries restores vascular NO production and reduces neointima formation, at least in part because of an antiproliferative effect on medial SMCs. Adenovirus-mediated ceNOS gene transfer might reduce arterial restenosis after balloon angioplasty.
动脉损伤后内皮一氧化氮生成的丧失可能导致再狭窄，其特征为新生内膜形成和弹性回缩。腺病毒介导的一氧化氮合酶（NOS）编码基因在球囊损伤动脉中的转移可能恢复一氧化氮生成并抑制新生内膜形成。
球囊损伤后，用携带人内皮组成型 NOS cDNA 的 3×10(10) pfu/mL 重组腺病毒（AdCMVceNOS，n = 8）或无 cDNA 的腺病毒（AdRR5，n = 8）转导大鼠颈动脉。通过对血管提取物的免疫印迹分析证实了 ceNOS 的表达，并通过免疫染色在 30% 的中膜平滑肌细胞（SMC）和 AdCMVceNOS 转导动脉的外膜中定位。血管 cGMP 水平从未损伤动脉中的 3.9 pmol/g 湿重降至 AdRR5 处理后的 0.7 pmol cGMP/g，但在 ceNOS 基因转移后恢复（3.8 pmol cGMP/g 湿重，与 AdRR5 相比，P <.05）。损伤后 2 周的内膜与中膜比值显著降低（AdCMVceNOS 感染组为 0.19±0.02，AdRR5 感染组为 0.69±0.07，P <.05）。在体外，与 AdRR5 感染的 SMC 相比，AdCMVceNOS 感染的 SMC 的 BrdU 掺入减少了 28%。对损伤颈动脉进行 FACS 分选得到的转导细胞显示，ceNOS 感染大鼠的 BrdU 标记指数显著降低（分别为 29±6%，而对照组、损伤组和 AdRR5 感染组分别为 43±5% 和 45±4%，P <.05）。
AdCMVceNOS 基因转移至球囊损伤的大鼠颈动脉可恢复血管一氧化氮生成并减少新生内膜形成，至少部分原因是对中膜 SMC 具有抗增殖作用。腺病毒介导的 ceNOS 基因转移可能减少球囊血管成形术后的动脉再狭窄。
