Perivascular gene transfer of NADPH oxidase inhibitor suppresses angioplasty-induced neointimal proliferation of rat carotid artery
Perivascular gene transfer of NADPH oxidase inhibitor suppresses angioplasty-induced neointimal proliferation of rat carotid artery
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DOI：
10.1152/ajpheart.00413.2004
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发表时间：
2005-02-01
影响因子：
4.8
通讯作者：
Pagano, PJ
中科院分区：
文献类型：
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作者：
Dourron, HM;Jacobson, GM;Pagano, PJ
血管牵张诱导NADPH氧化酶衍生的超氧阴离子（O-2（-）），这与肥大和细胞增殖有关。我们假设，靶向外膜递送NADPH氧化酶抑制剂将减少牵张诱导的血管O-2（-）并减弱新生内膜形成。我们设计了一种新的复制缺陷型腺病毒，它含有成纤维细胞活性启动子，驱动NADPH氧化酶抑制序列gp 91 ds的表达（Ad-PDGF betaR-gp 91 ds/eGFP）。1)我们使用pPDGF β R-荧光素酶通过在培养的大鼠主动脉成纤维细胞中显示荧光素酶的诱导而不是在血管平滑肌细胞中显示荧光素酶的诱导来表征该启动子的特异性。2)RT-PCR检测Ad-PDGF betaR-gp 91 ds/ eGFP感染成纤维细胞后gp 91 ds和报告基因的表达。3)使用Ad-CMV-eGFP作为对照，我们将Ad-PDGFbetaR-gp 91 ds/eGFP递送到大鼠颈总动脉（CCA）外膜。免疫组织化学证实了抑制剂局部递送到外膜。在用栓子切除导管损伤CCA后，我们观察到对照CCA中新生内膜与中膜面积比显著增加，而用表达gp 91 ds的病毒治疗的CCA中新生内膜与中膜面积比显著减弱。在第二组大鼠中，我们检测到膨胀刺激的O-2（-）增加了10倍，而在感染gp 91 ds表达病毒的CCA中，O-2（-）显著减少。这些数据表明，NADPH氧化酶抑制剂的局部外膜递送可有效减少总体血管O-2（-）和新生内膜形成，表明外膜NADPH氧化酶在新生内膜增生的发展中发挥功能性作用。
Vascular stretch induces NADPH oxidase-derived superoxide anion (O-2(-)), which has been implicated in hypertrophy and cell proliferation. We hypothesized that targeted delivery of an NADPH oxidase inhibitor to the adventitia would reduce stretch-induced vascular O-2(-) and attenuate neointima formation. We designed a novel replication-deficient adenovirus containing a fibroblast-active promoter driving expression of NADPH oxidase inhibitory sequence gp91ds (Ad-PDGF betaR-gp91ds/eGFP). 1) We characterized the specificity of this promoter using pPDGFbetaR-luciferase by showing induction of luciferase in cultured rat aortic fibroblasts but not in vascular smooth muscle cells. 2) Using RT-PCR, we observed expression of gp91ds and the reporter gene in fibroblasts after infection with Ad-PDGFbetaR-gp91ds/ eGFP. 3) Using Ad-CMV-eGFP as a control, we delivered Ad-PDGFbetaR-gp91ds/eGFP to the adventitia of the rat common carotid artery (CCA). Immunohistochemistry confirmed localized delivery of the inhibitor to the adventitia. After CCAs were injured with an embolectomy catheter, we observed a significant increase in neointima-to-media area ratio in control CCAs, which was significantly attenuated in CCAs treated with the gp91ds-expressing virus. In a second group of rats, we detected a 10-fold increase in distension-stimulated O-2(-), which was significantly reduced in CCAs infected with gp91ds-expressing virus. These data demonstrate that localized adventitial delivery of an NADPH oxidase inhibitor is effective in reducing overall vascular O-2(-) and neointima formation, suggesting that adventitial NADPH oxidase plays a functional role in development of neointimal hyperplasia.