Prevention of arterial thrombosis by adenovirus-mediated transfer of cyclooxygenase gene.
Prevention of arterial thrombosis by adenovirus-mediated transfer of cyclooxygenase gene.
复制标题
DOI：
10.1161/01.cir.93.1.10
复制
发表时间：
1996
期刊：
影响因子：
37.8
通讯作者：
P. Zoldhelyi;J. McNatt;Xiao-Ming Xu;D. Loose-Mitchell;R. S. Meidell;Fred J. Clubb;L. Buja;J. Willerson;J. Willerson;Kenneth K. Wu
中科院分区：
文献类型：
--
作者：
P. Zoldhelyi;J. McNatt;Xiao-Ming Xu;D. Loose-Mitchell;R. S. Meidell;Fred J. Clubb;L. Buja;J. Willerson;J. Willerson;Kenneth K. Wu
前列环素是一种重要的血管保护分子。它抑制血小板聚集、单核细胞与内皮细胞的相互作用和平滑肌细胞脂质积聚。血管环氧化酶-1（考克斯-1）是前列环素合成的限速步骤。本研究的目的是确定在猪颈动脉血管成形术模型中，腺病毒介导的考克斯-1转移是否可以恢复考克斯-1活性，增加前列环素合成，并防止血栓形成。方法与结果将人考克斯1 cDNA在巨细胞病毒启动子的驱动下，通过同源重组的方法构建到复制缺陷型腺病毒5型载体中。包括不含外源基因的重组腺病毒（Ad-RR）和缓冲液作为对照。重组Ad-LacZ用于体内标记转染细胞。在体外实验中，培养的人内皮细胞（EC）和猪动脉平滑肌细胞（SMC）与Ad-COX-1孵育2 h，并在感染后72 h测定6-keto-PGF（1 α）水平和转基因表达。在体内实验中，将重组腺病毒直接滴注到血管成形术损伤的猪颈动脉中30分钟。监测循环血流变化10天，此后对血栓形成进行组织学检查。测定损伤动脉的转基因表达和前列腺素I2（PGI 2）合成。用Ad-COX-1感染培养的内皮细胞，可使PGI 2表达增加5 ~ 8倍，北方分析证实了转基因在培养的猪平滑肌细胞中的表达。直接给予3 x 10（10）pfu剂量的Ad-COX-1完全抑制了颈动脉循环血流变化和血栓形成，并在感染后10天使受损动脉的PGI 2合成增加了四倍，而较低剂量的Ad-COX-1（5 x 10（10）pfu）与Ad-RR载体和缓冲液对照相比，没有抗血栓形成作用。结论：腺病毒介导的考克斯-1转移到血管成形术损伤的颈动脉是有效的，在增加PGI 2的合成，并与血栓形成的抑制，当滴注相对高滴度的腺病毒。
BACKGROUND Prostacyclin is an important vasoprotective molecule. It inhibits platelet aggregation, monocyte interaction with endothelium, and smooth muscle cell lipid accumulation. Vascular cyclooxygenase-1 (COX-1) is the rate-limiting step in prostacyclin synthesis. The objective of this study was to determine whether adenovirus-mediated transfer of COX-1 could restore COX-1 activity, augment prostacyclin synthesis, and prevent thrombus formation in a porcine carotid angioplasty model. METHODS AND RESULTS Human COX-1 cDNA driven by a cytomegalovirus promoter was constructed into a replication-defective adenovirus 5 vector by homologous recombination. Recombinant adenovirus without a foreign gene (Ad-RR) and buffer were included as controls. Recombinant Ad-LacZ was used for marking the transfected cells in vivo. In the in vitro experiments, cultured human endothelial cells (ECs) and porcine arterial smooth muscle cells (SMCs) were incubated with Ad-COX-1 for 2 hours and 6-keto-PGF(1 alpha) level and the transgene expression were determined 72 hours after infection. In the in vivo experiments, recombinant adenoviruses were directly instilled into angioplasty-injured porcine carotid arteries for 30 minutes. Cyclic flow changes were monitored for 10 days and thrombus formation was examined histologically thereafter. Transgene expression and prostaglandin I2 (PGI2) synthesis by the injured arteries were determined. Cultured ECs infected with Ad-COX-1 produced a fivefold to eightfold increase in PGI2, and the transgene expression in cultured porcine SMCs was demonstrated by Northern analysis. Direct administration of Ad-COX-1 at a dose of 3 x 10(10) pfu completely inhibited carotid cyclic flow changes and thrombus formation accompanied by a fourfold increase in PGI2 synthesis by the injured arteries 10 days after infection, whereas Ad-COX-1 at a lower dose, 5 x 10(9) pfu, had no antithrombotic effects when compared with Ad-RR vector and buffer controls. CONCLUSIONS Adenovirus-mediated transfer of COX-1 to angioplasty-injured carotid arteries was efficacious in augmenting PGI2 synthesis and was associated with an inhibition of thrombosis when a relatively high titer of adenovirus was instilled.