Adenovirus-mediated transfer of human placental ectonucleoside triphosphate diphosphohydrolase to vascular smooth muscle cells suppresses platelet aggregation in vitro and arterial thrombus formation in vivo
Adenovirus-mediated transfer of human placental ectonucleoside triphosphate diphosphohydrolase to vascular smooth muscle cells suppresses platelet aggregation in vitro and arterial thrombus formation in vivo
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DOI：
10.1161/01.cir.0000155239.46511.79
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发表时间：
2005-02-15
期刊：
影响因子：
37.8
通讯作者：
Asada, Y
中科院分区：
文献类型：
--
作者：
Furukoji, E;Matsumoto, M;Asada, Y
背景-富血小板血栓形成是心血管疾病发病的关键事件。由于ADP在血小板聚集中起重要作用，因此其代谢在血小板活化和募集的调节中是重要的。外核苷三磷酸二磷酸水解酶（E-NTPDase）是参与血管ADP代谢的关键酶。我们最近从人胎盘中分离出两种E-NTPD酶亚型。本研究探讨这些亚型是否抑制血小板聚集和血栓形成后，腺病毒介导的基因转移到血管平滑肌细胞（SMCs）。方法和结果-我们构建腺病毒载体表达人胎盘E-NTPD酶亚型I（AdPlac I）和II（AdPlac II）或细菌β-半乳糖苷酶（AdLacZ）。AdPlac I感染的血管平滑肌细胞表达显著的NTPD酶活性，并抑制ADP和胶原诱导的血小板聚集。相反，用AdPlac II和AdLacZ感染的SMCs没有发挥抗血小板作用。为了研究胎盘E-NTPD酶亚型I在体内的抗血栓形成和抗增殖作用，我们在基因转移后5天通过全身给予虎红和经腔绿色光在大鼠颈动脉中产生血栓形成，并在血栓形成后3周检查新生内膜生长。AdLacZ感染动脉的血流迅速恶化，并在闭塞性血栓形成的96 +/-18秒内消失。相比之下，AdPlac I感染的动脉中的血流在照射期间被保存至少10分钟。此外，血栓形成和随后的新生内膜growth.Conclusions-胎盘E-NTPDase在损伤动脉的局部表达可能会防止动脉血栓形成和随后的新生内膜生长明显抑制。
Background - Platelet-rich thrombus formation is a critical event in the onset of cardiovascular disease. Because ADP plays a significant role in platelet aggregation, its metabolism is important in the regulation of platelet activation and recruitment. Ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) is a key enzyme involved in vascular ADP metabolism. We recently isolated 2 isoforms of E-NTPDase from the human placenta. The present study examined whether these isoforms suppress platelet aggregation and thrombus formation after adenovirus-mediated gene transfer to vascular smooth muscle cells (SMCs).Methods and Results - We constructed adenovirus vectors expressing human placental E-NTPDase isoforms I (AdPlac I) and II (AdPlac II) or bacterial beta-galactosidase (AdLacZ). Vascular SMCs infected with AdPlac I expressed significant NTPDase activity and inhibited the platelet aggregation induced by ADP and collagen in vitro. In contrast, SMCs infected with AdPlac II and AdLacZ did not exert antiplatelet effects. To investigate the antithrombotic and antiproliferative effects of placental E-NTPDase isoform I in vivo, we generated thrombosis in rat carotid arteries by systemically administered rose Bengal and transluminal green light 5 days after gene transfer and examined neointimal growth 3 weeks after thrombus formation. Blood flow in AdLacZ-infected arteries rapidly deteriorated and vanished within 96 +/- 18 seconds of occlusive thrombus formation. In contrast, blood flow in AdPlac I - infected arteries was preserved for at least 10 minutes during irradiation. In addition, thrombus formation and subsequent neointimal growth were obviously suppressed.Conclusions - The local expression of placental E-NTPDase in injured arteries might prevent arterial thrombosis and subsequent neointimal growth.