Gene transfer of recombinant endothelial nitric oxide synthase to liver in vivo and in vitro.
Gene transfer of recombinant endothelial nitric oxide synthase to liver in vivo and in vitro.
复制标题
DOI：
10.1152/ajpgi.2000.279.5.g1023
复制
发表时间：
2000-11
期刊：
影响因子：
--
通讯作者：
V. Shah;Alex Chen;S. Cao;H. Hendrickson;D. Weiler;Leslie A. Smith;J. Yao;Z. Katusic
中科院分区：
文献类型：
--
作者：
V. Shah;Alex Chen;S. Cao;H. Hendrickson;D. Weiler;Leslie A. Smith;J. Yao;Z. Katusic
内皮型一氧化氮合酶（eNOS）衍生的一氧化氮（NO）有助于肝血管稳态。本研究的目的是研究是否递送编码eNOS基因的腺病毒载体到肝脏影响体内血管功能和体外NO产生的机制。通过尾静脉注射给予大鼠编码β-半乳糖苷酶（AdCMVLacZ）或eNOS（AdCMVeNOS）的腺病毒，并在1周后进行研究。在用AdCMVLacZ转导的动物中，β-半乳糖苷酶活性在肝脏中增加，在肝细胞中最显著。在AdCMVeNOS转导的动物中，eNOS蛋白水平和催化活性显著增加。eNOS的过度表达降低了灌注肝脏中对α（1）-激动剂甲氧胺的基线灌注压和收缩反应。用AdCMVeNOS转导培养的肝细胞导致重组eNOS靶向核周分布并与NOS激活蛋白热休克蛋白90结合。这些事件与离子霉素刺激的NO释放增加有关。总之，这是第一个研究，证明成功交付的重组eNOS基因在体内和体外肝脏，随后NO的生产。
Endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) contributes to hepatic vascular homeostasis. The aim of this study was to examine whether delivery of an adenoviral vector encoding eNOS gene to liver affects vasomotor function in vivo and the mechanism of NO production in vitro. Rats were administered adenoviruses encoding beta-galactosidase (AdCMVLacZ) or eNOS (AdCMVeNOS) via tail vein injection and studied 1 wk later. In animals transduced with AdCMVLacZ, beta-galactosidase activity was increased in the liver, most prominently in hepatocytes. In AdCMVeNOS-transduced animals, eNOS protein levels and catalytic activity were significantly increased. Overexpression of eNOS diminished baseline perfusion pressure and constriction in response to the alpha(1)-agonist methoxamine in the perfused liver. Transduction of cultured hepatocytes with AdCMVeNOS resulted in the targeting of recombinant eNOS to a perinuclear distribution and binding with the NOS-activating protein heat shock protein 90. These events were associated with increased ionomycin-stimulated NO release. In summary, this is the first study to demonstrate successful delivery of the recombinant eNOS gene to liver in vivo and in vitro with ensuing NO production.