Novel and effective gene transfer technique for study of vascular renin angiotensin system.
Novel and effective gene transfer technique for study of vascular renin angiotensin system.
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用于研究血管肾素血管紧张素系统的新颖有效的基因转移技术。
DOI：
10.1172/jci116496
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Dzau,VJ
中科院分区：
文献类型：
--
作者：
Morishita,R;Gibbons,GH;Kaneda,Y;Ogihara,T;Dzau,VJ
血管肾素血管紧张素系统（RAS）存在于血管壁。然而，没有直接证据表明血管RAS本身是否可以调节血管平滑肌细胞（VSMC）的生长，因为没有合适的方法来研究内源性产生的血管活性物质对这些细胞生长的影响。在这项研究中，我们转移血管紧张素转换酶（ACE）和/或肾素cDNA到培养的VSMC使用有效的仙台病毒（日本血凝病毒）脂质体介导的基因转移方法，以检查它们在VSMC体外生长的相对作用。在35 min或6 h内，用日本血凝病毒法将ACE cDNA转染入VSMC，其ACE活性是对照载体的2倍，而阳离子脂质体（Lipofectin）介导的方法则没有显示出任何效果。这个体外系统为我们提供了研究内源性血管RAS对VSMC生长的影响的机会。ACE或肾素cDNA的转染导致DNA和RNA合成增加，其被特异性血管紧张素II受体拮抗剂（DuP 753：10（-6）M）抑制。血管紧张素I添加到ACE转染的VSMC中以剂量依赖性方式增加RNA合成。与单独的ACE或肾素cDNA相比，共转染的肾素和ACE cDNA刺激进一步的RNA合成。这些结果表明，RAS的转染组件可以调节VSMC的生长，通过血管紧张素II的内源性生产，以及ACE和肾素是速率限制在确定VSMC RAS活性。我们的结论是，日本血凝病毒脂质体介导的基因转移技术为研究内源性血管调节因子如血管RAS提供了一种新的有用工具。
Vascular renin angiotensin system (RAS) has been reported to exist in vascular wall. However, there is no direct evidence whether the vascular RAS per se can modulate growth of vascular smooth muscle cells (VSMC), because there is no suitable method to investigate the effect of endogenously produced vasoactive substances on growth of these cells. In this study, we transferred angiotensin-converting enzyme (ACE) and/or renin cDNAs into cultured VSMC using the efficient Sendai virus (hemagglutinating virus of Japan) liposome-mediated gene transfer method, to examine their relative roles in VSMC growth in vitro. Within 35 min or 6 h, the transfection of ACE cDNA into VSMC by hemagglutinating virus of Japan method resulted in a twofold higher ACE activity than control vector, whereas a cationic liposome (Lipofectin)-mediated method failed to show any effect. This in vitro system provided us with the opportunity to investigate the influence of endogenous vascular RAS on VSMC growth. Transfection of ACE or renin cDNA resulted in increased DNA and RNA synthesis, which was inhibited with the specific angiotensin II receptor antagonist (DuP 753: 10(-6) M). Angiotensin I added to ACE-transfected VSMC increased RNA synthesis in a dose-dependent manner. Cotransfection of renin and ACE cDNAs stimulated further RNA synthesis as compared to ACE or renin cDNA alone. These results showed that transfected components of RAS can modulate VSMC growth through the endogenous production of vascular angiotensin II, and that ACE as well as renin are rate limiting in determining the VSMC RAS activity. We conclude that the hemagglutinating virus of Japan liposome-mediated gene transfer technique provides a new and useful tool for study of endogenous vascular modulators such as vascular RAS.Images