Novel and effective gene transfer technique for study of vascular renin angiotensin system.
Open Access
- 1 June 1993
- journal article
- research article
- Published by American Society for Clinical Investigation in JCI Insight
- Vol. 91 (6), 2580-2585
- https://doi.org/10.1172/jci116496
Abstract
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.Keywords
This publication has 18 references indexed in Scilit:
- Alpha 1-adrenoreceptor blockade reduces the angiotensin II-induced vascular smooth muscle cell DNA synthesis in the rat thoracic aorta and carotid artery.Circulation Research, 1992
- Induction of platelet-derived growth factor A-chain and c-myc gene expressions by angiotensin II in cultured rat vascular smooth muscle cells.JCI Insight, 1989
- Increased Expression of DNA Cointroduced with Nuclear Protein in Adult Rat LiverScience, 1989
- Molecular biology of the renin-angiotensin systemAmerican Journal of Physiology-Renal Physiology, 1988
- Angiotensin II induces hypertrophy, not hyperplasia, of cultured rat aortic smooth muscle cells.Circulation Research, 1988
- Vascular renin-angiotensin system in two-kidney, one clip hypertensive rats.Hypertension, 1986
- Angiotensinogen gene is expressed and differentially regulated in multiple tissues of the rat.JCI Insight, 1986
- Expression of smooth muscle-specific alpha-isoactin in cultured vascular smooth muscle cells: relationship between growth and cytodifferentiation.The Journal of cell biology, 1986
- Localization and Properties of Angiotensin ReceptorsJournal Of Hypertension, 1985
- Renin synthesis by canine aortic smooth muscle cells in cultureLife Sciences, 1982