Transfection of ACE or renin cDNA into cultured vascular smooth muscle cells using the HVJ liposome method resulted in a twofold increase in ACE activity and significantly increased DNA and RNA synthesis.
Does transfection of ACE and/or renin cDNAs using Sendai virus liposomes increase ACE activity and modulate growth in cultured vascular smooth muscle cells?
Sendai virus liposome-mediated gene transfer of ACE and renin cDNAs effectively demonstrates that endogenous vascular RAS components modulate VSMC growth.
p-value: p=<0.01
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.
Morishita et al. (Tue,) conducted a other in Vascular Renin Angiotensin System. HVJ liposome-mediated gene transfer of ACE and/or renin cDNAs vs. Control vector-transfected VSMC or Lipofectin-mediated transfection was evaluated on ACE activity and DNA/RNA synthesis (p=<0.01). Transfection of ACE or renin cDNA into cultured vascular smooth muscle cells using the HVJ liposome method resulted in a twofold increase in ACE activity and significantly increased DNA and RNA synthesis.