Angiotensin II enhancement of hormone-stimulated cAMP formation in cultured vascular smooth muscle cells.
The mechanism by which angiotensin II (ANG II) potentiates hormone-induced adenosine 3',5'-cyclic monophosphate (cAMP) formation was studied in cultured rat vascular smooth muscle cells. Incubation of cells for 60 s with 100 nM ANG II produced a two- to threefold enhancement of cAMP stimulation when coupled with isoproterenol, prostaglandin I2, or adenosine. ANG II also enhanced cAMP formation when adenylyl cyclase was stimulated directly with forskolin or activated through the stimulatory guanyl nucleotide-binding protein (Gs) with cholera toxin. Forskolin stimulation was increased by only 40%, but cholera toxin-stimulated cAMP formation was doubled. Activation of protein kinase C with phorbol 12-myristate 13-acetate (PMA) enhanced isoproterenol-stimulated cAMP by 51%, but inhibitors of protein kinase activation had little effect on ANG II enhancement of cAMP production. However, use of PMA to cause feedback inhibition of D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] formation blocked the effect of ANG II on agonist-stimulated cAMP formation, and the time course for this effect of PMA paralleled its inhibitory effect on Ins(1,4,5)P3 production. Furthermore, chelation of intracellular Ca2+ or treatment with calmodulin antagonists also diminished the synergism between ANG II and isoproterenol for cAMP stimulation. The results indicate that ANG II enhances cAMP formation in vascular smooth muscle cells by facilitating the interaction between activated Gs and adenylyl cyclase. In addition, the data suggest that this effect of ANG II is directly related to Ins(1,4,5)P3 stimulation and appears to involve a Ca(2+)-calmodulin-dependent mechanism.
Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Charleston 29425.
S W
SW
Kubalak
J G
JG
Webb
D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis*; D004357:Drug Synergism; D006728:Hormones / Q000494:pharmacology*; D015544:Inositol 1,4,5-Trisphosphate / Q000502:physiology; D007425:Intracellular Membranes / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D006728:Hormones; D000804:Angiotensin II; D015544:Inositol 1,4,5-Trisphosphate; D002772:Cholera Toxin; D000242:Cyclic AMP; D011493:Protein Kinase C; D007545:Isoproterenol; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium