title: Angiotensin IV receptor-mediated activation of lung endothelial NOS is associated with vasorelaxation.
abstract: The hexapeptide angiotensin (ANG) IV, a metabolic product of ANG II, has been reported to play a functional role in the regulation of blood flow in extrapulmonary tissues. Here, we demonstrate that ANG IV-specific (AT4) receptors are present in porcine pulmonary arterial endothelial cells (PAECs) and that the binding of ANG IV to AT4 receptors can be blocked by its antagonist divalinal ANG IV but not by the ANG II-, AT1-, and AT2-receptor blockers [Sar1,Ile8]ANG II, losartan, and PD-123177, respectively. ANG IV significantly increased endothelial cell constitutive nitric oxide synthase (ecNOS) activity (P < 0.05) as well as cellular cGMP content (P < 0. 001). Western blot analysis revealed that ecNOS protein expression was comparable in control and ANG IV-stimulated cells. Divalinal ANG IV but not [Sar1,Ile8]ANG II, losartan, or PD-123177 inhibited the ANG II- and ANG IV-stimulated increases in ecNOS activity and cGMP content in PAECs. Incubation in the presence of N-nitro-L-arginine methyl ester (L-NAME) or methylene blue but not of indomethacin significantly diminished ANG IV-stimulated as well as basal levels of cGMP (P < 0.001). Similarly, in situ studies with precontracted porcine pulmonary arterial rings showed that ANG IV caused an endothelium-dependent relaxation that was blocked by L-NAME or methylene blue. Collectively, these results demonstrate that ANG IV binds to AT4 receptors, activates ecNOS by posttranscriptional modulation, stimulates cGMP accumulation in PAECs, and causes pulmonary arterial vasodilation, suggesting that ANG IV plays a role in the regulation of blood flow in the pulmonary circulation.
authors.affiliation: Research Service, Department of Veterans Affairs Medical Center, University of Florida College of Medicine, Gainesville, Florida 32608-1611, USA.
authors.forename: J M
authors.initials: JM
authors.lastname: Patel
authors.forename: J R
authors.initials: JR
authors.lastname: Martens
authors.forename: Y D
authors.initials: YD
authors.lastname: Li
authors.forename: C H
authors.initials: CH
authors.lastname: Gelband
authors.forename: M K
authors.initials: MK
authors.lastname: Raizada
authors.forename: E R
authors.initials: ER
authors.lastname: Block
mesh_terms: D000626:Aminopeptidases / Q000378:metabolism; D000804:Angiotensin II / Q000031:analogs & derivatives / Q000494:pharmacology / Q000502:physiology; D057911:Angiotensin Receptor Antagonists; D000818:Animals; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D004789:Enzyme Activation / Q000502:physiology; D043384:Glutamyl Aminopeptidase; D007213:Indomethacin / Q000494:pharmacology; D063208:Methionyl Aminopeptidases; D008751:Methylene Blue / Q000494:pharmacology; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D011651:Pulmonary Artery / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D011945:Receptors, Angiotensin / Q000502:physiology*; D013552:Swine; D014664:Vasodilation / Q000502:physiology*
chemical_list: C090830:AT4 receptor; D057911:Angiotensin Receptor Antagonists; D011945:Receptors, Angiotensin; D000804:Angiotensin II; C012877:angiotensin II, des-Asp(1)-des-Arg(2)-Ile(5)-; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases; D043384:Glutamyl Aminopeptidase; D006152:Cyclic GMP; D008751:Methylene Blue; D019331:NG-Nitroarginine Methyl Ester; D007213:Indomethacin