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Am J Physiol Heart Circ Physiol 278: H367-H372, 2000;
0363-6135/00 $5.00
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Vol. 278, Issue 2, H367-H372, February 2000

B1 and B2 bradykinin receptors on adventitial fibroblasts of cerebral arteries are coupled to recombinant eNOS

Masato Tsutsui1, Hisashi Onoue1, Yasuhiko Iida1, Leslie Smith1, Timothy O'Brien2, and Zvonimir S. Katusic1

1 Departments of Anesthesiology and Pharmacology, and 2 Endocrinology and Metabolism, Mayo Clinic, Rochester, Minnesota 55905

Our previous ex vivo and in vivo studies reported that expression of the recombinant endothelial nitric oxide (NO) synthase (eNOS) gene in adventitial fibroblasts recovers NO production in arteries without endothelium in response to bradykinin. The present study was designed to characterize subtypes of bradykinin receptors on adventitial fibroblasts coupled to the activation of recombinant eNOS. Endothelium-denuded segments of canine basilar arteries were transduced with beta -galactosidase (beta -Gal) gene or eNOS gene ex vivo, using a replication-defective adenoviral vector (1010 plaque-forming units/ml) for 30 min at 37°C. Twenty-four hours later, isometric force recording or cGMP measurement was carried out. B1 bradykinin receptor agonist (des-Arg9-bradykinin, 10-10-10-8 mol/l) did not significantly affect vascular tone in control or beta -Gal gene-transduced canine basilar arteries without endothelium. In contrast, this agonist caused concentration-dependent relaxations in recombinant eNOS gene-transduced arteries without endothelium. Relaxations to B1 receptor agonist in the eNOS arteries were abolished by B1 receptor antagonist (des-Arg9-[Leu8]bradykinin, 6 × 10-9 mol/l) but not by B2 receptor antagonist (Hoe-140, 5 × 10-8 mol/l). Bradykinin did not significantly alter vascular tone in control or beta -gal arteries without endothelium, whereas this peptide (10-11-10-8 mol/l) induced concentration-dependent relaxations, as well as an increase in cGMP formation in endothelium-denuded eNOS-transduced arteries. Stimulatory effects of bradykinin were prevented in the presence of a B2 receptor antagonist but not in the presence of a B1 receptor antagonist. B1 and B2 receptor antagonists had no effect on relaxations to substance P, confirming the selectivity of the compounds. Our results suggest that B1 and B2 bradykinin receptors are coupled to activation of recombinant eNOS expressed in adventitial fibroblasts.

des-Arg9-[Leu8]bradykinin; Hoe-140; gene transfer


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