BRADYKININ-B2 RECEPTOR EVOKED K+ PERMEABILITY INCREASE MEDIATES RELAXATION IN THE RAT DUODENUM

We have investigated the receptors and associated coupling mechanisms that mediate the smooth muscle relaxant response to bradykinin (BK) in the rat duodenum in vitro. Relaxation in response to BK seems due to a direct action on the longitudinal smooth muscle since effects were demonstrable in the presence of ibuprofen, mepyramine, atropine, guanethidine (all 1-mu-M), hexamethonium (10-mu-M) and TTX (0.3-mu-M). Receptors involved are of the B2 subtype since agonists and antagonists active at B1 receptors were essentially inactive, and the B2 receptor antagonist Lys,Lys-[Hyp3,Thi5,8,D-Phe7]BK was a potent competitive antagonist of BK-induced relaxation (pK(B) of 7.2 +/- 0.1). The activity of both BK and the antagonist were unchanged by the presence of peptidase inhibitors including the carboxypeptidase inhibitor DL-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid (mergetpa, 10-mu-M), which prevents conversion of BK analogues to des-Arg9-B1-active products. In high-K+ solution, BK (0.1-10-mu-M) produced concentration-related increases in Rb-86 efflux. Both this permeability increase in high-K+ solution, and the relaxant responses in Krebs solution, were inhibited by low concentrations (10-100 nM) of apamin, as well as the B2 receptor antagonist Lys,Lys-[Hyp3,Thi5,8,D-Phe7]BK (1-mu-M). These results are compatable with the proposal that BK-evoked relaxation of the rat duodenum is mediated via a subset of B2 receptors for which the antagonist Lys,Lys-[Hyp3,Thi5,8,D-Phe7]BK has a high affinity, and results from stabilisation of the smooth muscle membrane through the opening of apamin-sensitive Rb-86-permeable calcium-activated K+ channels.