Histamine-induced calcium transients in vascular smooth muscle cells: effects of verapamil and diltiazem.
We investigated the effects of verapamil and diltiazem on histamine-induced Ca2+ transients in vascular smooth muscle. 1) With the use of quin2 microfluorometry, cytosolic Ca2+ concentrations were directly measured in cultured vascular smooth muscle cells of the rat aorta. In the presence of extracellular Ca2+, histamine induced an elevation of cytosolic Ca2+ concentrations of a peak and plateau type. The peak component is due to a release of Ca2+ from cellular store sites, and the plateau component depends on extracellular Ca2+. Verapamil and diltiazem inhibited the plateau component, and the 50% inhibitive concentration (IC50) of verapamil and diltiazem for 10 microM histamine was 0.09 and 0.18 microM, respectively. Only at high concentrations did verapamil (IC50 = 8.7 microM) and diltiazem (IC50 = 95.7 microM) inhibit the Ca2+ release from the cellular store sites, as induced by 10 microM histamine. 2) Histamine, verapamil, and diltiazem competed with [3H]mepyramine for binding to the porcine aortic membranes, the order of potency being verapamil (Ki = 7.1 microM) greater than histamine (Ki = 18 microM) greater than diltiazem (Ki = 114 microM). From these results, we conclude that verapamil and diltiazem strongly inhibit the histamine-mediated, extracellular Ca2+-dependent intracellular [Ca2+] increase. In addition, verapamil and diltiazem seem to inhibit the release of Ca2+ from intracellular store sites, only at high concentrations, and probably by competing with histamine for binding to the H1-receptor. The inhibitory effects of Ca2+ antagonists on the histamine-induced contraction or spasm of vascular smooth muscle may well relate to these mechanisms.
Matsumoto
T
T
Division of Molecular Cardiology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Kanaide
H
H
Nishimura
J
J
Kuga
T
T
Kobayashi
S
S
Nakamura
M
M
D000634:Aminoquinolines; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002458:Cell Fractionation / Q000379:methods; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient / Q000379:methods; D002927:Cimetidine / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D004110:Diltiazem / Q000494:pharmacology*; D005456:Fluorescent Dyes; D006632:Histamine / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D008856:Microscopy, Fluorescence; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D011738:Pyrilamine / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012508:Sarcolemma / Q000378:metabolism / Q000648:ultrastructure; D013552:Swine; D014700:Verapamil / Q000494:pharmacology*
D000634:Aminoquinolines; D005456:Fluorescent Dyes; D002927:Cimetidine; D006632:Histamine; D014700:Verapamil; D004110:Diltiazem; D011738:Pyrilamine; C035739:Quin2; D002118:Calcium