title: Ca(2+) influx in the endothelial cells is required for the bradykinin-induced endothelium-dependent contraction in the porcine interlobar renal artery.
abstract: 1. To determine the mechanism of bradykinin-induced production of endothelium-derived contracting factors, we monitored the changes in cytosolic Ca(2+) concentration ([Ca(2+)](i)) in in situ endothelial cells in porcine aortic valvular strips and the changes in [Ca(2+)](i) of smooth muscle cells and force in porcine interlobar renal arterial strips using front-surface fluorometry of fura-2. 2. In the presence of N(omega)-nitro-L-arginine methyl ester, bradykinin caused an endothelium-dependent transient elevation of [Ca(2+)](i) and contraction in smooth muscle in the interlobar renal artery. This contraction was completely inhibited by a prostaglandin H(2)/thromboxane A(2) receptor antagonist. 3. In the absence of extracellular Ca(2+), bradykinin failed to induce contraction. However, replenishing extracellular Ca(2+) to 0.75 mM and higher induced an instantaneous contraction. However, replenishing Ca(2+) per se did not induce any contraction in the absence of bradykinin. Pretreatment with either 10(-5) M 1-(beta-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenethyl)-1H-imidazole hydrochloride (SKF96365) or 0.2 mM Ni(2+) abolished the contraction induced by bradykinin in the presence of extracellular Ca(2+). 4. Treatment with 10(-5) M indomethacin completely inhibited the contractile response induced by Ca(2+) replenishment, regardless of the timing of its application, before or after the application of bradykinin. 5. In endothelial cells in the valvular strips, bradykinin caused a transient [Ca(2+)](i) elevation in the presence of 1.25 mM extracellular Ca(2+), but [Ca(2+)](i) returned to the resting level within 10 min. Neither 10(-5) M SKF96365 nor 0.2 mM Ni(2+) had any effect on the peak [Ca(2+)](i) elevation, but decreased [Ca(2+)](i) in the declining phase. In the absence of extracellular Ca(2+), bradykinin induced a transient [Ca(2+)](i) elevation to a level similar to that seen in the presence of 1.25 mM extracellular Ca(2+). However, [Ca(2+)](i) then rapidly returned to the prestimulation level within 5 min. Subsequent Ca(2+) replenishment to 0.75 mM and higher in the presence of bradykinin elevated [Ca(2+)](i) to significantly higher levels than the resting level seen in the media containing 1.25 mM Ca(2+). 6. In conclusion, Ca(2+) influx in the endothelial cells is essential for bradykinin to induce endothelium-dependent contraction in the porcine interlobar renal artery.
authors.affiliation: Division of Molecular Cardiology, Research Institute of Angiocardiology, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Ihara
authors.forename: D N
authors.initials: DN
authors.lastname: Derkach
authors.forename: K
authors.initials: K
authors.lastname: Hirano
authors.forename: J
authors.initials: J
authors.lastname: Nishimura
authors.forename: H
authors.initials: H
authors.lastname: Nawata
authors.forename: H
authors.initials: H
authors.lastname: Kanaide
mesh_terms: D000818:Animals; D001920:Bradykinin / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology*; D005110:Extracellular Space / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D066298:In Vitro Techniques; D007213:Indomethacin / Q000494:pharmacology; D009532:Nickel / Q000494:pharmacology; D012077:Renal Artery / Q000187:drug effects / Q000502:physiology*; D013552:Swine; D014661:Vasoconstriction / Q000502:physiology*
chemical_list: D002121:Calcium Channel Blockers; D016861:Cyclooxygenase Inhibitors; D007093:Imidazoles; C022838:nickel chloride; D009532:Nickel; C063159:1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole; D001920:Bradykinin; D002118:Calcium; D007213:Indomethacin