Inhibitory Effect of Forskolin on Myosin Phosphorylation-Dependent and Independent Contractions in Bovine Tracheal Smooth Muscle.
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概要
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In bovine tracheal smooth muscle, carbachol (CCh, 1μM) and high K<SUP>+</SUP> (72.7mM) induced sustained increases in cytosolic Ca<SUP>2+</SUP>level ([Ca<SUP>2+</SUP>] i), myosin light chain (MLC) phosphorylation and force of contraction. Forskolin (FK, 1-10μM) inhibited the CCh-induced increase in [Ca<SUP>2+</SUP>MLC phosphorylation and force in parallel. In contrast, FK inhibited the high K<SUP>+</SUP>-induced contraction and MLC phosphorylation without changing [Ca<SUP>2+</SUP>]In the absence of extracellular Ca<SUP>2+</SUP> (with 0.5mM EGTA), CCh (10μM) and caffeine (20mM) induced transient increase in [Ca2+] i and contractile force by releasing Ca<SUP>2+</SUP>from cellular store. FK strongly inhibited the CCh-induced Ca<SUP>2+</SUP>transient, but failed to inhibit the caffeine-induced Ca2+transient. In the absence of external Ca<SUP>2+</SUP>, 12-deoxyphorbol 13-isobutylate (DPB, 1μM) induced sustained contraction without increase in [Ca<SUP>2+</SUP>] and MLC phosphorylation. FK inhibited this contraction without changing [Ca<SUP>2+</SUP>] In permeabilized muscle, Ca<SUP>2+</SUP>induced contraction in a concentration-dependent manner. FK (10μM) and cAMP (1-100μM) shifted the Ca<SUP>2+</SUP>-force curve to the higher Ca<SUP>2+</SUP>levels. CCh with GTP, GTPyS or DPB enhanced contraction in the presence of constant level of Ca<SUP>2+</SUP>. Forskolin and cAMP also inhibited the enhanced contractions in the permeabilized muscle. In the permeabilized, thiophosphorylated muscle, ATP induced contraction in the absence of Ca<SUP>2+</SUP>. cAMP (300μM) had no effect on this contraction. These results suggest that forskolin inhibits agonist-induced contraction in tracheal smooth muscle by multiple mechanisms of action; 1) inhibition of MLC phosphorylation by reducing Ca<SUP>2+</SUP>influx and Ca<SUP>2+</SUP>release, 2) inhibition of MLC phosphorylation by changing the MLC kinase/phosphatase balance, and 3) inhibition of regulatory mechanism which is not dependent on MLC phosphorylation.
著者
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Hori Masatoshi
Department Of Radiology Graduate School Of Medicine Osaka University
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Mitsui Minori
Department Of Veterinary Pharmacology Faculty Of Agriculture The University Of Tokyo
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TAJIMI Masaomi
Department of Veterinary Pharmacology, Graduate School of Agriculture and Life Sciences, The Univers
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KARAKI Hideaki
Life Sciences, The University of Tokyo
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OZAKI Hiroshi
Life Sciences, The University of Tokyo
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TAJIMI Masaomi
Department of Veterinary Pharmacology, Graduate School of Agriculture
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Tajimi Masaomi
Department of Veterinary Pharmacology, Faculty of Agriculture, The University of Tokyo
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HORI Masatoshi
Department of Veterinary Pharmacology, Graduate School of Agriculture
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MITSUI Minori
Department of Veterinary Pharmacology, Graduate School of Agriculture
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