Rho-kinase-mediated pathway induces enhanced myosin light chain phosphorylations in a swine model of coronary artery spasm
| Autor: | Kozo Kaibuchi, Akira Takeshita, Kouichi Kuwata, Kensuke Egashira, Naoki Katsumata, Toshiyuki Kozai, Minoru Seto, Hiroaki Shimokawa, Ichiro Ikegaki, Mutsuki Amano, Tadashi Kandabashi, T. Asano, Tohru Yamawaki |
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| Rok vydání: | 2000 |
| Předmět: |
Male
Serotonin Myosin light-chain kinase Myosin Light Chains Physiology Swine Blotting Western Coronary Vasospasm macromolecular substances Pharmacology In Vitro Techniques Protein Serine-Threonine Kinases Western blot In vivo Physiology (medical) 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine Myosin Medicine Animals Enzyme Inhibitors Phosphorylation Rho-associated protein kinase Protein kinase C Analysis of Variance rho-Associated Kinases medicine.diagnostic_test Kinase business.industry Intracellular Signaling Peptides and Proteins Anatomy Coronary Vessels Blot Vasodilation Cardiology and Cardiovascular Medicine business Interleukin-1 Signal Transduction |
| Zdroj: | Cardiovascular research. 43(4) |
| ISSN: | 0008-6363 |
| Popis: | Objective: We recently demonstrated in our swine model of coronary artery spasm that enhanced myosin light chain (MLC) phosphorylations (both MLC mono- and diphosphorylations) play a central role in the pathogenesis of the spasm. However, the molecular mechanism for and the phosphorylation sites for the enhanced MLC phosphorylations were unknown. In the present study, we addressed these points using hydroxyfasudil, a novel inhibitor of protein kinases, which we found preferentially inhibits Rho-kinase. Methods: The specificity of the inhibitory effects of hydroxyfasudil on Rho-kinase, MLCK, MRCKβ and PKC were examined by kinase assay in vitro. The left porcine coronary artery was chronically treated with interleukin-1β (IL-1β, 2.5 μg). Two weeks after the operation, coronary artery vasomotion was examined both in vivo and in vitro. MLC phosphorylations were examined by Western blot analysis and the sites for the phosphorylations by anti-phosphorylated MLC antibodies that identified the monophosphorylation site as Ser19 and diphophorylation sites as Ser19/Thr18 of MLC. Results: Inhibitory effects of hydroxyfasudil was at least 100 times more potent for Rho-kinase as compared with other protein kinases tested. Intracoronary serotonin (10 μg/kg) caused coronary hyperconstriction at the IL-1β-treated site in vivo, which was dose-dependently inhibited by hydroxyfasudil ( p |
| Databáze: | OpenAIRE |
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