Involvement of Protein Kinase C and RhoA in Protease-Activated Receptor 1–Mediated F-Actin Reorganization and Cell Growth in Rat Cardiomyocytes
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概要
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Protease-activated receptor 1 (PAR1) that can be activated by serine proteinases such as thrombin has been demonstrated to contribute to the development of cardiac remodeling and hypertrophy after myocardial injury. Here, we investigated the mechanisms by which PAR1 leads to hypertrophic cardiomyocyte growth using cultured rat neonatal ventricular myocytes. PAR1 stimulation with thrombin (1 U/ml) or a synthetic agonist peptide (TFLLR-NH2, 50 μM) for 48 h induced an increase in cell size and myofibril formation associated with BNP (brain natriuretic peptide) production. This actin reorganization assessed by fluorescein isothiocyanate (FITC)-conjugated phalloidin staining appeared at 1 h after PAR1 stimulation, and this response was reduced by a protein kinase C (PKC) inhibitor, chelerythrine, inhibitors of Rho (simvastatin) and Rho-associated kinase (ROCK) (Y-27632), but not by pertussis toxin (PTX). By Western blot analysis, translocation of PKCα or PKCε from the cytosol to membrane fractions was observed in cells stimulated with thrombin or TFLLR-NH2 for 2 – 5 min. In addition, PAR1 stimulation for 3 – 5 min increased the level of active RhoA. Furthermore, inhibitors of PKC and ROCK and Rho abrogated PAR1-mediated increase in cell size. Depletion of PKCα or PKCε by specific small interfering RNA also suppressed both actin reorganization and cell growth. These results suggest that PAR1 stimulation of cardiomyocytes induces cell hypertrophy with actin cytoskeletal reorganization through activation of PKCα and PKCε isoforms and RhoA via PTX-insensitive G proteins.
著者
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NAKAMURA TOMOYUKI
Department of Orthopedic Surgery, Graduate School of Medical Sciences, Kyushu University
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Otani Hitomi
Department of Pharmacology, Kansai Medical University, Japan
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Yoshioka Kei
Department of Pharmacology, Kansai Medical University, Japan
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Nishikawa Hiroyuki
Fuso Pharmaceutical Industries, Ltd., Reseach & Development Center, Japan
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Inagaki Chiyoko
Department of Pharmacology, Kansai Medical University, Japan
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Otani Hitomi
Department Of Pharmacology Kansai Medical University
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Inagaki Chiyoko
Department Of Biology Kyoto College Of Pharmacy
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Nakamura Tomoyuki
Department Of Applied Chemistry Fuculty Of Engineering Naogya University
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Yoshioka Kei
Department Of Pharmacology Kansai Medical University
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