Effects of a Calcium-Channel Blocker (CV159) on Hepatic Ischemia/Reperfusion Injury in Rats: Evaluation with Selective NO/pO2 Electrodes and an Electron Paramagnetic Resonance Spin-Trapping Method
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概要
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Nitric oxide (NO) and the partial pressure of oxygen (pO2) in the liver were simultaneously quantified in rats with partial hepatic ischemia/reperfusion injury (PHIRI). Real-time NO/pO2 monitoring and immunohistochemical analysis for superoxide dismutase and inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) were performed to evaluate the protective effects of a dihydropyridine-type calcium-channel blocker—CV159—on PHIRI. Serum high-mobility-group box-1 (HMGB-1) was measured to assess cellular necrosis. Moreover, we used in vitro/ex vivo electron paramagnetic resonance spin trapping to assess the hydroxyl radical (·OH)-scavenging activity (OHSA) of CV159 and the liver tissue. The NO levels were significantly higher in CV159-treated rats than in control rats throughout the ischemic phase. Immediately after reperfusion, the levels temporarily increased in waves and then gradually decreased in the treated rats but remained constant in the control rats. pO2 was continually higher in the treated rats. In these rats, hepatic eNOS expression increased, whereas iNOS expression decreased. The treated rats exhibited significantly higher cytosolic and mitochondrial concentrations NOx (NO2+NO3). The serum HMGB-1 levels significantly decreased in the treated rats. Moreover, CV159 directly scavenged ·OH and both mitochondrial and cytosolic OHSA were preserved in the treated rats. Thus, CV159-mediated inhibition of intracellular Ca2+ overloading may effectively minimize organ damage and also have ·OH-scavenging activity and the cytoprotective effects of eNOS-derived NO.
著者
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KAMIBAYASHI Masato
Kyoto Pharmaceutical University
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Kawakami Hiroyoshi
Department Of Applied Chemistry Graduate Course Of Engineering Tokyo Metropolitan University
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Nagaya Masaki
Department Of Emergency And Critical Care Medicine St. Marianna University School Of Medicine
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Kobayashi Shinichi
Department Of Biomedical Engineering Graduate School Of Medicine University Of Tokyo
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Nakano Hiroshi
Department Of Anesthesia Okazaki City Hospital
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Kumai Toshio
Department of Pharmacogenomics, St. Marianna University School of Medicine
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Otsubo Takehito
Department of Gastroenterological and General Surgery, St. Marianna University Hospital
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Oowada Shigeru
Department of Gastroenterological and General Surgery, St. Marianna University Hospital
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Murakami Eiichi
Eikokagaku
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Kobayashi Shinichi
Department of Pharmacology, St. Marianna University Hospital
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Hataji Keizo
Department of Gastroenterological and General Surgery, St. Marianna University Hospital
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Watanabe Taiji
Hamamatsucho 1st Clinic Yokohama
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Ishiuchi Atsuko
Department of Gastroenterological and General Surgery, St. Marianna University Hospital
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