Calpain is Implicated in Rat Myocardial Injury after Ischemia or Reperfusion

元データ 1994-12-20 社団法人日本循環器学会

概要

Calpain activity was measured in the various subfractions of rat myocardia after global ischemia for 60 min or after ischemia followed by 30 min of reperfusion after the chromatographic separation of μ-and m-calpains. The activity of m-calpain after ischemia and that of μ-calpain after reperfusion were both higher than that in the control. The activity of the endogenous calpain inhibitor calpastatin in 10, 000×g supernatant was decreased after both ischemia and ischemia-reperfusion. The increase in m-and μ-calpain activities was suppressed by pre-ischemic perfusion with a synthetic calpain inhibitor, transepoxysuccinyl-L-leucylamido (4-guanidino) butane (E64d, 100 μg/ml). After reperfusion, the calpain activity in the 10, 000×g pellet was also increased, which was inhibited by pre-ischemic perfusion with E64d or dimethylsulfoxide (a solvent for E64d) or by reperfusion with 1 mmol/L ethyleneglycol bis (β-aminoethylether) -N, N, N', N'-tetraacetic acid. SDS-polyacrylamide gel electrophoresis revealed the proteolysis of several proteins, including fodrin, in the 10, 000×g and 100, 000×g pellet fractions after ischemia and reperfusion, some of which were confirmed to be in vitro substrates of calpain. The creatine kinase release during the reperfusion was also partially inhibited by E64d or dimethylsulfoxide. Thus, calpain activity in the soluble or particulate fractions was altered during ischemia or reperfusion, and appeared to be implicated in the proteolysis of the membrane proteins, which may contribute to myocardial injury.

著者

HIRONAKA Kouji Department of Gastroenterology and Hepatology, Yamaguchi University School of Medicine
Yoshida Ken-ichi Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University
Hironaka Kouji Department Of Gastroenterology And Hepatology Yamaguchi University School Of Medicine
Hironaka Kouji Department Of Legal Medicine Yamaguchi University School Of Medicine. Supported In Part By Grants Fr
Fujiwara Masao Department Of Plastic And Reconstructive Surgery Hamamatsu University School Of Medicine
Fujiwara Masao Department Of Legal Medicine Yamaguchi University School Of Medicine. Supported In Part By Grants Fr
SORIMACHI Yoshihide Department of Legal Medicine, Yamaguchi University School of Medicine
Yoshida Ken-ichi Department Of Legal Medicine Yamaguchi University School Of Medicine. Supported In Part By Grants Fr
Yoshida Ken-ichi Department Of Agrobioscience Graduate School Of Agricultural Science Kobe University
Sorimachi Yoshihide Department Of Legal Medicine Yamaguchi University School Of Medicine. Supported In Part By Grants Fr
Sorimachi Yoshihide Department Of Legal Medicine Kyoto Prefectural University Of Medicine
Fujiwara Masao Department Of Gastroenterological Surgery Faculty Of Medicine Kagawa University

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