Ischemia depletes dystrophin and inhibits protein synthesis in the canine heart: mechanisms of myocardial ischemic injury.
In this study we tested our previous hypothesis that ischemia is a multifactorial injurious event involving all components of the myocyte simultaneously. This hypothesis was based on ultrastructural findings and was now tested again by protein analysis of sarcolemmal structural proteins and of markers of transcriptional and translational activities. This knowledge may help to clarify the cellular mechanisms involved in progression of acute ischemic myocardial injury and reperfusion. Therefore, we investigated all three intracellular/extracellular linkage systems of the sarcolemma using antibodies against dystrophin, beta-dystroglycan, gamma-sarcoglycan, vinculin, beta1-integrin, laminin, and spectrin. In addition, antibodies were used to evaluate membrane permeability (albumin), transcriptional efficacy (non-snRNP splicing factor SC-35), and translational capacity (phosphorylated p70 ribosomal protein S6 kinase). Tissue samples were obtained from a canine model of regional myocardial ischemia (90 min or 4.5 h) with or without reperfusion. Immunoconfocal microscopy and Western blotting revealed that the rank order of sensitivity was the following: dystrophin, beta-dystroglycan, gamma-sarcoglycan, vinculin, spectrin, integrin and laminin. Different levels of dystrophin loss indicate reversible/irreversible injury as established by albumin uptake and electron microscopy. Dystrophin depletion closely coincided with generally depressed transcription and translation. These changes occurred simultaneously in a time-dependent manner and persisted during reperfusion. In conclusion, damage of the different structural proteins results in membrane destabilization and disruption of the contractile apparatus from the sarcolemma. These changes, concomitantly associated with disturbances in transcription and translation, are major mechanisms determining the transition to irreversibility of myocardial ischemic injury and confirm our hypothesis that ischemia is a multifactorial injurious event involving all components of the cardiac myocyte.
Rodríguez
Manuel
M
Department of Experimental Cardiology, Max-Planck-Institute, Benekestr. 2, D-61231 Bad Nauheim, Germany.
Cai
Wei-Jun
WJ
Kostin
Sawa
S
Lucchesi
Benedict R
BR
Schaper
Jutta
J
D000418:Albumins / Q000378:metabolism; D000818:Animals; D004285:Dogs; D049030:Dystroglycans / Q000378:metabolism; D016189:Dystrophin / Q000378:metabolism*; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D017202:Myocardial Ischemia / Q000235:genetics / Q000378:metabolism*; D015428:Myocardial Reperfusion Injury / Q000235:genetics / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism; D014176:Protein Biosynthesis; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000378:metabolism; D049031:Sarcoglycans / Q000378:metabolism; D013049:Spectrin / Q000378:metabolism; D014158:Transcription, Genetic; D016596:Vinculin / Q000378:metabolism
D000418:Albumins; D016189:Dystrophin; D019012:Integrin beta1; D007797:Laminin; D049031:Sarcoglycans; D016596:Vinculin; D013049:Spectrin; D049030:Dystroglycans; D038762:Ribosomal Protein S6 Kinases, 70-kDa