Ischemic myocardial injury in cultured heart cells: Leakage of cytoplasmic enzymes from injured cells |
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Authors: | Daniel Acosta Marianne Puckett Reagan McMillin |
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Institution: | (1) Department of Pharmacology, College of Pharmacy, University of Texas, 78712 Austin, Texas |
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Abstract: | Summary An in vitro model of myocardial ischemia has been established with primary monolayer cultures of postnatal rat myocardial
cells. Ischemic conditions were simulated in vitro by subjecting the myocardial cell cultures to various levels of oxygen
and glucose deprivation. The experimental protocol consisted of treatment with 20% or 0% O2 and 1000, 500 or 0 mg glucose per 1 of medium for 4 or 24 hr. Control cultures were treated with 20% O2 and 1000 mg glucose. After the ischemic treatments, cultures of beating muscle (M) cells were evaluated for signs of injury,
i.e. leakage of cytoplasmic enzymes into the culture medium. Differences were found in leakage of lactate dehydrogenase (LDH)
and creatine phosphokinase (CPK) from the cultures that were exposed to partial ischemia of glucose deprivation and from those
cultures that were exposed to total ischemia of oxygen and glucose deprivation. Glucose deprivation alone resulted in a slight-to-moderate
loss of LDH and CPK from the cells, whereas total ischemia resulted in a significant release of the two cytoplasmic enzymes.
When the cultures were allowed to recover after ischemic treatment in complete medium (1000 mg glucose) and a normal atmosphere
of 20% O2, they had levels of LDH leakage comparable to those of control cultures. Cell viability and total protein content of the
ischemic cultures did not differ significantly from controls.
This study was supported by Research Grant HL 18647 from the National Heart, Lung, and Blood Institute. |
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Keywords: | myocardial ischemia cell injury lactate dehydrogenase creatine phosphokinase cultured myocardial cells glucose deprivation |
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