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KMID : 0811720070110020045
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Korean Journal of Physiology & Pharmacology
2007 Volume.11 No. 2 p.45 ~ p.55
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Effects of pH, Buffer System and Lactate on the Simulated Ischemia-reperfusion Injury of H9c2 Cardiac Myocytes
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Lee Jun-Whee
Lee Hye-Kyung
Kim Hae-Won
Kim Young-Hoon
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Abstract
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We elucidated the effects of various components of ischemic medium on the outcome of simulated ischemia-reperfusion injury. Hypoxia for up to 12 hours induced neither apoptotic bodies nor LDH release. However, reoxygenation after 6 or 12 hours of hypoxia resulted in a marked LDH release along with morphological changes compatible with oncotic cell death. H9c2 cells were then subjected to 6 hours of simulated ischemia by exposing them to modified hypoxic glucose-free Krebs-Henseleit buffer. Lowered pH (pH 6.4) of simulated-ischemic buffer resulted in the generation of apoptotic bodies during ischemia, with no concomitant LDH release. The degree of reperfusion-induced LDH release was not affected by the pH of ischemic buffer. Removal of sodium bicarbonate from the simulated ischemic buffer markedly increased cellular damages during both the simulated ischemia and reperfusion. Addition of lactate to the simulated ischemic buffer increased apoptotic cell death during the simulated ischemia. Most importantly, concomitant acidosis and high lactate concentration in ischemic buffer augmented the reperfusion-induced oncotic cell death. These results confirmed the influences of acidosis, bicarbonate deprivation and lactate on the progression and outcome of the simulated ischemia-reperfusion, and also demonstrated that concomitant acidosis and high lactate concentration in simulated ischemic buffer contribute to the development of reperfusion injury.
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KEYWORD
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H9c2 cell, Simulated ischemia-reperfusion, Serum deprivation, Acidosis, Lactate, Apoptosis, Oncosis
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