Ethanol hyperpolarizes mitochondrial membrane potential and increases mitochondrial fraction in cultured mouse myocardial cells.
Cultured mouse heart-derived myocardial and non-muscle cells were exposed to ethanol, stained with cell-permeant fluorescent vital probes, JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide) and oxidation-sensitive dihydrorhodamine 123, and analyzed by flow cytometry to elucidate ethanol-induced time-wise alterations in the mitochondrial membrane potential (DeltaPsim) and the production of reactive oxygen species (ROS). Ethanol (50 and 200 mM) not only hyperpolarized DeltaPsim of both types of cells but also dose-dependently increased ROS production at 24 h, although a 200-mM dose reduced the production until 3 h. These cell pathophysiological reactions suggest the depression of mitochondrial ATPase and mitochondrial respiratory chain. However, differences between these cells appeared after a 24-h exposure to 200 mM ethanol: the increase in ROS production was approximately twice as large for myocardial cells as for non-muscle cells; and the side-scatter parameter of light scattering significantly increased for myocardial cells, but not for non-muscle cells. All these myocyte-specific alterations indicate an increase in the mitochondrial fraction in a cell. This reaction might be a countermeasure against ethanol-induced dysfunction of mitochondrial respiration that is needed to meet the energy requirements of spontaneous myocardial contractions.
Department of Legal Medicine, Nippon Medical School, 1-1-5, Sendagi, 113-8602, Tokyo, Japan. kei-mam@nms.ac.jp
Keiko
K
Mashimo
Youkichi
Y
Ohno
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000633:toxicity*; D005434:Flow Cytometry; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000201:enzymology; D051336:Mitochondrial Membranes / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013997:Time Factors
D002492:Central Nervous System Depressants; D017382:Reactive Oxygen Species; D000431:Ethanol; D000251:Adenosine Triphosphatases