title: Phosphatidylcholine metabolism in isolated rat heart: modulation by ethanol and vitamin E.
abstract: Prolonged ethanol administration has been reported to cause defects in cardiac performance and abnormal cardiac lipid contents. However, little is known regarding the short-term administration of ethanol to the perfused heart and its effect on cardiac phospholipid metabolism. In this study, the isolated Langendorff heart perfusion was used as a model to study the effects of ethanol and a combination of ethanol and vitamin E (DL-alpha-tocopherol) on phospholipid metabolism. When perfused with 1% ethanol for 4 h, the major cardiac phospholipids were not altered but a 60% increase in lysophosphatidylcholine level was observed. Studies on the lysophosphatidylcholine metabolic enzymes revealed that phospholipase A (both phospholipase A1 and A2) activity was enhanced in the ethanol-perfused heart, but lysophospholipase and acyltransferase activities were unaffected by ethanol treatment. When the heart was perfused with 1% ethanol in the presence of 50-100 microM vitamin E, the ethanol-induced lysophosphatidylcholine accumulation was completely abolished. This was largely attributed to the attenuation of phospholipase A activities by vitamin E. In order to delineate the opposing effects of ethanol and vitamin E on phospholipid metabolism in the heart, phospholipase A activities in the subcellular fractions were determined in the presence of 0.5-2.0% ethanol or a combination of 1% ethanol and 0-100 microM vitamin E. Ethanol alone exhibited a biphasic effect on phospholipase A activity with maximum stimulation of enzyme activities at 1% concentration. When phospholipase A was assayed in 1% ethanol and vitamin E (25-100 microM), its activity was inhibited by vitamin E in a dose-dependent manner. The mechanism by which ethanol enhanced phospholipase A activities was further investigated with a partially purified enzyme from the rat heart cytosol. Kinetic studies with different concentrations of phosphatidylcholine revealed that at low substrate concentrations, ethanol was inhibitory to the reaction, whereas at high substrate concentrations, the reaction was enhanced by ethanol. Vitamin E (50 microM) completely abolished the ethanol-induced enhancement of enzyme activity in a noncompetitive manner. Since lysophosphatidylcholine is cytolytic at high concentration and its accumulation in the heart has been postulated as a biochemical cause of cardiac dysfunction, the level of the lysolipid in the heart must be under rigid control. Our result suggest that the modulation of cardiac phospholipase A activity is an important mechanism for the the regulation of lysophosphatidylcholine levels in the rat heart.
authors: Department of Biochemistry and Molecular Biology, University of Manitoba, Winnipeg, Canada.
authors: P C
authors: PC
authors: Choy
authors: K
authors: K
authors: O
authors: R Y
authors: RY
authors: Man
authors: A C
authors: AC
authors: Chan
mesh_terms: D000217:Acyltransferases / Q000378:metabolism; D000818:Animals; D000431:Ethanol / Q000494:pharmacology*; D006321:Heart / Q000187:drug effects*; D066298:In Vitro Techniques; D007700:Kinetics; D008245:Lysophospholipase / Q000378:metabolism; D008297:Male; D009097:Multienzyme Complexes / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D010477:Perfusion; D010713:Phosphatidylcholines / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism; D054466:Phospholipases A1; D010743:Phospholipids / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000378:metabolism; D014810:Vitamin E / Q000494:pharmacology*
chemical_list: D009097:Multienzyme Complexes; D010713:Phosphatidylcholines; D010743:Phospholipids; D014810:Vitamin E; D000431:Ethanol; D000217:Acyltransferases; D010741:Phospholipases A; D054466:Phospholipases A1; D008245:Lysophospholipase; C021782:lysophospholipase-transacylase