EFFECT OF ALCOHOL ON EXPRESSION OF HEAT-SHOCK PROTEIN-73 IN MONOLAYERS OF HEPATOCYTES FROM ALCOHOL-EXPOSED RATS

Alcohol dramatically reduced loss of heat shock proteins (HSP73) and p revented morphological damage in monolayers of hepatocytes prepared fr om alcohol-fed rats. The monolayers were treated with 0, 5, 25, or 100 mM alcohol and triplicate samples were assayed at 24, 48, 72, and 96 hr after exposure. The content of HSP73 was measured by PAGE electroph oresis and Western blotting with a mouse monoclonal anti-HSP70 IgG ant ibody. HSP72 is not expressed under these conditions. Damage to the he patocytes, quantified by leakage of lactate dehydrogenase (LDH), was a lso decreased by 100 mM alcohol. Although the initial 100 mM alcohol c oncentration decreased logarithmically to 1.7 mM over the first 24 hr, the effect of alcohol on HSP73 loss, LDH leakage, and morphological d amage was most pronounced at 96 hr. (C) 1994 Academic Press, Inc.