Hepatotoxicity of ethanol: protective effect of calcium channel blockers in isolated hepatocytes


Dpto. Pharmacology, University of Navarra, E-31080 Pamplona, Spain


Abstract: This study examines the effects of three calcium channel blockers (verapamil, nifedipine and diltiazem) on isolated rat hepatocytes exposed to ethanol. In the first part of our study, hepatocytes were incubated with increasing concentrations of ethanol (100, 300, 500, 1000 mM) for varying times. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) release were measured to evaluate the cytotoxic effects of ethanol. The concentration of 300 mM and time of incubation of 45 min were chosen for cytoprotection experiments in which calcium channel blockers, at two different concentrations, were added to the medium 30 min prior to the addition of ethanol. ALT, AST and LDH release as well as lipid peroxidation and cellular reduced gluthatione (GSH) were measured. Nifedipine and verapamil (25 μM) reduced ALT, AST and LDH activities. The highest dose of diltiazem (50 μM) was more effective than the lowest one (25 μM). Ethanol caused a significant depletion of cellular GSH content as well as a moderate enhancement of lipid peroxidation. While none of the three calcium channel blockers was able to restore the decrease in GSH levels, diltiazem (25 μM) and nifedipine (50 μM) showed the greatest effect, significantly reducing lipid peroxidation.