Study on ketoconazole-induced hepatotoxicity and its mechanisms in a primary cultured rat hepatocytes system
Liao Ming-yang · Journal of Toxicology · 2006
Objective To observe the effects of ketoconazote on cell livability,lactate dehydrogenawe(LDH) leakage and the contents of sulfhydryl groups in a primary cultured hepatocytes system of Sprague-Dawley rat and to investigate the potential hepatoxic mechanisms of ketoconazole.Methods Hepatocytes were isolated from male adult Sprague-Dawley rats by the two-step perfusion using collagenase and cultured in William's E culture medium with 10% fetal serum.Rat hepatocytes were administered with different doses(0,56,75,94,113 and 188 μmnol/L) of ketoconazole for 4 h or with 188 μmol/L for different time(0,1,2 and 4 h) to investigate dose-effect and time-effect relationships by measuring the leakage of the LDH into the medium;by assessing mitochondrial reduction of 3-(4,5-dimethythiazol-2yl)-2,5-diphenyl tetrazolium bromide(MTT) and lysosomal uptake of neutral red(NR);and by ssessing the content of total sulfhydryl and non-protein binding sulfhydryl.Results 1.cell livability and the content of sulfhydryl groups declined and LDH leakage increased significantly with the increase of the administered dose.2.cell livability and the content of sulfhydryl groups declined and LDH leakage increased significantly with the prolongation of the administered dose.Conclusion These results demonstrate that the hepatoxicity of ketoconazole is related with the alterations of the contents of sulfhydryl groupsintra-cellular.