@article {Sharanekdmd.117.077420, author = {Ahmad Sharanek and Audrey Burban and Lydie Humbert and Christiane Guguen-Guillouzo and Dominique Rainteau and Andre Guillouzo}, title = {Progressive and preferential cellular accumulation of hydrophobic bile acids induced by cholestatic drugs is associated with inhibition of their amidation and sulfation}, elocation-id = {dmd.117.077420}, year = {2017}, doi = {10.1124/dmd.117.077420}, publisher = {American Society for Pharmacology and Experimental Therapeutics}, abstract = {Drug-induced intrahepatic cholestasis is characterized by cellular accumulation of bile acids (BA) whose mechanisms remain poorly understood. The present study aimed to analyze early and progressive alterations of BA profiles induced by cyclosporine A, chlorpromazine, troglitazone, tolcapone, trovafloxacin and tacrolimus after 4h, 24h and 6 daily treatments of differentiated HepaRG cells. In BA-free medium the potent cholestatic drugs, cyclosporine A, chlorpromazine and troglitazone, reduced endogenous BA synthesis after 24h, while the rarely cholestatic drugs, tolcapone, trovafloxacin and tacrolimus, reduced BA synthesis only after 6 days. In the presence of physiological serum BA concentrations, cyclosporine A, chlorpromazine and troglitazone induced early and preferential cellular accumulation of unconjugated lithocholic, deoxycholic and chenodeoxycholic acids that increased 8-12- and 47-50-fold after 24h and 6 days respectively. Accumulation of these hydrophobic BAs resulted from strong inhibition of amidation and in addition, for lithocholic acid reduction of its sulfo-conjugation, and was associated with variable alterations of uptake and efflux transporters. Trovafloxacin also caused BA accumulation, especially after 6 days, while tolcapone and tacrolimus were still without effect. However, when exogenous BAs were added to the medium at cholestatic serum concentrations, a 6-day treatment with all drugs resulted in cellular BA accumulation with higher folds of chenodeoxycholic and lithocholic acids. At the tested concentration, tolcapone had the lowest effect. These results bring the first demonstration that major cholestatic drugs can cause preferential and progressive in vitro cellular accumulation of unconjugated toxic hydrophobic BAs, and bring new insights in mechanisms involved in drug-induced cellular accumulation of toxic BAs.}, issn = {0090-9556}, URL = {https://dmd.aspetjournals.org/content/early/2017/09/19/dmd.117.077420}, eprint = {https://dmd.aspetjournals.org/content/early/2017/09/19/dmd.117.077420.full.pdf}, journal = {Drug Metabolism and Disposition} }