Abstract
Anabolic–androgenic steroids are testosterone derivatives, used by body-builders to increase muscle mass. Epistane (EPI) is an orally administered 17α-alkylated testosterone derivative with 2a-3a epithio ring. We identified four individuals who, after EPI consumption, developed long-lasting cholestasis. The bile acid (BA) profile of three patients was characterized, as well the molecular mechanisms involved in this pathology. The serum BA pool was increased from 14 to 61-fold, basically on account of primary conjugated BA (cholic acid (CA) conjugates), whereas secondary BA were very low. In in vitro experiments with cultured human hepatocytes, EPI caused the accumulation of glycoCA in the medium. Moreover, as low as 0.01 μM EPI upregulated the expression of key BA synthesis genes (CYP7A1, by 65% and CYP8B1, by 67%) and BA transporters (NTCP, OSTA and BSEP), and downregulated FGF19. EPI increased the uptake/accumulation of a fluorescent BA analogue in hepatocytes by 50–70%. Results also evidenced, that 40 μM EPI trans-activated the nuclear receptors LXR and PXR. More importantly, 0.01 μM EPI activated AR in hepatocytes, leading to an increase in the expression of CYP8B1. In samples from a human liver bank, we proved that the expression of AR was positively correlated with that of CYP8B1 in men. Taken together, we conclude that EPI could cause cholestasis by inducing BA synthesis and favouring BA accumulation in hepatocytes, at least in part by AR activation. We anticipate that the large phenotypic variability of BA synthesis enzymes and transport genes in man provide a putative explanation for the idiosyncratic nature of EPI-induced cholestasis.
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Acknowledgements
This research was supported by an EU grant (7th Framework Program; project acronym: HeCaToS “Hepatic and Cardiac Toxicity Systems modelling” Grant Agreement No: 602156) which allowed the creation of a Clinical Hepatotoxicity Unit at the Hospital La Fe led by IC, and by Instituto de Salud Carlos III through the project “PI17/01089” (co-founded by European Regional Development Fund “A way to achieve Europe”). PDP was the holder of a research contract sponsored by CIBEREHD (National Biomedical Research Institute on Liver and Gastrointestinal Diseases, Instituto de Salud Carlos III, Spain) and is currently supported by Sara Borrell Postdoctoral Contract CD18/00158 from the Instituto de Salud Carlos III, co-founded by the European Social Fund.
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Petrov, P.D., Fernández-Murga, L., Conde, I. et al. Epistane, an anabolic steroid used for recreational purposes, causes cholestasis with elevated levels of cholic acid conjugates, by upregulating bile acid synthesis (CYP8B1) and cross-talking with nuclear receptors in human hepatocytes. Arch Toxicol 94, 589–607 (2020). https://doi.org/10.1007/s00204-019-02643-y
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DOI: https://doi.org/10.1007/s00204-019-02643-y