Bovine liver slices combined with an androgen transcriptional activation assay: an in-vitro model to study the metabolism and bioactivity of steroids

Wang, S.; Rijk, J.; Riethoff-Poortman, J.; Kuijk, S.; Peijnenburg, A.; Bovee, T.
May 2010
Analytical & Bioanalytical Chemistry;May2010, Vol. 397 Issue 2, p631
Academic Journal
Previously we described the properties of a rapid and robust yeast androgen bioassay for detection of androgenic anabolic compounds, validated it, and showed its added value for several practical applications. However, biotransformation of potent steroids into inactive metabolites, or vice versa, is not included in this screening assay. Within this context, animal-friendly in-vitro cellular systems resembling species-specific metabolism can be of value. We therefore investigated the metabolic capacity of precision-cut slices of bovine liver using 17�-testosterone (T) as a model compound, because this is an established standard compound for assessing the metabolic capacity of such cellular systems. However, this is the first time that slice metabolism has been combined with bioactivity measurements. Moreover, this study also involves bioactivation of inactive prohormones, for example dehydroepiandrosterone (DHEA) and esters of T, and although medium extracts are normally analyzed by HPLC, here the metabolites formed were identified with more certainty by ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC�TOFMS) with accurate mass measurement. Metabolism of T resulted mainly in the formation of the less potent phase I metabolites 4-androstene-3,17-dione (4-AD), the hydroxy-T metabolites 6a, 6�, 15�, and 16a-OH-T, and the phase II metabolite T-glucuronide. As a consequence the overall androgenic activity, as determined by the yeast androgen bioassay, decreased. In order to address the usefulness of bovine liver slices for activation of inactive steroids, liver slices were exposed to DHEA and two esters of T. This resulted in an increase of androgenic activity, because of the formation of 4-AD and T. [Figure not available: see fulltext.]


Related Articles

  • Evidence of the indirect hormonal activity of prohormones using liver S9 metabolic bioactivation and an androgen bioassay. Rijk, J. C. W.; Bovee, T. F. H.; Groot, M. J.; Peijnenburg, A. A. C. M.; Nielen, M. W. F. // Analytical & Bioanalytical Chemistry;Oct2008, Vol. 392 Issue 3, p417 

    Prohormones such as dehydroepiandrosterone (DHEA) are steroid precursors that do not show hormonal activity by themselves. Abuse of these prohormones in cattle fattening is hard to prove because of strong in vivo metabolism and the difficulty to detect metabolites which are not significantly...

  • Preclinical pharmacological profile of nomegestrol acetate, a synthetic 19-nor-progesterone derivative. van Diepen, Harry A. // Reproductive Biology & Endocrinology;2012, Vol. 10 Issue 1, p1 

    Background: Nomegestrol acetate (NOMAC), a synthetic progestogen derived from 19-nor-progesterone, recently completed clinical trials for use with 17beta-estradiol in a new monophasic combined oral contraceptive. In this review, published as well as previously unpublished preclinical studies...

  • Implication of human UGT2B7, 2B15, and 2B17 in 19-norandrosterone metabolism. Strahm, Emmanuel; Sjöberg, Ulf; Garle, Mats; Rane, Anders; Ekström, Lena // Frontiers in Endocrinology;Jun2013, Vol. 4, p1 

    Nandrolone (19-nortestosterone) is an anabolic androgenic steroid commonly abused for doping purposes. Nandrolone is mainly metabolized in the liver into 19-norandrosterone prior to glucuronidation and excretion through urine over an extended period of time. Several UGTs (i.e., UGT2B7, UGT2B15,...

  • Steroidogenic enzyme expression in the human fetal liver and potential role in the endocrinology of pregnancy. O'Shaughnessy, P.J.; Monteiro, A.; Bhattacharya, S.; Fraser, M.J.; Fowler, P.A. // MHR: Basic Science of Reproductive Medicine;Mar2013, Vol. 19 Issue 3, p177 

    The human feto-maternal unit produces large amounts of steroid hormones, particularly estrogens, during the second and third trimesters. The fetal adrenal gland and the placenta are considered the principal tissues driving steroid production but the fetal liver is likely to play an essential...

  • The impact of PPARα activation on whole genome gene expression in human precision cut liver slices. Janssen, Aafke W. F.; Betzel, Bark; Stoopen, Geert; Berends, Frits J.; Janssen, Ignace M.; Peijnenburg, Ad A.; Kersten, Sander // BMC Genomics;10/9/2015, Vol. 16 Issue 1, p1 

    Background: Studies in mice have shown that PPARα is an important regulator of lipid metabolism in liver and key transcription factor involved in the adaptive response to fasting. However, much less is known about the role of PPARα in human liver. Methods: Here we set out to study the...

  • Liver slices are the optimal model for mimicking apoptosis activation in vitro. Milisav, Irina; Supul, Dusan // HealthMed;2011 Supplement 1, Vol. 5, p2047 

    Tissue slicing enables sectioning of tissue with metabolically active cells. The cutting of untreated rat and human liver tissues has been optimized recently to produce uniformly thick tissue slices of 200 p.m. There are no reports in the literature about the optimal slice thickness to study...

  • Acquisition of Androgen Independence by Human Prostate Epithelial Cells during Arsenic-Induced Malignant Transformation. Benbrahim-Tallaa, Lamia; Webber, Mukta M.; Waalkes, Michael P. // Environmental Health Perspectives;Sep2005, Vol. 113 Issue 9, p1134 

    Lethal phenotypes of human prostate cancer are characterized by progression to androgen independence, although the mechanisms behind this progression remain unclear. Arsenic is a potential human prostate carcinogen that may affect tumor progression. In this study, we used a prostate cancer...

  • Enhancement of steroid receptor-mediated transcription for the development of highly responsive bioassays. Willemsen, Philippe; Scippo, Marie-Louise; Maghuin-Rogister, Guy; Martial, Joseph A.; Muller, Marc // Analytical & Bioanalytical Chemistry;Jun2005, Vol. 382 Issue 4, p894 

    We have previously generated several transformed human mammary cell lines for the detection of steroid receptor-mediated activities and used these cell lines to detect and characterize steroid hormone (ant)agonistic compounds. In this report, we describe the specific optimization procedures used...

  • Altered Balance Between the 5alpha-Reductase and Aromatase Pathways of Androgen Metabolism.... Slater, Cristin C.; Chang, Lilly; Stanczyk, Frank Z.; Paulson, Richard J. // Journal of Assisted Reproduction & Genetics;Oct2001, Vol. 18 Issue 10, p527 

    Examines androgen production and metabolism during controlled ovarian hyperstimulation. Increase in steroid level from baseline during controlled ovarian hyperstimulation; Decrease in 5alpha-reductase activity; Enhancement in androgen metabolism by aromatase.


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics