Corticosteroid Production in H295R Cells During Exposure to 3 Endocrine Disrupters Analyzed With LC-MS/MS

International Journal of Toxicology - Tập 32 Số 3 - Trang 219-227 - 2013
Christina S. Winther1, Frederik Nielsen1, Martin Hansen1, Bjarne Styrishave1
1Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken, Copenhagen, Denmark

Tóm tắt

The adrenocortical human cell line H295R is a valuable tool for screening endocrine disrupting compounds. In general, previous research focus has been on the production of the 2 sex steroids, 17β-estradiol and testosterone, and less attention has been paid to other important steroid end points in the steroidogenesis with a wide range of physiological functions, such as the glucocorticoids (corticosterone and cortisol). A newly developed and validated solid phase extraction (SPE) liquid chromatography–mass spectroscopy (LC-MS/MS) method was used to measure the production of cortisol and corticosterone in the H295R cell line. The method was applied by studying the effects of 2 model endocrine disrupters, ketoconazole and prochloraz, the pharmaceutical budesonide, and the inducer forskolin on the steroid production in this cell line. Dose–response curves were obtained for the correlation between hormone concentrations and the concentration of the individual disruptors. Exposing cells to ketoconazole resulted in a decrease in cortisol and corticosterone concentrations in a dose-dependent manner with EC50 values of 0.24 and 0.40 μmol/L, respectively. The same applied for cells exposed to prochloraz with EC50 values of 0.06 and 0.09 μmol/L for cortisol and corticosterone, respectively. Budesonide also inhibited glucocorticoid secretion. The EC50 value for cortisol was 19.50 μmol/L, whereas the EC50 value for corticosterone was 71.42 μmol/L. Forskolin induced the secretion of both cortisol (EC50 = 4.09 μmol/L) and corticosterone (EC50 = 0.28 μmol/L). The results obtained demonstrated the validity of the method. Based on these findings, quality criteria for the production of these steroids in this cell line were suggested.

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International Journal of Toxicology

Tập 32 Số 3

219-227

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