Secretion of cortisol and aldosterone as a vulnerable target for adrenal endocrine disruption — screening of 30 selected chemicals in the human H295R cell model

Abstract

The adrenal gland is a vulnerable target for toxic insult. Disruption of adrenal steroidogenesis and hormone secretion may cause serious effects on human health. A human in vitro model is needed to predict effects, and elucidate mechanisms of endocrine disruption and adrenal toxicity. The human adrenocortical cell line H295R has been used to screen for effects on sex hormones. Here, we have analyzed the effect of 30 potential endocrine disrupting chemicals on the secretion of cortisol and aldosterone from the H295R cells, using specific ELISA assays. The effect of chemicals was analyzed for basal and forskolin- or angiotensin II-stimulated hormone secretion. The chemicals were tested at the highest concentration where they displayed no evident unspecific cytotoxicity. Quantitative and qualitative differences in effects on hormone secretion were demonstrated for the various chemicals. A subset of the chemicals displayed different effects on cortisol and aldosterone secretion, and in some cases the effects were different between basal and stimulated hormone secretion. Aminoglutethimide, prochloraz, ketoconazole, 6-hydroxyflavone, imazalil and etomidate had the most marked inhibitory effects on cortisol (with or without forskolin) and ketoconazole, 6-hydroxyflavone, imazalil and etomidate had the most marked effects on aldosterone (with or without angiotensin II). The results are discussed in terms of known effects, structural similarity and possible mechanisms. We have shown that adrenal steroidogenesis is a vulnerable target for toxic insult and that the H295R assay is a useful in vitro model for screening purposes.

Mixture effects of imidazole fungicides on cortisol and aldosterone secretion in human adrenocortical H295R cells

“…This cell line is derived from a human adrenal tumor and has been used to investigate effects in vitro on hormone secretion following chemical treatment (Johansson et al, 2002;Oskarsson et al, 2006;Ullerås et al, 2008;Hecker and Giesy, 2008;Ohlsson et al, 2009). The cells express all steroidogenic enzymes and transport proteins required for synthesis and secretion of steroid hormones, including aldosterone and cortisol ( Fig.…”

Mitotane Effects in a H295R Xenograft Model of Adjuvant Treatment of Adrenocortical Cancer

“…The relative activities of the various steroidogenic CYP forms in H295R cells in vitro compared to the functional adrenal cortex in vivo are not exactly known, but could be diff erent. Eff ects on steroid production and CYP11 expression by drugs and xenobiotics has been examined [8,16,21,22] . The cell line is able to form grafts in immunodefi cient mice [23,24] .…”

“…The original culture was in suspension and subsequent selection produced the H295R variant, which grow in monolayer [15] . The cell line is known to express several steroidogenic enzymes [14 -18] , to secrete both aldosterone and cortisol [8,16] and to express a functional ACTH receptor [19,20] . The relative activities of the various steroidogenic CYP forms in H295R cells in vitro compared to the functional adrenal cortex in vivo are not exactly known, but could be diff erent.…”

“…The high adrenal toxicity of MeSO 2 -DDE is due to a local metabolic activation and irreversible metabolite binding in zona fasciculata, catalyzed by the zone-specifi c CYP11B1 enzyme [9 -12] . In vitro, MeSO 2 -DDE (7.5 μ M) has been shown to reduce cortisol secretion by 25 -50 % but was not observed to aff ect aldosterone secretion in monolayer cultured H295R cells [8] .…”

Effect of Polyphenols on Production of Steroid Hormones from Human Adrenocortical NCI-H295R Cells

“…1) Disrupting the secretion of these hormones has a serious impact on human health. For example, in Cushing's syndrome, hyperproduction of cortisol results in redistribution of fat to the face (moon face), hypertension, central obesity, and striae distensae.…”