The following is a summary of “Extended steroid profiling in H295R cells provides deeper insight into chemical-induced disturbances of steroidogenesis: Exemplified by prochloraz and anabolic steroids,” published in the June 2023 issue of Molecular and Cellular Endocrinology by Jäger et al.

OECD Test Guideline 456 has validated human adrenocortical H295R cells for detecting compounds that inhibit testosterone and 17-estradiol (estradiol) biosynthesis. This study evaluated a novel method for detecting steroidogenesis disturbances in H295R cells, as illustrated by prochloraz and five anabolic steroids. An untargeted LC-MS method assessed steroid profiles, yielding relative quantifications of 57 steroids annotated with their accurate masses and retention times. 

Several mineralocorticoids, glucocorticoids, progestins, and adrenal androgens comprised this panel of steroids. This extensive steroid profiling enabled the clustering of chemicals with similar effects and the detection of subtle differences between chemicals by encompassing many metabolites. It made it possible, for instance, to distinguish between the impact of turinabol and oxymetholone, which were assumed to act similarly in a previous classification that included only nine adrenal steroids. In addition, the outcomes demonstrated that product/substrate ratios provide superior information on altered enzyme activities compared to individual metabolite levels. 

For instance, it was discovered that the 17-hydroxypregnenolone/pregnenolone ratio is a more sensitive marker for detecting 17-hydroxylase inhibition by prochloraz than the corresponding individual steroids. These results demonstrate that chemical classification and the calculation of product/substrate ratios can provide helpful information on the mode of action and aid in prioritizing future experimental work.

Source: sciencedirect.com/science/article/pii/S0303720723000801