Abstracts

Rationale: Endocrine disruptive effects have been frequently observed in patients using anti-epileptic drugs (AED). Two different AEDs were tested in forskolin-stimulated human adrenocarcinoma (H295R) cells to explore their effect on steroidogenesis. The AEDs tested were valproate (VPA), which has a long history as an anticonvulsant and is linked with many of the endocrine disorders associated with AED use, and levetiracetam (LEV), a newer AED, with no endocrine disruptive effects in humans described this far. Methods: H295R cells, which are capable of full steroidogenesis, were stimulated with forskolin and exposed to either VPA or LEV for 48 hours. Medium was collected and analyzed for hormone production (oestradiol, testosterone and progesterone). For the VPA exposed cells steroidogenic gene expression analysis was also conducted, by mRNA extraction and real time qPCR. Results: The cells showed increased testosterone production, but decreased oestradiol and progesterone production when exposed to VPA. Alteration in hormone production after VPA exposure occurred in a dose-dependent manner (p<0.05). After LEV exposure, the cells showed a small dose-dependent decrease in testosterone and oestradiol production (p<0.01), while progesterone production remained unchanged. Gene-expression analysis after VPA exposure showed a reduction in mRNA levels for almost all genes analyzed. Notably, DAX-1 expression levels were up-regulated, which might explain the observed down-regulation of other genes, such as CYP17, StAR and 3βHSD2. Conclusions: In forskolin-stimulated H295R cells, VPA and LEV exposure led to significant alterations in hormone production. Gene analysis after VPA exposure suggests that VPA affects DAX-1 expression. DAX-1 inhibits promoters of others genes involved in steroidogenesis, so this alteration in its expression might be the cause of down-regulation in hormone production observed in this study. The decreased testosterone and oestradiol production observed after LEV exposure was very small, but might suggest that LEV affects the expression of CYP17, 17βHSD1 or 3βHSD2.