The effect of valproate and levetiracetam on steroidogenesis in forskolin-stimulated H295R cells.
OBJECTIVE
Endocrine disruptive effects have been frequently observed in patients using antiepileptic drugs (AEDs). Two different AEDs, valproate (VPA) and levetiracetam (LEV), were tested in forskolin-stimulated human adrenal carcinoma (H295R) cells to explore their effect on steroidogenesis. VPA has a long history as an anticonvulsant and is linked with many of the endocrine disorders associated with AED use. LEV is a newer AED, and no endocrine disruptive effects have been reported in humans to date.


METHODS
H295R cells, which are capable of full steroidogenesis, were stimulated with forskolin and exposed to either VPA or LEV for 48 h. Medium was collected and analyzed for hormone production. For the VPA-exposed cells, steroidogenic gene expression analysis was also conducted.


RESULTS
VPA exposure resulted in a significant reduction in progesterone and estradiol (E2) production, whereas testosterone (T) levels remained unchanged. There were also significant alterations in expression level for most genes analyzed. LEV exposure resulted in a minor, but statistically significant, reduction in T and E2 production.


CONCLUSIONS
Exposure of forskolin-stimulated H295R cells to VPA led to an increased T/E2 ratio through a significant decrease in estradiol production. Gene analysis suggested that VPA affects NR0B1 expression. NR0B1 inhibits promoters of other genes involved in steroidogenesis, and the altered expression of NR0B1 might explain the observed down-regulation in hormone production. The effects of LEV exposure on hormone secretion were not considered to be biologically significant.
von Krogh
Kristine
K
Department of Production Animal Clinical Sciences, Norwegian School of Veterinary Science, Oslo, Norway.
Harjen
Hannah
H
Almås
Camilla
C
Zimmer
Karin E
KE
Dahl
Ellen
E
Olsaker
Ingrid
I
Taubøll
Erik
E
Ropstad
Erik
E
Verhaegen
Steven
S
D000310:Adrenal Gland Neoplasms / Q000473:pathology; D000927:Anticonvulsants / Q000494:pharmacology*; D001141:Aromatase / Q000378:metabolism; D045744:Cell Line, Tumor; D005576:Colforsin / Q000494:pharmacology*; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics; D057137:DAX-1 Orphan Nuclear Receptor / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D000077287:Levetiracetam; D010889:Piracetam / Q000031:analogs & derivatives* / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000378:metabolism*; D054339:Steroidogenic Factor 1 / Q000235:genetics; D013268:Stimulation, Chemical; D013739:Testosterone / Q000378:metabolism*; D014635:Valproic Acid / Q000494:pharmacology*
D000927:Anticonvulsants; D057137:DAX-1 Orphan Nuclear Receptor; C539965:NR0B1 protein, human; C515057:NR5A1 protein, human; D054339:Steroidogenic Factor 1; D005576:Colforsin; D013739:Testosterone; D000077287:Levetiracetam; D011374:Progesterone; D004958:Estradiol; D014635:Valproic Acid; D001141:Aromatase; D019363:Cytochrome P-450 CYP1A1; D010889:Piracetam