Amphetamine-decreased progesterone and estradiol release in rat granulosa cells: The regulatory role of camp-and ca²⁺-mediated signaling pathways

The purpose of this study is to evaluate the amphetamine effects on progesterone and estradiol production in rat granulosa cells and the underlying cellular regulatory mechanisms. Freshly dispersed rat granulosa cells were cultured with various test drugs in the presence of am-phetamine, and the estradiol/progesterone production and the cytosolic cAMP level were meas-ured. Additionally, the cytosolic-free Ca²⁺ concentrations ([Ca²⁺]i) were measured to examine the role of Ca²⁺ influx in the presence of amphetamine. Amphetamine in vitro inhibited both basal and porcine follicle-stimulating hormone-stimulated estradiol/progesterone release, and amphetamine significantly decreased steroidogenic enzyme activities. Adding 8-Bromo-cAMP did not recover the inhibitory effects of amphetamine on progesterone and estradiol release. H89 significantly decreased progesterone and estradiol basal release but failed to enhance a further amphetamine in-hibitory effect. Amphetamine was capable of further suppressing the release of estradiol release under the presence of nifedipine. Pretreatment with the amphetamine for 2 h decreased the basal [Ca²⁺]i and prostaglandin F2α-stimulated increase of [Ca²⁺]i. Amphetamine inhibits progesterone and estradiol secretion in rat granulosa cells through a mechanism involving decreased PKA-down-stream steroidogenic enzyme activity and L-type Ca²⁺ channels. Our current findings show that it is necessary to study the possibility of amphetamine perturbing reproduction in females.