Effects of trichlorfon on progesterone production in cultured human granulosa-lutein cells

Abstract

Trichlorfon, a widely used organophophorus pesticide, has been reported to disrupt reproductive function in human and animal. However, the mechanisms have not been clearly elucidated. In the present study, the effects of trichlorfon on the biosynthesis of progesterone in the primary human granulosa-lutein cells (hGLCs) and the related pathway were investigated. Results showed that progesterone production in hGLCs treated with trichlorfon decreased significantly while cell viability was not affected. Trichlorfon also inhibited FSH-stimulated progesterone production, and this inhibition could not be reversed by 8-bromo-cAMP. However, trichlorfon did not affect the intracellular cAMP contents in the basal and FSH-stimulated conditions. These results suggested that the site in the steroid biosynthesis pathway affected by trichlorfon occurred downstream of PKA activation in hGLCs. Furthermore, our results found that 22(R)-hydroxycholesterol (22R-HC) could remove the inhibitory action of trichlorfon on progesterone biosynthesis, indicating that trichlorfon caused a disruption of cholesterol transport across mitochondrial membranes, which was further confirmed by the observation that trichlorfon dose-dependently inhibited the mRNA level of the steroidogenic acute regulatory protein (StAR). These results suggested that trichlorfon inhibited steroidogenesis in hGLCs by reducing StAR gene expression, which may further contribute to the pathogenesis of trichlorfon-induced reproductive dysfunction.