Mono-(2-ethylhexyl) phthalate stimulates basal steroidogenesis by a cAMP-independent mechanism in mouse gonadal cells of both sexes.
Phthalates are widely used as plasticizers in a number of daily-life products. In this study, we investigated the influence of mono-(2-ethylhexyl) phthalate (MEHP), the active metabolite of the frequently used plasticizer di-(2-ethylhexyl) phthalate (DEHP), on gonadal steroidogenesis in vitro. MEHP (25-100 microM) stimulated basal steroid synthesis in a concentration-dependent manner in immortalized mouse Leydig tumor cells (MLTC-1). The stimulatory effect was also detected in KK-1 granulosa tumor cells. MEHP exposure did not influence cAMP or StAR protein levels and induced a gene expression profile of key steroidogenic proteins different from the one induced by human chorionic gonadotropin (hCG). Simultaneous treatment with MEHP and a p450scc inhibitor (aminoglutethimide) indicated that MEHP exerts its main stimulatory effect prior to pregnenolone formation. MEHP (10-100 microM) up-regulated hormone-sensitive lipase and 3-hydroxy-3-methylglutaryl coenzyme A reductase, suggesting that MEHP increases the amount of cholesterol available for steroidogenesis. Our data suggest that MEHP, besides its known inhibitory effect on hCG action, can directly stimulate gonadal steroidogenesis in both sexes through a cAMP- and StAR-independent mechanism. The anti-steroidogenic effect of DEHP has been proposed to cause developmental disorders such as hypospadias and cryptorchidism, whereas a stimulation of steroid synthesis may prematurely initiate the onset of puberty and theoretically affect the hypothalamic-pituitary-gonadal axis.
Gunnarsson
David
D
Department of Molecular Biology, Umeå University, S-901 87 Umeå, Sweden. gunnar.selstam@molbiol.umu.se
Leffler
Per
P
Ekwurtzel
Emelie
E
Martinsson
Gunilla
G
Liu
Kui
K
Selstam
Gunnar
G
D000616:Aminoglutethimide / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D004051:Diethylhexyl Phthalate / Q000009:adverse effects*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006066:Gonads; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism; D007985:Leydig Cells / Q000187:drug effects / Q000378:metabolism*; D008049:Lipase / Q000378:metabolism; D008297:Male; D051379:Mice; D010750:Phosphoproteins / Q000378:metabolism; D010968:Plasticizers / Q000009:adverse effects*; D011374:Progesterone / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013268:Stimulation, Chemical; D013739:Testosterone / Q000096:biosynthesis*; D000098682:Steroidogenic Acute Regulatory Protein
D004791:Enzyme Inhibitors; D010750:Phosphoproteins; D010968:Plasticizers; D000098682:Steroidogenic Acute Regulatory Protein; D000616:Aminoglutethimide; D013739:Testosterone; D011374:Progesterone; D004051:Diethylhexyl Phthalate; D006903:Hydroxymethylglutaryl CoA Reductases; D008049:Lipase