Putative activation of the peroxisome proliferator-activated receptor gamma impairs androgen and enhances progesterone biosynthesis in primary cultures of porcine theca cells.
Ovarian theca cells are the predominant source of gonadotropin-stimulated androgen biosynthesis in vivo. Troglitazone (TG), a synthetic agonist of the peroxisome proliferator-activated receptor gamma (PPARgamma) and a thiazolidinedione used to treat insulin resistance, decreases serum androgen concentrations in women with hyperthecosis and/or polycystic ovary syndrome. Using reverse transcription-polymerase chain reaction (RT-PCR), we demonstrated the presence of PPARgamma mRNA in the porcine ovary. Since activation of ovarian PPARgamma may alter hormone-stimulated steroidogenesis in vitro, we cultured porcine theca cells for 48 h in the presence of two different PPARgamma ligands, TG and 15-deoxy-Delta(12,14)-prostaglandin J2 (15d-PGJ2). Putative TG-mediated activation of PPARgamma resulted in a 53%-69% decrease in LH- and/or insulin-stimulated androstenedione and testosterone accumulation. Although TG reduced 3-isobutylmethylxanthine-enhanced LH-stimulated cAMP accumulation by 74%-78%, it did not alter basal cAMP concentrations. Exposure to 8Br-cAMP did not overcome the TG-induced inhibition of androgen accumulation. In contrast, TG administration amplified basal and hormone-stimulated progesterone accumulation, particularly in the presence of insulin, without altering levels of 17alpha-hydroxyprogesterone. The putative natural PPARgamma ligand, 15d-PGJ2, inhibited androgen biosynthesis and stimulated progesterone production. RT-PCR-based amplification of cytochrome P450 cholesterol side-chain cleavage (CYP11A) and cytochrome P450 17alpha-hydroxylase/C-17,20-lyase (CYP17) transcripts indicated that TG moderately enhanced expression of these genes. However, TG did not affect CYP17 protein expression. We conclude that putative ligand-mediated activation of PPARgamma decreases LH- and/or insulin-driven theca cell androgen production by impairing the ability of CYP17 to synthesize androstenedione from available progestins. The corresponding augmentation of progesterone production could suggest that PPARgamma activation induces theca cell differentiation toward a progestin-synthesizing phenotype.
Schoppee
Pamela D
PD
Division of Endocrinology and Metabolism, Department of Internal Medicine, University of Virginia Medical School, Charlottesville, Virginia 22908, USA.
Garmey
James C
JC
Veldhuis
Johannes D
JD
D000728:Androgens / Q000096:biosynthesis*; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D005456:Fluorescent Dyes; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000494:pharmacology; D009930:Organic Chemicals; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D011374:Progesterone / Q000096:biosynthesis*; D015230:Prostaglandin D2 / Q000031:analogs & derivatives* / Q000378:metabolism; D011863:Radioimmunoassay; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013254:Steroid 17-alpha-Hydroxylase / Q000096:biosynthesis / Q000235:genetics; D013552:Swine; D013799:Theca Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D013844:Thiazoles / Q000494:pharmacology; D045162:Thiazolidinediones; D014157:Transcription Factors / Q000378:metabolism*; D000077288:Troglitazone
C097240:15-deoxy-delta(12,14)-prostaglandin J2; D000728:Androgens; D002839:Chromans; D005456:Fluorescent Dyes; D007004:Hypoglycemic Agents; D007328:Insulin; C433144:NanoOrange; D009930:Organic Chemicals; C099476:PicoGreen; D018160:Receptors, Cytoplasmic and Nuclear; D013844:Thiazoles; D045162:Thiazolidinediones; D014157:Transcription Factors; D011374:Progesterone; D007986:Luteinizing Hormone; D004247:DNA; D013254:Steroid 17-alpha-Hydroxylase; D000077288:Troglitazone; D015230:Prostaglandin D2