Prenatal androgen excess alters the uterine peroxisome proliferator-activated receptor (PPAR) system.

It is known that androgen excess induces changes in fetal programming that affect several physiological pathways. Peroxisome proliferator-activated receptors (PPARs) α, δ and γ are key mediators of female reproductive functions, in particular in uterine tissues. Thus, we aimed to study the effect of prenatal hyperandrogenisation on the uterine PPAR system. Rats were treated with 2mg testosterone from Day 16 to 19 of pregnancy. Female offspring (PH group) were followed until 90 days of life, when they were killed. The PH group exhibited an anovulatory phenotype. We quantified uterine mRNA levels of PPARα (Ppara ), PPARδ (Ppard ), PPARγ (Pparg ), their regulators peroxisome proliferator-activated receptor gamma coactivator 1-alpha (Ppargc1a ) and nuclear receptor co-repressor 1 (Ncor1 ) and cyclo-oxygenase (COX)-2 (Ptgs2 ), and assessed the lipid peroxidation (LP) index and levels of glutathione (GSH) and prostaglandin (PG) E2 . The PH group showed decreased levels of all uterine PPAR isoforms compared with the control group. In addition, PGE2 and Ptgs2 levels were increased in the PH group, which led to a uterine proinflammatory environment, as was LP, which led to a pro-oxidant status that GSH was not able to compensate for. These results suggest that prenatal exposure to androgen excess has a fetal programming effect that affects the gene expression of PPAR isoforms, and creates a misbalanced oxidant-antioxidant state and a proinflammatory status.

Effect of hyperandrogenism on ovarian function.

The objective of this work was to study the ovarian function when follicular development is induced during a hyperandrogenic condition. Female rats were injected with either equine chorionic gonadotropin (eCG group) to induce folliculogenesis or eCG together with DHEA to induce folliculogenesis in a hyperandrogenic condition (eCG+HA group). The control group was injected with vehicle. Ovarian mRNA levels of the peroxisome proliferator-activated receptor gamma (PPARγ) co-activator PGC1α, the PPARγ co-repressor NCoR, the main enzymes involved in the ovarian steroidogenesis (CYP17, 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-HSD, and CYP19A), and cyclooxygenase 2 (COX2) were evaluated only by real-time PCR. COX2 was evaluated by both real-time PCR and western blot. Serum steroid hormones and both the oxidative and inflammatory statuses were also quantified. We found that eCG-induced folliculogenesis induced increased mRNA levels of PGC1α and decreased those of NCoR when compared with controls. In addition, we found an increase in serum estradiol (E2) levels and enhanced mRNA expression of CYP19A. A pro-inflammatory status and a pro-oxidant status were also established. When folliculogenesis was induced in a hyperandrogenic condition, the mRNA levels of the PPARγ co-repressor NCoR remained higher than in controls and the pro-inflammatory and pro-oxidant statuses were enhanced. In addition, the enzymes involved in ovarian steroidogenesis were altered leading to the accumulation of testosterone and an unfavorable E2/testosterone ratio. These alterations led to abnormal follicular development.

Prenatal hyperandrogenism and lipid profile during different age stages: an experimental study.

OBJECTIVE: The present study investigates the effect of prenatal hyperandrogenization on lipid metabolism and oxidant/antioxidant balance. DESIGN: Experimental study. SETTING: Research institute. ANIMAL(S): Pregnant Sprague Dawley rats were subcutaneously injected with 2 mg free T between days 16 and 19 of pregnancy, and controls (C) received vehicle (0.1 mL of sesame oil). Prenatally hyperandrogenized female offspring (T2) had a condition that resembles polycystic ovary (PCO). Animals were weighed and killed at 21 and 60 days of age (N = 15 rats/group). INTERVENTION(S): Ovarian tissue and truncal blood were obtained from the C and T2 groups. MAIN OUTCOME MEASURE(S): Circulating lipid profile (total cholesterol, high-density lipoprotein [HDL], low-density lipoprotein [LDL] cholesterol, and triglycerides) was quantified by colorimetric-enzymatic methods. Ovarian oxidative stress was evaluated by quantifying lipid peroxidation and glutathione content by spectofotometric assays. Ovarian fat content was evaluated by Red Oil staining and ovarian messenger RNA (mRNA) expression of peroxisome proliferator-activated receptor gamma (PPAR-γ) by real-time polymerase chain reaction (PCR). RESULT(S): At 60 days of age, 100% of group C rats and 20% of group T2 rats ovulated. At 21 days of age the T2 rats displayed lower body weight than C rats; however, at 60 days of age T2 and C rats showed similar body weights. The lipid profile (total cholesterol, LDL cholesterol, HDL cholesterol, and triglycerides) was altered in the anovulatory and ovulatory phenotype of the T2 group, but the levels were higher in the anovulatory phenotype. Lipid peroxidation of rats at 21 and 60 days of age from T2 was similar to C but the antioxidant glutathione level was decreased in 21-day-old rats compared with C rats. The lipid content of ovarian tissue, determined by Red Oil staining, was higher in the T2 than in the C group. The mRNA expression of ovarian PPAR-γ, quantified by real time PCR, decreased in anovulatory rats at 60 days of age from T2 compared to C rats. CONCLUSION(S): Our findings reveal the importance of evaluating the complete lipid profile, especially at early stages of life after the prenatal hyperandrogenism condition. In addition, we demonstrated that the antioxidant-reduced glutathione would represent a good marker of oxidative stress as it is altered before lipid peroxidation. Prenatal hyperandrogenization also alters the gene expression of PPAR-γ in rats. Here we demonstrated for the first time that abnormalities in PPAR-γ and lipid profile were higher in rats showing an anovulatory phenotype than those displaying an ovulatory phenotype.

[The role of prenatal hyperandrogenism on lipid metabolism during adult life in a rat model]. / Hiperandrogenización prenatal en ratas y el metabolismo lipídico en la vida adulta.

Polycystic ovary syndrome (PCOS) is one of the commonest endocrine diseases that affect women in their reproductive ages; however, the etiology of the syndrome remains unknown. A hypothesis proposes that during gestation increased exposure of androgen would induce fetal programming that may increase the risk of PCOS development during the adult life. By means of a prenatally hyperandrogenized (HA) rat model we demonstrated the importance of determining the lipid profile at early ages. HA induced two different phenotypes: ovulatory and anovulatory PCOS. HA did not modify total cholesterol but decreased HDL cholesterol and increased both LDL and tryglicerides (TG) when compared with controls. Both, the ratio total cholesterol: HDL (marker of cardiovascular risk) and TG:HDL (marker of metabolic syndrome) were increased in the HA group with respect to controls. In addition, these abnormalities were stronger in the anovulatory than ovulatory phenotype. Our results point out the need to find early markers of PCOS in girls or adolescents with increased risk to develop PCOS (as in daughters of women with PCOS).

Hiperandrogenización prenatal en ratas y el metabolismo lipídico en la vida adulta / The role of prenatal hyperandrogenism on lipid metabolism during adult life in a rat model

El síndrome del ovario poliquístico (PCOS) es una afección de alta incidencia en mujeres en edad fértil. Si bien la etiología de la enfermedad se desconoce, se cree que la exposición a andrógenos durante la vida intrauterina generaría reprogramación fetal afectando vías endocrinas y metabólicas que, junto a alteraciones génicas y ambientales, inducirían la aparición de PCOS en etapas muy tempranas de la vida. Es por ello que se buscan marcadores tempranos del desarrollo de PCOS. Utilizando un modelo murino de hiperandrogenización prenatal (HA) recreamos dos fenotipos de PCOS: ovulatorio y anovulatorio. La HA no alteró el colesterol circulante pero disminuyó el colesterol HDL y aumentó el LDL y los triglicéridos (TG) con respecto a los controles. La relación colesterol total/HDL como marcador de riesgo cardiovascular y la relación TG/HDL se vieron incrementadas con respecto a los controles, resultando mayor en el grupo PCOS anovulatorio. El presente trabajo demuestra la importancia de la determinación del perfil lipídico a edades tempranas en poblaciones de riesgo (como es el caso de hijas de madres con PCOS).

Polycystic ovary syndrome (PCOS) is one of the commonest endocrine diseases that affect women in their reproductive ages; however, the etiology of the syndrome remains unknown. A hypothesis proposes that during gestation increased exposure of androgen would induce fetal programming that may increase the risk of PCOS development during the adult life. By means of a prenatally hyperandrogenized (HA) rat model we demonstrated the importance of determining the lipid profile at early ages. HA induced two different phenotypes: ovulatory and anovulatory PCOS. HA did not modify total cholesterol but decreased HDL cholesterol and increased both LDL and tryglicerides (TG) when compared with controls. Both, the ratio total cholesterol: HDL (marker of cardiovascular risk) and TG:HDL (marker of metabolic syndrome) were increased in the HA group with respect to controls. In addition, these abnormalities were stronger in the anovulatory than ovulatory phenotype. Our results point out the need to find early markers of PCOS in girls or adolescents with increased risk to develop PCOS (as in daughters of women with PCOS).

Hiperandrogenización prenatal en ratas y el metabolismo lipídico en la vida adulta / The role of prenatal hyperandrogenism on lipid metabolism during adult life in a rat model

El síndrome del ovario poliquístico (PCOS) es una afección de alta incidencia en mujeres en edad fértil. Si bien la etiología de la enfermedad se desconoce, se cree que la exposición a andrógenos durante la vida intrauterina generaría reprogramación fetal afectando vías endocrinas y metabólicas que, junto a alteraciones génicas y ambientales, inducirían la aparición de PCOS en etapas muy tempranas de la vida. Es por ello que se buscan marcadores tempranos del desarrollo de PCOS. Utilizando un modelo murino de hiperandrogenización prenatal (HA) recreamos dos fenotipos de PCOS: ovulatorio y anovulatorio. La HA no alteró el colesterol circulante pero disminuyó el colesterol HDL y aumentó el LDL y los triglicéridos (TG) con respecto a los controles. La relación colesterol total/HDL como marcador de riesgo cardiovascular y la relación TG/HDL se vieron incrementadas con respecto a los controles, resultando mayor en el grupo PCOS anovulatorio. El presente trabajo demuestra la importancia de la determinación del perfil lipídico a edades tempranas en poblaciones de riesgo (como es el caso de hijas de madres con PCOS).(AU)

Polycystic ovary syndrome (PCOS) is one of the commonest endocrine diseases that affect women in their reproductive ages; however, the etiology of the syndrome remains unknown. A hypothesis proposes that during gestation increased exposure of androgen would induce fetal programming that may increase the risk of PCOS development during the adult life. By means of a prenatally hyperandrogenized (HA) rat model we demonstrated the importance of determining the lipid profile at early ages. HA induced two different phenotypes: ovulatory and anovulatory PCOS. HA did not modify total cholesterol but decreased HDL cholesterol and increased both LDL and tryglicerides (TG) when compared with controls. Both, the ratio total cholesterol: HDL (marker of cardiovascular risk) and TG:HDL (marker of metabolic syndrome) were increased in the HA group with respect to controls. In addition, these abnormalities were stronger in the anovulatory than ovulatory phenotype. Our results point out the need to find early markers of PCOS in girls or adolescents with increased risk to develop PCOS (as in daughters of women with PCOS).(AU)