Polyphenols and IUGR Pregnancies: Effects of Maternal Hydroxytyrosol Supplementation on Placental Gene Expression and Fetal Antioxidant Status, DNA-Methylation and Phenotype.

The use of polyphenols is a promising strategy for preventing or alleviating intrauterine growth restriction (IUGR) because polyphenol supplementation increases plasma antioxidant capacity and improves oxidative stress at the feto-placental unit; which are recognized as main issues in IUGR. However, there is a scarcity of experimental data on both realistic benefits and potential hazards of polyphenol supplementation during gestation. Hence, we aimed to use a swine model of IUGR pregnancy to determine possible effects of maternal supplementation with polyphenols (hydroxytyrosol) on placental expression of genes involved in antioxidant homeostasis, vascularization and fetal growth and thus on antioxidant status, DNA-methylation and phenotypic traits (morphology and homeostasis) of the fetus. Hydroxytyrosol improves placental gene expression and fetal antioxidant status and glucose metabolism in a sex-dependent manner, in which males were favored in spite of developmental failures. Concomitantly, hydroxytyrosol prevented hypomethylation of DNA associated with oxidative stress. Finally, no major deleterious effects of hydroxytyrosol supplementation on constriction of the ductus arteriosus, a possible secondary effect of polyphenols during pregnancy, were found.

Antioxidant homeostasis is disturbed in fetuses with leptin-resistant genotypes: A cross-sectional study.

BACKGROUND: Leptin resistance is associated with lower reproductive efficiency, with deficiencies in embryo viability and growth leading to low prolificacy and high incidence of intrauterine growth restriction. OBJECTIVE: We aimed to investigate the underlying mechanisms of the leptin-resistance, evaluating the antioxidant homeostasis of leptin-resistant and lean swine fetuses. MATERIALS AND METHODS: The study included 70 plasma samples from fetuses at day 62 of gestation (mid-pregnancy), from breeds with (Iberian breed; n=35) and without leptin resistance (25% Large White x 25% Landrace x 50% Pietrain; n=35). The antioxidant status of the plasma samples was determined by photoinduced chemiluminescence whilst systemic oxidative stress was assessed determining plasma hydrogen peroxide concentration by enzimoimmunoassay. RESULTS: Plasma total antioxidant capacity was significantly lower in leptin-resistant fetuses (p=0.003), whilst systemic oxidative stress was increased (p=0.02). CONCLUSION: Our results indicate key differences in the antioxidant status in pregnancies affected by leptin resistance.

Polyphenols and IUGR pregnancies: Maternal hydroxytyrosol supplementation improves prenatal and early-postnatal growth and metabolism of the offspring.

Hydroxytyrosol is a polyphenol with antioxidant, metabolism-regulatory, anti-inflammatory and immuno-modulatory properties. The present study aimed to determine whether supplementing the maternal diet with hydroxytyrosol during pregnancy can improve pre- and early post-natal developmental patterns and metabolic traits of the offspring. Experiment was performed in Iberian sows fed a restricted diet in order to increase the risk of IUGR. Ten sows were treated daily with 1.5 mg of hydroxytyrosol per kg of feed between Day 35 of pregnancy (30% of total gestational period) until delivery whilst 10 animals were left untreated as controls. Number and weight of offspring were assessed at birth, on post-natal Day 15 and at weaning (25 days-old). At weaning, body composition and plasma indexes of glucose and lipids were measured. Treatment with hydroxytyrosol was associated with higher mean birth weight, lower incidence of piglets with low birth weight. Afterwards, during the lactation period, piglets in the treated group showed a higher body-weight than control piglets; such effects were even stronger in the most prolific litters. These results suggest that maternal supplementation with hydroxytyrosol may improve pre- and early post-natal development of offspring in pregnancies at risk of IUGR.

Intake of high saturated-fat diets disturbs steroidogenesis, lipid metabolism and development of obese-swine conceptuses from early-pregnancy stages.

The current study indicates that life-long intake, from early-life, of an obesogenic diet with high saturated-fat (HSF) content induces dyslipidemia (high plasma concentrations of triglycerides in concurrence with low concentrations of HDL-cholesterol) in obese swine with leptin resistance (Iberian sows). In case of pregnancy, ovarian features (ovulatory efficiency and luteal steroidogenesis) of sows fed with HSF are not affected but embryo features are affected at so early stages like 28 days of pregnancy (first quarter), although embryo viability was still not affected. In this way, offspring from HSF sows showed a higher incidence of alterations in their developmental trajectory, mainly due to a higher incidence of growth retardation, in their steroidogenic activity and in their availability of triglycerides and cholesterol. In conclusion, the results obtained in the present study illustrate the deleterious effects of maternal dyslipidemia, induced by the intake of HSF diets, on the oestradiol secretion of the conceptuses at early-pregnancy stages and, thus, on their developmental and metabolic features. This article is part of a Special Issue entitled 'Pregnancy and steroids'.

Effect of an obesogenic diet during the juvenile period on growth pattern, fatness and metabolic, cardiovascular and reproductive features of Swine with obesity/leptin resistance.

The objective of this study was to determine, in a female swine model of leptin resistance (Iberian pig), the effect of an obesogenic diet, with high saturated fat content, during the juvenile period, on the appearance of early obesity and its possible effects on metabolic syndrome-related parameters and reproductive features (puberty attainment). Thus, from 130 to 245 days-old, seven Iberian gilts had ad libitum access to food enriched with saturated fat whilst six females acted as controls and had ad libitum access to a commercial maintenance diet. Results showed that a high fat intake-level during the juvenile period induces early obesity with lower body weight and size but a higher body fat-content. Such obesity was related with impairments of glucose regulation predisposing for insulin resistance, but also with an earlier onset of puberty. However, there were no signs of hypertriglyceridemia and hypertension; the gilts diminish their intake level and modify their metabolic features by increasing insulin secretion. In conclusion, Iberian gilts freely eating saturated fat diets during the juvenile period have the prodrome of metabolic syndrome but, during their juvenile period, are still able to develop an adaptive response to the diet.

Characterization of a distinctive pattern of periovulatory leptin secretion and its relationship with ovulation rate and luteal function in swine with obesity/leptin resistance.

Patterns of leptin secretion during the estrous cycle and the possible relationship of changes in circulating leptin during the periovulatory period with ovarian function in sows of obese (Iberian breed) and lean genotype (Large White x Landrace) were evaluated in two consecutive experiments. Plasma leptin concentrations throughout the estrous cycle in lean sows remain unchanged, but Iberian females showed a periovulatory increase in circulating leptin levels without associated changes in body condition and fatness. In these sows, plasma leptin concentrations at Days -1 and 0 of the cycle were found to be positively correlated with the ovulation rate (r=0.943 and r=0.987, respectively; P<0.05 for both), but the levels of leptin at Day 0 were negatively correlated with the progesterone release from Day 3 (r=-0.557; P<0.05) and, became more evident at Day 5 of the estrous cycle (r=-0.924; P<0.005). Such relationships were not observed in the females of the lean genotype. In conclusion, the present study indicates the existence of a distinctive pattern in the periovulatory leptin secretion in swine with obesity and leptin resistance, which is associated with the number and functionality of the corpora lutea present in the subsequent cycle.

The effects of age and reproductive status on blood parameters of carbohydrate and lipid metabolism in Iberian obese sows.

The effects of age and reproductive status on carbohydrate and lipid metabolism of Iberian sows reared in an intensive management system were examined. These animals, with age, are predisposed to hyperglycaemia and dyslipidemia which may develop in response to nutritional challenges during lactation. At weaning, high levels of glucose, triglycerides and low-density lipoproteins cholesterol (LDL-c) with low high-density lipoproteins cholesterol (HDL-c) are indicative of insulin resistance.

Developmental competence of antral follicles and their oocytes after gonadotrophin treatment of sows with gene polymorphisms for leptin and melanocortin receptors (Iberian pig).

PURPOSE: To evaluate possible differences in follicle and oocyte developmental competence after gonadotrophin treatment in sows of obese and lean genotypes. METHODS: Follicle dynamics, ovulation rate and oocyte developmental competence to embryo were compared between females, of obese (n = 7) and lean genotypes (n = 10), treated with 1,250 I.U. of eCG and 500 I.U. of hCG. RESULTS: The obese genotype showed lower numbers of follicles growing to preovulatory stages (12.4 ± 1.8 vs 18.6 ± 1.0, P < 0.05), of corpora lutea (16.0 ± 0.9 vs 23.5 ± 0.9, P < 0.05), and of recovered oocytes/embryos (8.0 ± 1.3 vs 12.9 ± 0.9, P < 0.05). Thereafter, embryo viability rates also decreased when compared to lean genotypes (62.5 vs 77.6%, P < 0.05). DISCUSSION: To our knowledge, this is the first study analyzing the effect of obese genotypes on the ovarian response to exogenous gonadotrophins in a non-rodent animal model, the pig. A lower efficiency of gonadotrophin treatments for stimulation of follicle development and induction of ovulation was observed.

Ovarian response in sheep superovulated after pretreatment with growth hormone and GnRH antagonists is weakened by failures in oocyte maturation.

The administration of growth hormone (GH) or GH plus GnRH antagonists (GnRHa) in sheep allows the enhancement of the pool of gonadotrophin-responsive follicles present in the ovaries and may be useful to increase yields obtained in embryo programmes. The objective of the current study was to evaluate the ability of follicles recruited in response to treatment with GH and GnRHa to grow in response to exogenous follicle stimulating hormone (FSH) and the competence of their oocytes to resume meiosis. Seven females were treated with two doses of GnRHa (days 0 and 3) and three doses of 15 mg of GH (days 3, 4 and 5). Thereafter, this group and a second group (n = 7) were treated with three doses of 1.5 ml of FSH 12 h apart. A third group (control; n = 4) did not receive GH/GnRHa or FSH. The mean number of follicles aspirated on day 7 was higher in ewes treated with GH and GnRHa prior to the stimulation with exogenous FSH than in ewes treated with FSH without pretreatment and in untreated control sheep (20.4 +/- 2.6 vs 17.7 +/- 3.9 and 11.5 +/- 0.8, p < 0.05 and p < 0.01, respectively). The number of recovered cumulus-oocyte complexes after follicular aspiration was higher in the GH/GnRHa + FSH group (8.7 +/- 0.9 vs 6.8 +/- 1.3 in FSH group, n.s., and 4.5 +/- 0.8 in control, p < 0.05), but there were no differences found in the resumption of meiosis (63.1 +/- 9.5% for GH/GnRHa + FSH vs 79.5 +/- 6.3% for FSH and 60.0 +/- 8.8% for control). These results indicate that GH and GnRHa would be useful to increase the number of gonadotrophin-responsive follicles in the ovary, but adjustment of later FSH treatment allowing further development of follicles may be necessary prior to its use in superovulatory protocols.