Tatiane da Silva Faria

Effects of high-fat diet on plasma lipids, adiposity, and inflammatory markers in ovariectomized C57BL/6 mice

OBJECTIVE We hypothesized that a high-fat (HF) diet aggravates ovariectomy-related complications. To test this hypothesis, ovariectomized (OVX) mice were fed a HF diet, and we investigated the lipid metabolism, adipose tissue remodeling, adipokines, and inflammatory cytokines. METHODS To investigate the situation in a mouse model of ovariectomy, OVX and SHAM C57BL/6 mice fed a HF diet (60% fat) or standard chow (SC, 10% fat) were monitored for 18 wk. We evaluated daily food intake and weekly body weight. Mice were killed at 30 wk of age. Blood samples and adipose tissue were collected for biochemical, histologic, and molecular analysis. RESULTS OVX groups showed atrophied uterus compared to the SHAM groups, ensuring the success of surgically induced menopause. Despite lower food intake, OVX-HF mice gained about 52% more weight and had heavier total body fats, especially in relation to ovarian fat pad (372%)-a visceral fat which is associated with increased pathogenicity in obesity, and showed larger adipocytes (30%) when compared to OVX-SC mice. Biochemical analysis showed that the OVX-HF mice had increased levels of serum total cholesterol (51%), greater serum triglycerides (158%), lower serum adiponectin (40%), and higher plasma leptin (323%) than OVX-SC mice. The obese group (OVX-HF) also had higher IL-6 levels than both SHAM-HF (241%) and OVX-SC mice (870%). CONCLUSION OVX C57BL/6 mice fed HF diet had greater adipose fat pad, larger adipocytes, and increased inflammatory markers, reinforcing the idea that a HF diet aggravates the complications of ovariectomy-associated inflammation.

Maternal malnutrition during lactation alters the folliculogenesis and gonadotropins and estrogen isoforms ovarian receptors in the offspring at puberty

In this study, we aimed to evaluate whether maternal malnutrition during lactation alters the folliculogenesis and the expression of the gonadotropins and estrogen isoforms ovarian receptors in the offspring at puberty. At parturition, dams were randomly assigned to the following groups: control (C) group, with free access to a standard laboratory diet containing 23% protein and protein-energy-restricted (PER) group, with free access to an isoenergy and protein-restricted diet containing 8% protein. After weaning, the female pups had free access to standard laboratory diet. The maternal malnutrition caused a significant increase in the number of preantral (C=13.72+/-2.87; PER=26.36+/-3.03, P<0.01) and small antral follicles (C=9.32+/-1.35; PER=17.64+/-2.33, P<0.01) and decrease in the number of primordial (C=11.72+/-1.37; PER=3.92+/-0.60, P<0.01) and Graafian follicles (C=1.84+/-0.21; PER=0.96+/-0.11, P<0.01), and corpus luteum (C=2.00+/-0.28; PER=0.80+/-0.31, P<0.01). The estradiol serum concentration was significantly higher (C=67.86+/-4.39; PER=83.29+/-2.68, P<0.05) while testosterone serum concentration did not show statistical difference (C=0.09+/-0.02; PER=0.11+/-0.01, P>0.05) in the PER group. In relation to the receptors expression, maternal malnutrition led to a significant increase in the amount of Fshr (C=0.89+/-0.04; PER=1.07+/-0.03, P<0.05) and Lhcqr (C=0.87+/-0.15; PER=1.33+/-0.08, P<0.05) transcripts and a significant decrease in the amount of Ar (C=0.59+/-0.006; PER=0.13+/-0.080, P<0.05), ER alpha (Esr1) (C=3.33+/-0.71; PER=0.74+/-0.50, P<0.05), ER beta 1 (Esr2) (C=1.33+/-0.06; PER=0.49+/-0.36, P<0.05), and ER beta 2 (Esr2) (C=3.28+/-0.60; PER=0.62+/-0.34, P<0.05) transcripts. In conclusion, perinatal maternal malnutrition can directly affect folliculogenesis at puberty probably as a consequence of changes in the ovarian expression of gonadotropins, androgen and estrogens isoforms receptors. Long-term sexual alterations could be expected in this experimental model, since a reduction in the primordial follicle number is observed, which can result in a decrease in the reproductive lifetime and an earlier termination of breeding capacity.

Effects of maternal undernutrition during lactation on estrogen and androgen receptor expressions in rat ovary at puberty

OBJECTIVE The aim of this study was to evaluate the effects of maternal protein and energy-restricted diets during lactation in folliculogenesis and its relations to androgen and estrogen receptors in the offspring at puberty. METHODS At parturition, dams were randomly assigned to a control (C) group, with free access to a standard laboratory diet containing 23% protein; a protein-energy-restricted (PER) group, with free access to an iso-energy and protein-restricted diet containing 8% protein; and an energy-restricted (ER) group, receiving standard laboratory diet in restricted quantities. After weaning, female pups had free access to standard laboratory diet. RESULTS The number of preantral (C 13.72 ± 2.87, PER 26.36 ± 3.03, ER 26.88 ± 2.31, P < 0.05) and small antral (C 9.32 ± 1.32, PER 17.64 ± 2.33, ER 17.04 ± 2.22, P < 0.05) follicles was significantly increased by maternal malnutrition. The number of primordial follicles (C 10.57 ± 1.61, PER 4.30 ± 0.62, ER 6.28 ± 1.30, P < 0.05), Graafian follicles (C 1.04 ± 0.09, PER 0.52 ± 0.10, ER 0.36 ± 0.11, P < 0.01), and corpus luteum (C 4.84 ± 0.62, PER 2.80 ± 0.50, ER 3.24 ± 0.27, P < 0.05) was significantly reduced. Maternal protein- and energy-restricted diets led to a significant decrease in the androgen (C 9815 ± 1015, PER 6071 ± 838.7, ER 5811 ± 699.3, P < 0.05) and estrogen (C 0.79 ± 0.244, PER 0.12 ± 0.035, ER 0.20 ± 0.036, P < 0.05) α-receptors. In growing follicles, androgen receptor was immuno-expressed in granulosa and theca cells. Estrogen receptor-α was mainly expressed in stroma cells. CONCLUSION Our results suggest that maternal protein- and energy-restricted diets during lactation can disturb the follicular development of the offspring, probably by reducing the number of androgen and estrogen receptors in the ovary.

Metabolic programming of ovarian angiogenesis and folliculogenesis by maternal malnutrition during lactation

OBJECTIVE To evaluate whether maternal malnutrition during lactation programs ovarian folliculogenesis and the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) and its receptors KDR, Flt-1, and FGFR. DESIGN Experimental study. SETTING University-based research laboratory. ANIMAL(S) Adult female rats from a urogenital research laboratory. INTERVENTION(S) Six rat dams randomly assigned to the following groups: control group (C), with free access to a standard laboratory diet containing 23% protein; and a protein-energy-restricted group (PER), with free access to an isoenergy and protein-restricted diet containing 8% protein. After weaning, the female pups had free access to the standard laboratory diet until 90 days of age, when they were sacrificed at the proestrum stage. MAIN OUTCOME MEASURE(S) Quantification of ovarian follicles, vessels, and expression of growth factors and their receptors. RESULT(S) Maternal malnutrition during lactation caused a significant reduction in the number of primordial (C = 6.60 +/- 0.24, PER = 5.20 +/- 0.20), primary (C = 5.80 +/- 0.66, PER = 4.00 +/- 0.31), and Graafian follicles/section (C = 2.18 +/- 0.29, PER = 1.08 +/- 0.37), in KDR (C = 0.22 +/- 0.04, PER = 0.09 +/- 0.01), Flt-1 (C = 0.28 +/- 0.05, PER = 0.12 +/- 0.02), and FGFR mRNA expression (C = 0.34 +/- 0.05, PER = 0.13 +/- 0.05) and in the vessel density of follicles (C = 17.26 +/- 2.30, PER = 9.96 +/- 0.97). CONCLUSION(S) Maternal malnutrition during lactation programs the follicular development by a reduction of VEGF and FGF mRNA receptors expression, probably from a direct action on the follicular development or a reduction in vasculature resulting in a decreased delivery of folliculotrophic substances in PER animals.

Maternal malnutrition during lactation affects folliculogenesis, gonadotropins, and leptin receptors in adult rats.

OBJECTIVE The goal of this study was to evaluate if maternal malnutrition during lactation could possibly program folliculogenesis, the ovarian expression of gonadotropins, leptin, and their receptors. METHODS At parturition, dams were randomly assigned to a control group (C), with free access to a standard laboratory diet containing 23% protein, and a protein-energy-restricted group (PER), with free access to an iso-energy and protein-restricted diet containing 8% protein. After weaning, all female pups had free access to the standard laboratory diet until 90 d of age when they were euthanized in the diestrum stage. RESULTS Maternal malnutrition caused decreases in the number of primordial (C 6.60 ± 0.24, PER 5.20 ± 0.20, P = 0.01), primary (C 5.80 ± 0.66, PER 4.00 ± 0.31, P = 0.04), and Graafian (C 2.18 ± 0.29, PER 1.08 ± 0.37, P = 0.05) follicle numbers. Maternal malnutrition led to a significant decrease in the aromatase mRNA expression (C 0.536 ± 0.008, PER 0.353 ± 0.041, P = 0.01) follicle-stimulating hormone receptor (C 1.25 ± 0.17, PER 0.75 ± 0.02, P = 0.04), luteinizing hormone receptor (C 0.93 ± 0.09, PER 0.54 ± 0.10, P = 0.03), leptin (C 0.55 ± 0.03, PER 0.42 ± 0.03, P = 0.04), Ob-R (C 1.05 ± 0.12, PER 0.64 ± 0.07, P = 0.03), and Ob-Rb (C 1.34 ± 0.21, PER 0.47 ± 0.10, P = 0.02) transcripts when compared with C. CONCLUSION Maternal malnutrition during lactation modulates folliculogenesis and the expression of the different isoforms of leptin and gonadotropin receptors and the aromatase enzyme. This probably is a consequence of alterations in perinatal leptin concentrations that may play a crucial role in determining the occurrence of long-term metabolic changes.

Maternal protein-energy and energy-restricted diets during lactation possibly could program folliculogenesis and the ovarian expression of leptin and its different isoform receptors in rats

Both protein-energy and energy-restricted diets during lactation program the ovarian function of the rat offspring, leading to a reduction of folliculogenesis, possibly as the consequence of the altered expression of leptin and its isoform receptor genes.

Quantitative Morphology Update: Image Analysis

La morfologia cuantitativa es una herramienta confiable en la biologia del desarrollo, clinica y en el envejecimiento. Los innumerables datos cuantitativos de mejoras histologica y / o deterioro debido a las manipulaciones dieteticas o intervenciones farmacologicas atraen la atencion de los investigadores en todo el mundo. La morfometria permite realizar una amplia gama de analisis en dos dimensiones, mientras que la segmentacion de imagenes permite la medicion de una unica estructura o la determinacion de la intensidad de su color despues de una adecuada inmunotincion o incluso el area ocupada por un tipo especifico de celula. Cuando se trata de un analisis imparcial, debe considerarse la posibilidad de la perfusion tisular adecuada, fijacion e inclusion, evitando artefactos. Ademas, se debe considerar la contraccion del tejido. Con el fin de asegurar la reproducibilidad, uno de los principios mas importantes en la investigacion cientifica, y para permitir que diferentes grupos se comparen, la adquisicion y segmentacion de la imagen aparecen como pasos cruciales y deben ser estandarizados. En este trabajo se abordan algunos cuidados importantes para el procesamiento de tejidos y evaluacion apropiadas, y se muestran algunos ejemplos practicos de como la segmentacion morfometrica y la imagen se pueden aplicar a su experimento.

The Effect of Maternal Malnutrition During Lactation on the Endometrial ERα Expression, Collagen Type, and Blood Vessels in the Rats Offspring at Puberty

The aim of this manuscript was to evaluate the effects of maternal protein‐energy‐restriction and energy restriction during lactation on endometrial collagen and blood vessels, uterus Erα expression, and estradiol serum levels in the rats offspring at puberty. At parturition, dams were grouped as: control group (C), with free access to standard rat chow containing 23% protein and 17,038.7 KJ/Kg; protein‐energy restricted group (PER), with free access to formulated chow containing 8% protein but made isoenergetic to the C diet (17,038.7 KJ/Kg); and energy‐restricted group (ER), which received standard rat chow containing 23% protein based on the mean ingestion of the PER group corresponding to 60% of that consumed by the control group. After weaning, all female pups had free access to standard laboratory chow until puberty, when they were killed at the diestrum stage. The uterine ERα expression was determined by Western‐Blot and estradiol serum levels by radioimmunoassay. Endometrial collagen and blood vessels were quantified by stereology. The volumetric density of blood vessels (C = 70.7 ± 2.2; PER = 29.2 ± 2.4; ER = 32.3 ± 3.6; P < 0.001) and endometrial collagen (C = 31.1 ± 1; PER = 26.9 ± 1.0; ER = 26.5 ± 0.7; P < 0.05) were significantly reduced in both malnourished groups. The ER group presented higher estradiol serum levels (C = 69.2 ± 6.4; PER = 73.4 ± 5.5; ER = 101.0 ± 5.4; P < 0.01) in relation to C and PER groups. ERα expression was greater in both malnourished groups (C = 0.11 ± 0.02; PER = 0.41 ± 0.12; ER = 0.35 ± 0.03; P < 0.05). In conclusion, maternal malnutrition during lactation caused changes in endometrial angiogenesis, collagen deposition, and Erα expression in female offspring that will appear in puberty and could affect the reproductive biology of the female offspring. Anat Rec, 2010.

Morphological modification of female bladder after prolonged use of soy-based diets.

OBJECTIVES The aim of this study was to compare the effects of a prolonged use of organic and transgenic soy upon the lipid profile and the collagen/muscle ratio of the detrusor muscle of the bladder. METHODS Wistar rats were fed three different diets from weaning until sacrifice (15 months old): control group (CG) casein-based diet; organic soy group (OSG) organic soy-based diet; genetically modified soy group (GMSG) transgenic soy-based diet. RESULTS There was no difference in the food consumption or in the diet isoflavone components among the groups. Comparing to CG, both OSG and GMSG groups presented a significant (p<0.05) reduction in the body weight, triglycerides, cholesterol and the smooth muscle of the detrusor and a significant (p<0.05) increase of collagen fibers number of the detrusor muscle. CONCLUSIONS These findings call into question that, the prolonged use of soy-based diets can be deleterious to the bladder by altering the collagen/muscle ratio what can cause bladder dysfunctions similar with that occurring during menopause.

The testis of the mice C57/BL6 offspring in adulthood have alterations due to maternal caffeine consumption

PURPOSE To investigate the effects of the maternal caffeine consumption during pregnancy to adult male testis mice offspring. METHODS Twenty pregnant mice were divided into control group (c) and caffeine group (cf). dams received daily saline or 20 mg/kg of caffeine subcutaneously. Male offspring were monitored daily until 13th week. The testis were used to evaluate both the proliferation (pcna) and apoptosis (bax); leptin receptor (ob-r); aromatase; follicle stimulating hormone (fshr), luteinizing hormone (lhr) and androgen receptors (ar); steroidogenic acute regulatory protein (star); vascular endothelial growth factor (vegf) and estrogen receptors (erα and erβ) by western blotting. Serum concentrations of testosterone, estradiol and leptin were measured. RESULTS There was a significant reduction in food intake and the body mass gain (p<0.05) in the cf ; pcna (p=0.01), fshr (p=0.02), star (p=0.0007), vegf (p=0.009), ar (p=0.03) in the cf. while an increase were note in bax (p=0.01), ob-r (p=0.02), lhr (p=0.04) and in the aromatase (p=0.03) in the cf. only erα and erβ were not changed by maternal caffeine. The serum testosterone levels in the cf offspring were 90% lower than in the c offspring (p=0.04). CONCLUSION Maternal caffeine consumption has a role and alters the testis of the offspring in adulthood.