Regina Célia Teixeira Gomes

Concentration and distribution of hyaluronic acid in mouse uterus throughout the estrous cycle

OBJECTIVE To quantify and study the immunoexpression of hyaluronic acid (HA) in the uterine horns of the mouse throughout the estrous cycle phases. DESIGN Experimental study using an ELISA-like fluorometric assay to quantify HA and an avidin-biotin immunoperoxidase method using biotinylated hyaluronan-binding protein for histochemical studies. SETTING University-based laboratory. ANIMAL(S) Forty regularly cycling adult female mice were divided into four groups according to the diagnosed phase of the cycle: proestrus, estrus, metaestrus, and diestrus. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Histologic samples of the uterine horns. Immunohistochemical reaction was evaluated by detection of HA and CD44 distribution within the uterine horn. Tissue HA content was determined through an ELISA-like fluorometric assay with the same hyaluronan-binding protein and with europium-labeled streptavidin. RESULT(S) The immunohistochemical HA and CD44 reactions were most intense during diestrus, mainly below the luminal epithelium. HA was strongly stained in the connective tissue near the myometrium layer during metaestrus. The biochemical data showed that the highest concentration of HA in uterine horns occurred during diestrus (4053.0 +/- 651.4 ng/g dry tissue) compared with other phases. CONCLUSION(S) Our data show that the expression of HA in mouse uterine horns is highest during the diestrous phase. The fluctuations of HA in the mouse uterus during the estrous phase may be related to the varying estrogen and P levels during the cycle and may be important as far as embryo implantation is concerned.

Hyaluronic acid concentration in postmenopausal facial skin after topical estradiol and genistein treatment: a double-blind, randomized clinical trial of efficacy.

ObjectiveThe aim of this work was to compare the effects of estradiol and genistein treatment on hyaluronic acid (HA) concentration in postmenopausal facial skin. MethodsIn this study, 30 postmenopausal women were evaluated in a prospective, randomized, double-blind trial. The volunteers were postmenopausal women treated in the Gynecology Department of the Federal University of São Paulo. The participants were divided into two groups: group E, treated with 0.01% 17&bgr;-estradiol gel (n = 15), and group G, treated with 4% genistein gel (isoflavones, n = 15). The treatment lasted for 24 consecutive weeks. Preauricular skin biopsies were performed for each participant at baseline (E1 and G1) and after treatment (E2 and G2) to evaluate HA concentration in tissue. The materials were processed using immunohistochemical and biochemical methods. ResultsAfter 24 weeks of treatment, HA concentration increased in both groups, but the effect was greater for estradiol treatment than for genistein treatment. ConclusionsOur data suggest that both treatments may enhance HA concentration in postmenopausal skin but that estrogen produces results that are greater than those produced by isoflavones.

Perfil de glicosaminoglicanos sulfatados no útero de camundongas durante o ciclo estral

OBJECTIVE: Identification and quantitation of sulphated glycosaminoglycans (GAGs) in the uterus of female mice during the estrous cycle. METHODS: Four groups (n = 10 each) of virgin, 100-day old female mice were assembled according to the estrous cycle phase: proestrus, estrus, metaestrus and diestrus. Samples of the median portion of uterine horns were processed for light microscopy examination (H/E and Alcian blue + PAS). The GAGs were extracted and characterized by agarose gel electrophoresis. Data were analyzed by the unpaired Students t-test. RESULTS: At light microscopy GAGs appear in all layers of the uterus, especially in the endometrium, between collagen fibers, in the basal membrane and around fibroblasts. Biochemical analyses disclosed presence of dermatan sulphate (DS), chondroitin sulphate (CS and heparan sulphate (HS) during all estral cycle phases. There was no clear electrophoretic separation between DS and CS, thus these two GAGs were considered together (DS+CS) (proestrus = 0.854 ± 0.192; estrus = 1.073 ± 0.254; metaestrus = 1.003 ± 0.255; diestrus = 0.632 ± 0.443 µg/mg). HS was as follows: proestrus = 0.092 ± 0.097; estrus = 0.180 ± 0.141; metaestrus = 0.091 ± 0.046; diestrus = 0.233 ± 0.147 µg/mg. The uterine content of DS+CS peaked at estrus (estrogenic action) and that of HS at diestrus (progestagen action). CONCLUSION: Due to a constant turnover process, there are definite alterations in the uterine profile of GAGs content during the estrous cycle in mice, which may be modulated by female sex hormones.

Effects of metoclopramide-induced hyperprolactinemia on the prolactin receptor of murine endometrium.

OBJECTIVE To evaluate the effects of metoclopramide-induced hyperprolactinemia on the prolactin receptor of murine endometrium. DESIGN Experimental study using the RNA extraction to detect tissue prolactin receptor isoforms by reverse-transcriptase polymerase chain reaction (RT-PCR). SETTING University-based laboratory. ANIMAL(S) Seventy-two female swiss albino mice (Mus musculus), approximately 100 days old, were divided into six 12-animal groups: (GI) nonoophorectomized mice given vehicle; (GII) nonoophorectomized mice treated with metoclopramide; (GIII) oophorectomized mice treated with metoclopramide; (GIV) oophorectomized mice treated with metoclopramide and 17beta-estradiol; (GV) oophorectomized mice treated with metoclopramide and micronized progesterone; (GVI) oophorectomized mice treated with metoclopramide and a solution of 17beta-estradiol and micronized progesterone. INTERVENTION(S) Drugs were administered for 50 days. Following euthanasia, the middle portions of the uterine horns were removed, sectioned, and immediately frozen for RT-PCR procedures. Blood was collected for the dosage of prolactin and serum estrogen and progesterone using radioimmune assay. MAIN OUTCOME MEASURE(S) Identification of uterine prolactin receptor isoforms. RESULT(S) The PRL receptor and its isoform L were identified only in GI (control group) and GII (metoclopramide), the two groups with nonoophorectomized animals. The amount of PRL receptor mRNA and that of its isoform L from GII were the largest. No other isoforms of the prolactin receptor were identified in any of the groups. CONCLUSION(S) Our results suggest that replacement of estrogen and progestin may not increase the mRNA of endometrial PRL receptor in metoclopromide-induced hyperprolactinemia in rats after castration.

Effects of metoclopramide on the mouse anterior pituitary during the estrous cycle

The symptoms of hyperprolactinemia in women mainly result from changes in the release of gonadotropins and the consequent repercussions on ovarian function.1-3 Metoclopramide is a hyperprolactinemic drug that is used as an antiemetic agent.4,5 In female mice, metoclopramide-induced hyperprolactinemia causes changes in the reproductive system, mainly in ovarian function and the endometrium.6-10 While there is extensive data regarding prolactin regulation and hyperprolactinemic states, the direct effects of metoclopramide on the morphological and functional aspects of pituitary cells remain unknown. Therefore we have evaluated the histomorphological and immunohistochemical changes in lactotrophs of female mice upon treatment with metoclopramide.

The evaluation of endometrial sulfate glycosaminoglycans in women with polycystic ovary syndrome

Abstract The aim of this study was to quantify the sulfated glycosaminoglycans in the endometria of women with polycystic ovary syndrome (PCOS). Of the 18 patients recruited for this study, 10 patients with PCOS comprised the PCOS group (PCOSG), and eight patients with regular and ovulatory menstrual cycles comprised the control group (CG). The clinical, biochemical, morphological and endometrial data from both groups were analyzed. Biopsies were performed during the proliferative phase of the menstrual cycle for the CG and during the persistent proliferative phase for the PCOSG (all women were amenorrheic). In the PCOSG, there was a significant increase in the endometrial concentration levels of heparan sulfate (p = 0.03), but no difference in the concentrations of chondroitin sulfate was determined between the two groups (p = 0.77). Period of time without menstruation (p = 0.001) and body mass index (BMI) (p = 0.04) correlated directly and positively with heparan sulfate concentration. There was no association between heparan sulfate levels and basal insulin values (p = 0.08). High levels of endometrial heparan sulfate in women with PCOS indicate an interference with maternal–fetal recognition, which contributes to infertility; thus, endometrial heparan sulfate may be a predictive marker of future neoplasia risk.

Efeitos da hiperprolactinemia sobre o útero de camundongos no proestro

PURPOSE To evaluate the effect of hyperprolactinemia induced by metoclopramide on the endometrium and myometrium of female mice in the proestrus phase. METHODS 24 female mice were randomly divided in two groups: CtrG/control and ExpG/treated with metoclopramide (6.7 mg/g daily). After 50 days, the animals were sacrificed in the proestrus phase, and the blood was collected to determine the levels of estradiol, progesterone and prolactin. The uterine horns were removed, fixed in 10% formaldehyde and processed before being included in paraffin. Slices of 4 microm were stained by hematoxylin and eosin (H/E). In the morphological analysis, a Carl Zeiss light microscope, with objectives varying from 4 to 400 X was used for each histological slice characterization. In the morphometrical analysis, the superficial epithelium, the lamina propria and the myometrium thickness were evaluated, with the help of an image analyzer (AxionVision - Carl Zeiss) attached to the light microscope (Carl Zeiss). The statistical analysis was done by ANOVA, followed by the Wilcoxon test. P-value was considered as significant, when <0.05. RESULTS Our findings have shown an increase in the seric levels of prolactin (295.6+/-38.0 ng/mL) and significant decrease in the progesterone levels (11.3+/-0.9 ng/mL) in the ExpG, as compared to the CtrG (45.5+/-5.2 ng/mL and 18.2+/-1.6 ng/mL, respectively; p<0.001). Concerning the seric level of estradiol, significant differences between the groups were not obtained (ExpG=119.1+/-12.3 pg/mL and CtrG=122.7+/-8.4 pg/mL; p=0.418). The morphological study has shown that the uterus from the ExpG presented the endometrium with more developed superficial epithelium and lamina propria, as compared to the CtrG, the same happening with the myometrium. The thickness morphometrical values of the luminal epithelium (8.0+/-1.1 microm) and endometrium (116.2+/-21.1x10(2) microm) from the CtrG were lower than the ones from the ExpG (10.2+/-0.8 microm and 163.2+/-23.3x10(2) microm, respectively) with p<0.05. Nevertheless, data obtained in the myometrium have not shown significant differences between the groups (CtrG=152.2+/-25.2x10(2) microm and ExpG=140.8+/-18.0x10(2) microm). CONCLUSIONS Data have shown that hyperprolactinemia induced by metoclopramide determines endometrial proliferation and interferes with the ovarian function, mainly in the progesterone production.

Concentration of glycosaminoglycan in ovariectomized mice uterus after treatment with ovarian steroids.

Abstract The aim of this study was to evaluate the amount of non- and sulfated glycosaminoglycans in the ovariectomized mice uterus, after treatment with ovarian steroids. For this purpose, 50 adult female mice were divided into five groups with 10 animals/each: control group: CG (ovary intact), and ovariectomized groups: OG (vehicle), EG (estradiol), PG (progesterone) and EPG (estradiol combined to progesterone). The treatments started 30 days after ovariectomy. All the animals were treated for 50 consecutive days. These hormones were administered in a sterile oily solution via gavage. Twenty-four hours after the last treatment, all animals were euthanized, removing the uterine horn for biochemical analyses. To quantify, the hyaluronic acid (HA) used ELISA-like fluorometric assay, and the sulfated glycosaminoglycans (GAGs) used agarose gel electrophoresis. The amount of HA was significantly higher in the group treated with progesterone (PG) compared to the others groups (p < 0.05), and in the group treated with estradiol (EG), the amount of chondroitin/dermatan sulfate was significantly higher compared to the others groups (p < 0.05), and in the group treated with progesterone (PG), the amount of heparan sulfate was significantly lower compared to the others groups, except to control group (p < 0.05). Our results showed that the estroprogestative therapy after long time (50 days) profoundly affected the amount of glycosaminoglycans in uterine. These changes may be indicative of uterine pathology such as the development of tumor.

Molecular features of sexual steroids on cartilage and bone

In Brazil, the increase in the reported cases of degenerative diseases of articular cartilage is 20% per year, meaning that 200,000 Brazilians develop degenerative joint diseases every year, which have a negative impact on bone mass. This study shows evidence that hormone production of sexual steroids (estrogens, progestogens, and androgens) have an influence on cartilage quality, as well as on bone mass. Therefore, this review aimed to analyze literature data on the molecular and genic action of sexual steroids on hyaline cartilage and bone physiology, as well as osteoarthritis interference on the quality of these structures.

Aspectos moleculares dos esteroides sexuais sobre a cartilagem e os ossos

In Brazil, the increase in the reported cases of degenerative diseases of articular cartilage is 20% per year, meaning that 200,000 Brazilians develop degenerative joint diseases every year, which have a negative impact on bone mass. This study shows evidence that hormone production of sexual steroids (estrogens, progestogens, and androgens) have an influence on cartilage quality, as well as on bone mass. Therefore, this review aimed to analyze literature data on the molecular and genic action of sexual steroids on hyaline cartilage and bone physiology, as well as osteoarthritis interference on the quality of these structures.