Ricardo Santos Simões

Efeitos da melatonina no sistema genital feminino: breve revisão

Melatonin is secreted by the pineal gland and this is linked to the day/night cycle. It is an antioxidant and plays a fundamental role in the regulation of the jet-lag stage, in several physiological reactions and in control of the biologic rhythm. Human melatonin has an important influence on the female genital system. In fact, melatonin may influence production and action of steroids, modifying cellular signalization on the target tissue. There are many evidences that the melatonin therapy may be interfering with neoplasia development, mainly of the estrogen-dependent tumor. This paper aims to analyze the actions of melatonin on the neuroendocrine, immunological and cardiovascular systems, as well as on the reproductive function.

Effect of conjugated equine estrogens and tamoxifen administration on thyroid gland histomorphology of the rat

OBJECTIVE The aim of this study was to evaluate the action of the conjugated equine estrogens and tamoxifen on the morphology of thyroid gland in ovariectomized (OVx) rats. METHODS Conjugated equine estrogens (CEE), clinically used as estrogen therapy, is a complex formulation containing multiple estrogens that decrease menopausal symptoms. Thirty ovariectomized rats were randomly divided into 3 treatment groups: GI, vehicle (propylene glycol); GII, CEE 200 microg/kg per day; and GIII, tamoxifen 1 mg/kg per day. Another group of 10 rats with intact ovaries (GIV) was included, treated with the vehicle, and sacrificed during estrous. All animals were treated by gavage for 50 days, after which they were sacrificed. Blood samples were collected, and the thyroid was removed for morphological analysis and PCNA evaluation through immunohistochemical study. RESULTS The thyroid follicular cell height was increased in animals treated with CEE (14.90 +/- 0.20 microm), with TAM (14.90 +/- 0.10 microm), and in rats with intact ovaries (15.10 +/- 0.50 microm) in comparison to that of the vehicle group (9.90 +/- 0.20 microm) (P < 0.001). The follicular area was larger in the CEE (2,225 +/- 51 microm2) and TAM (2,127 +/- 67 microm2) groups compared to that of the vehicle group (5,016 +/- 53 microm2). The levels of T4 and T3 in rats treated with CEE, with Tamoxifen and in rats with intact ovaries, were higher than those those in the vehicle group (P < 0.001). The PCNA index in the vehicle group was lower than in other groups. CONCLUSION Our data suggest that estrogen and tamoxifen administration has a proliferative effect on the thyroid.

Effects of aroeira (Schinus terebinthifoliu Raddi) oil on cutaneous wound healing in rats

PURPOSE To evaluate the effects of aroeira (Schinus terebinthifolius) ointment on skin wound healing in rats. METHODS Adult male rats (n=20) were divided into four groups of five animals each, as follows: G4, G7, G14 and G21, which corresponds to 4th, 7th, 14th and 21th days postoperatively. Each animal were made two incisions on the skin, including the subcutaneous tissue, in the right and left sides of thoracic region, separated by a distance of two inches. The right lesion was treated with base ointment (vaseline, lanolin); the left one was treated with base ointment containing 5% of aroeira oil. At the end of each experimental period the lesions were evaluated for the contraction degree. Then held the collection of fragments that were fixed in 10% formalin and processed for paraffin embedding. In the histological sections (5μm) was evaluated the morphology and quantified the collagen and blood vessels. The data obtained were submitted to ANOVA test complemented by Tukey-Kramer test (p<0.05). RESULTS The contraction of the lesions was higher in wounds treated with aroeira oil than in controls at 7th and 14th days (p<0.01), whereas in the 21st day all lesions were already completely healed. The morphology showed granulation tissue more developed, with fibroblasts more bulky and collagen fibers more arranged in the experimental group at 4th, 7th and 14th days. The morphometry showed a significant increase in the quantification of collagen fibers in the experimental group at 7th and 14th days (p<0.05). CONCLUSION The aroeira oil accelerates the healing process of wounds as a macroscopic, morphological and morphometrical analysis.

Systematic review of cell adhesion molecules and estrogen receptor expression in the endometrium of patients with polycystic ovary syndrome

Infertility associated with polycystic ovary syndrome (PCOS) could be related to many mechanisms including endometrial factors.

Effects of angico extract (Anadenanthera colubrina var. cebil) in cutaneous wound healing in rats

PURPOSE To study the effects of the angico extract (Anadenanthera colubrina var. cebil) on the healing of rat skin. METHODS Twenty adult rats were divided into four groups of five animals each, the G4, G7, G14 and G21, which corresponds to the respective postoperative days. Each group received two incisions on skin and subcutaneous tissue in the right and left antimere of the thoracic region, separated by a distance of 2 cm. The right lesion was treated daily with saline and the left with the angico alcoholic extract (5%). At the end of each experimental period, animals were euthanized and fragments of the wound area, together with the edges were removed, fixed in 10% formaldehyde solution and processed for paraffin embedding. In the histological sections with 5 µm of thickness, were carried out immunohistochemical methods for detection of blood vessels (VEGF) and stained with hematoxylin and eosin for morphological analysis. Statistical analysis was done by ANOVA and Tukey test (p<0.05). RESULTS Morphological analysis showed larger fibroblasts and a higher concentration of collagen fibers in days 7 and 14 in wounds treated with the angico extract. Morphometric analysis demonstrated a significant increase in the number of blood vessels in both the seventh and 14th days (p<0.01) in wounds treated with the angico extract. CONCLUSION The angico alcoholic extract (Anadenanthera colubrina var. cebil) induces the acceleration of wound healing in skin wounds of rats.

Evaluation of FAS and caspase-3 in the endometrial tissue of patients with idiopathic infertility and recurrent pregnancy loss

OBJECTIVE To investigate expression of BcL-2, FAS, FAS ligand (FASL) and cleaved caspase-3 in the endometrial tissue of women with idiopathic infertility (with two consecutive failed cycles of in vitro fertilization) and women with idiopathic recurrent pregnancy loss. The control group consisted of fertile women. STUDY DESIGN Endometrial tissue samples from fertile women (n=25), women with idiopathic infertility (n=25) and women with idiopathic recurrent pregnancy loss (n=25) were collected on the seventh or eighth postovulatory day of their menstrual cycles for evaluation. Expression of BcL-2, FAS, FASL and cleaved caspase-3 was assessed using immunohistochemical methods. RESULTS Expression of BcL-2 and FAS was significantly higher and lower, respectively, in the women with idiopathic infertility than in the other groups (p<0.01). Expression of cleaved caspase-3 was significantly lower in the women with idiopathic infertility than in the other groups (p<0.01). Expression of FASL was similar in all three groups. CONCLUSION Disturbances in endometrial apoptosis may be a contributing factor in patients with idiopathic infertility and recurrent pregnancy loss.

The soybean concentrated extract proliferates the vagina of adult rats

Objective:The aim of this study was to evaluate changes induced on the vagina of ovariectomized rats after treatment with soybean concentrated extract or conjugated equine estrogens and the association of both drugs. Methods:We conducted an experimental study with 50 ovariectomized rats that were randomly divided into five equal groups of 10 animals: GI received vehicle, GII received soybean concentrated extract 46 mg/kg per day, GIII received soybean concentrated extract 120 mg/kg per day, GIV received conjugated equine estrogens 50 &mgr;g/kg per day, and GV received conjugated equine estrogens 50 &mgr;g/kg and soybean concentrated extract 46 mg/kg per day. The substances were administered by gavage during 21 consecutive days. After that, the animals were killed under anesthesia and the vagina was removed for histological and immunohistochemical analysis. Data were initially submitted to analysis of variance. Whenever a significant difference was detected, the study was complemented with the Tukey-Kramer test for multiple comparisons. Results:GII did not show any differences on the vaginal epithelium or collagen compared with GI. GIII presented an increase in vaginal epithelium and collagen amount. GIV had the highest amount of collagen and the signals of vaginal proliferation. GV did not show any additional effect compared with GIV. Conclusions:Our data suggest that a high dose of isoflavone-rich soy extract may have positive effects on the vaginal structures of ovariectomized rats, but this action is less than that of estrogen treatment on vaginal thickness. In addition, soy extract may not block the estrogen effect on vaginal tissue.

Remote ischemic preconditioning on neovascularization and follicle viability on ovary autotransplantation in rats.

PURPOSE Verify the optimum remote vascular occlusion time to reduce ovarian injury in autologous transplants in rats. METHODS Twenty-four adult female rats were assigned to four groups: GC (control group): bilateral oophorectomy followed by ovary transplant; GIPC (ischemic preconditioning group): remote ischemic preconditioning at the iliac artery for 5, 10, and 15 minutes (GIPC-5, GIPC-20, and GIPC-15) previous to bilateral oophorectomy and ovarian transplantation. The right ovary was fixed in the retroperitoneum. Euthanasia was performed 4 days after the surgical procedure. The follicles were counted and classified as developing versus atretic. The immunohistochemical assay identified vascular factor of endothelial growth (VEGF) in the ovarian stroma and assessed the proliferation capacity by means of the Ki-67 in the ovarian follicles. RESULTS Every group showed an inflammatory infiltrate, luteous body, and ovarian follicles in several phases of development. The ischemic preconditioning groups displayed greater amounts of viable ovarian follicles and increased vascularization and vasodilatation than the control group. GIPC-15 showed the highest amount of viable follicles compared to the others (P < .05 GIPC-15 vs GC; GIPC-15 vs GIPC-5). More VEGF-labeled cells were observed in GIPC-10 than the control group (P < .05, GIPC-10 vs GC). The proliferation index assessed by Ki-67 marking showed GC: 80%; GIPC-5: 76%; GIPC-10: 67%; and GIPC-15: 64% (P > .05). CONCLUSIONS The PCI-15 cohort seem to be the most adequate timing to achieve functional support and preservation of a greater number of viable ovarian follicles.

Rat pinealectomy: a modified direct visual approach

PURPOSE To report a new, direct visual approach for rat pinealectomy. METHODS Eighty adult female rats (Rattus norvegicus albinus EPM-1 strain) were weighted and anesthetized intraperitoneally with 15 mg/kg xylazine and 30 mg/kg ketamine. The animal was fastened to a dissection table, an incision was made in the skin and the subcutaneous tissue, bringing the lambda into view. The skullcap was opened with a dental drill, bringing the cerebral hemispheres and the superior sagittal sinus into view. The pineal gland, located under the venous sinus, was removed in a single piece using tweezers. Next, the bone fragment was returned to its place and the surgical layers were sutured. RESULTS This new technique is easy to be done, avoids bleedings and removes only the pineal gland without damage to the remaining encephalon. In addition it makes possible the achievement of a sham surgery, allowing the pineal gland to remain intact. CONCLUSION The proposed technique intends to facilitate studies aiming to better understanding the complexity and importance of the pineal gland on reproductive and other body systems.

Proliferation of the superficial epithelium of ovaries in senile female rats following oral administration of conjugated equine estrogens

OBJECTIVE To evaluate the effect of different concentrations of estrogen on the ovarian superficial epithelium in senile female rats. Design: Fifty female rats at 15 months of age and with irregular estrous cycles were selected and randomly divided into five experimental groups containing equal numbers of animals in each: GPROP, control group receiving vehicle only; GE0.05mg, group receiving conjugated equine estrogens (CEE) at a dose of 50 μg/kg; GE0.5mg, group receiving CEE at 500 μg/kg; GE1mg, group receiving CEE at 1 mg/kg; and GE2mg, receiving CEE at 2 mg/kg. The length of treatment was 21 days. After this period, the animals were anesthetized and the ovaries were fixed in 10% formaldehyde and processed for routine histology. Histomorphology was analyzed by light microscopy, and histomorphometrics were evaluated using the Imagelab program. RESULTS In the GPROP and GE0.05mg groups, the superficial epithelium of the ovary had a simple cuboidal shape, and as the estrogen dose increased, the epithelium thickened, with pseudo-stratified or stratified epithelium appearing in the GE2mg group. The animals in the group given the highest estrogen dose (GE2mg) showed the thickest ovarian epithelium and the largest perimeter and surface area of the surface ovarian epithelium (P < 0.01). However, the difference in epithelium thickness between the GE0.5mg and GE1mg groups was only slight. CONCLUSION Our data suggest that CEE at a dose of 2 mg/kg may induce marked proliferation of rat ovarian epithelium.