Histochemical and morphometric analysis of endometriotic lesions induced in female rats and treated with dexamethasone

INTRODUCTION: The literature reports that endometrial implants have receptors for steroid hormones primarily stimulated by estrogen and that some treatment strategies have been proposed in experimental models such as the use of synthetic glucocorticoids, for example, dexamethasone. OBJECTIVE: to analyze histochemically and morphometrically endometriotic lesions induced in rats and treated with dexamethasone (0.8 mg/kg/day). MATERIAL AND METHODS: Forty albino female rats (Wistar strain), with 90 days of age, weighing approximately 150 g, were induced with endometriosis and divided into groups: I - rats with endometriosis and evaluated after 34 days, II - rats with endometriosis and evaluated after 47 days, III - rats with endometriosis and 21 days post-surgery treated with dexamethasone for 13 days and IV - rats with endometriosis and 21 days post-surgery treated with dexamethasone for 13 days and euthanized after a period of 13 days starting from the end of treatment. The fragments of endometrial implants were fixed in Bouin, embedded in Paraplast and stained with hematoxylin-eosin and Mallory trichrome. The mean number of glands was compared through nonparametric Tukey-Kramer test (p < 0,05). RESULTS: Dexamethasone reduced inflammation in the endometrial implants, the collagen content in the stroma and decreased significantly the area occupied by glands (GI - 123.25 ± 6.44ª; IGI - 113 ± 6.27ª; GIII - 81.66 ± 3.05b and GIV - 94 ± 6.24b). CONCLUSION: The applied dexamethasone dosage reduces estrogenic effects in endometrial implants in rats.INTRODUÇÃO: A literatura relata que os implantes endometriais possuem receptores para hormônios esteroides, sendo estimulados principalmente pelo estrógeno, e que algumas estratégicas de tratamento têm sido propostas em modelos experimentais, tais como a utilização de glicocorticoides sintéticos, como a dexametasona. OBJETIVO: analisar histoquímica e morfometricamente lesões endometrióticas induzidas em ratas e tratadas com 0,8 mg/kg/dia de dexametasona. MATERIAL E MÉTODOS: Quarenta ratas albinas (linhagem Wistar) com 90 dias de vida, pesando aproximadamente 150 g, foram induzidas à endometriose e divididas em grupos: 1. ratas com endometriose e avaliadas após 34 dias (G1); 2. ratas com endometriose e avaliadas após 47 dias (G2); 3. ratas com endometriose e, após 21 dias do pós-operatório, tratadas com dexametasona por 13 dias (G3) e 4. ratas com endometriose e, após 21 dias do pós-operatório, tratadas com dexametasona por 13 dias e eutanasiadas após um período de 13 dias, contados a partir do término do tratamento com dexametasona (G4). Os fragmentos dos implantes endometriais foram fixados em Bouin, incluídos em paraplast e corados por hematoxilina-eosina e tricrômico de Mallory. As médias do número de glândulas foram submetidas ao teste não-paramétrico de Tukey-Kramer (p < 0,05). RESULTADOS: A dexametasona reduziu a inflamação nos implantes endometriais, o teor de colágeno no estroma e significativamente a área ocupada pelas glândulas (G1= 123,25 ± 6,44ª; G2= 113 ± 6,27ª; G3= 81,66 ± 3,05b; e G4= 94 ± 6,24b). CONCLUSÃO: A dexametasona, na dosagem utilizada, reduz os efeitos estrogênicos em implantes endometriais em ratas.Universidade Federal de PernambucoUniversidade Federal de São Paulo (UNIFESP)Universidade de São PauloUNIFESPSciEL

Combination of Melatonin and Metformin Hydrochloride for Treatment Polycystic Ovarian in Female Rats

Background: Polycystic ovary syndrome (PCOS) is a gynecological endocrine disorder, results in menstrual abnormalities, androgynism and infertility. In the case of women or others animals with PCOS wishing to treat infertility with the aim of becoming pregnant, the most commonly used is metformin hydrochloride. Recent studies have analyzed the combination of metformin hydrochloride with melatonin in oncological treatment but not to treatment of polycystic ovary syndrome (PCOS). The aim of the present study was to analyze the effectiveness of the combination of metformin hydrochloride and melatonin in the treatment of PCOS to improve the fertility of rats and your hormonal alterations.Materials, Methods & Results: This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Committee on the Ethics of Animal Experiments of the University of Federal Rural of Pernambuco (Permit Number: 23081.009130/2010). A total of 50 albino Wistar rats were used. The animal laboratory of an academic research environment, were randomly separated into five groups consisting of 10 females each. After inducing PCOS, the rats were treated with metformin hydrochloride, and/or melatonin, and the results compared with standard and ultrasound confirmed. The physiological similarities were confirmed by our academic researchers morphological science, and published to the association results of effects syndrome induction through constant lighting in reputable magazine recently. This article was analyzed histological of the implantation sites and ovaries, and the estradiol and progesterone levels on the seventh day of gestation, and the other rats for monitoring pregnancy and morphological identification of possible fetal abnormalities, weight measurement and quantification of offspring. The rats were anaesthetized with intraperitoneal injections of ketamine hydrochloride (80 mg/kg) and xylazine (6 mg/kg) to allow analysis of the reproductive organs. Main outcome measures: The study included histopathology, histochemical and quantitative (of the implantation sites) tests, ultrasound analysis, weight benchmarking and ovarian histology tests, as well as comparison of serum estradiol and progesterone levels, and the morphological assessment of offspring. Results paper shows pharmacological treatment reduced the time needed for pregnancy, increased the plasma progesterone levels, the number and weight of offspring, and reduced plasma estrogen levels and collagen fiber grade, improving blastocyst-endometrium interaction and fetal development.Discussion: Our team of researchers confirmed in a previous paper; in addition, the main experimental model used in research about PCOS in recent years, and considered appropriate combination of the drugs caused a physiological reaction similar to responses identified in healthy rats without induction of the POS control group. However, the clinical and physiological effectiveness of the combination should be further explored, especially with respect to the possible side effects on offspring. The treatment with a combination of metformin hydrochloride and melatonin was more effective against hormonal alterations produced by PCOS, allowing a normalization of biochemical parameters during pregnancy, than monotherapeutic treatment with these drugs. In conclusion, proposed drug combination is a viable option to treatment of polycystic ovary syndrome and improved fetal development. This article allows suggest that further research should be conducted to examine effects associated with these drugs in the treatment of diseases of the female reproductive system experimentally. Only such treatment later in animals and humans suggest

Efeitos da administração exógena de melatonina sobre o fígado da prole de ratas Wistar submetidas à corticoterapia durante a prenhez

A dexametasona vem sendo amplamente utilizada em casos de gestação com risco de prematuridade. Entretanto, doses suprafisiológicas podem acarretar complicações e afetar a embriogênese. A melatonina, por sua vez, apresenta efeitos antagônicos aos glicocorticoides. Assim, avaliou-se a ação da dexametasona administrada durante o período gestacional, e a influência da melatonina sobre os efeitos decorrentes da corticoterapia. Foram utilizadas 20 ratas, divididas em 4 grupos: I - ratas prenhes que receberam solução placebo; II - ratas prenhes que receberam dexametasona (0,8mg/kg); III - ratas prenhes tratadas com melatonina (0,5mg/kg) e IV - ratas prenhes que receberam dexametasona e foram tratadas com melatonina. As amostras foram obtidas da prole com 10, 15 e 20 dias de vida pós-natal. Os resultados mostraram que a dexametasona, administrada a partir do 10º dia do período gestacional, promoveu uma maior taxa de mortalidade, redução acentuada no ganho de peso desses animais, bem como alterações hepáticas histopatológicas e morfométricas, como esteatose, necrose, vacuolização celular, além de diminuição nos grânulos de glicogênio, carboidratos totais e aumento das fibras colágenas teciduais. Entretanto, os animais do grupo IV, apresentaram alterações mais atenuadas, com parâmetros semelhantes aos animais do grupo controle. Assim, conclui-se que a administração da melatonina exógena pode proteger o fígado dos animais ou atenuar alguns efeitos decorrentes da corticoterapia

Insect/Bt-cotton interactions: Are immunological variables and hemocyte ultrastructure in Podisus nigrispinus (Hemiptera: Pentatomidae) affected?

Recent studies have shown that ingestion by the army worm Spodoptera frugiperda of Cry1Ac toxin from Bt cotton promotes histochemical and ultrastructural changes in the digestive cells of the predatory pentatomid bug Podisus nigrispinus. Therefore, mindful of the changes in the midgut of the predator, which represents the first line of defence in this insect, our aim was to test the hypothesis that the consumption of Bt cotton-fed S. frugiperda by P. nigrispinus might lead to alterations in components of the immune system of P. nigrispinus. The Cry1Ac toxin level in the leaves of Bt cotton, nitric oxide, phenoloxidase activity, and total proteins were quantified by ELISA. Total and differential hemocyte counts were evaluated, and hemocyte ultrastructure analysis was undertaken. We found that ingestion of the prey fed daily with approximately 23 ± 0.70 ng g-1 Cry1Ac by wet weight of leaves, and expressed by the Bt cotton, induces small ultrastructural changes in the predator's granulocytes and plasmatocytes. However, these changes did not affect the total number and differential and humoral variables analyzed for the bug's hemocytes. © 2013 Copyright Taylor and Francis Group, LLC

Ultrastructure of the antennal sensilla of Alabama argillacea (Hübner, 1823) (Lepidoptera: Erebidae)

Insects have several types of sensilla, the characterization of which has been fundamental to understanding the mechanisms of sensory perception in different species. This study aimed to describe the ultrastructure of the sensilla present on the antennae of Alabama argillacea (Hübner, 1823) (Lepidoptera: Erebidae), an important pest of cotton (Gossypium hirsutum L.) crops, as well as their possible variation between sexes. To do this, the antennae of males and females of A. argillacea were analyzed using scanning electron microscopy. Sensilla morphometry was assessed using photomicrographs, from which the lengths and basal and apical diameters of sensilla were measured using the ImageJ program. Seven types of sensilla were identified on the antennae of A. argillacea: sensilla trichodea, sensilla chaetica, sensilla auricillica, sensilla coeloconica, sensilla styloconica, sensilla basiconica, and sensilla Böhm bristles. Differences between the sensilla of males and females were found in their lengths and basal diameters in the distal and proximal regions. This suggests that sensilla functionality may not only vary from one species to another, but also between sexes within the same species. Thus, further transmission electron microscopy and single sensillum recording studies may provide more detailed information on the sensilla of A. argillacea and their functions. Keywords: Antenna, Cotton leafworm, Morphometry, Scanning electron microscop

Associative administration of cimetidine and exogenous melatonin on the endometrial receptors of estrogen, collagen and hormone levels in adult rats

Cimetidine hydrochloride, a blocker of H2 receptors in gastric parietal cells, acts by reducing the secretion of acid in the stomach and has been studied as a xenoestrogenic substance. The chronic use of cimetidine produces homonal disorders and toxicity in the male reproductive system, in addition to reducing 2-hydroxylated estradiol and increasing serum levels of estradiol and prolactin in women, leading to hyperprolactinemia, which can be a risk factor for cancer. Melatonin, a neurohormone synthesized by the pineal gland, it has an important role in reproductive function, regulating the production of estrogen, progesterone and prolactin. This study tested the hypothesis that melatonin can block or reduce the estrogenic effects of cimetidine in the uterine stroma interfering in the estrogen receptors, collagen fiber content and hormone levels in adult rats. Forty-five (45) albino female rats were divided into three groups: I – treated with placebo (control); II – treated with cimetidine (50 mg/kg) and III – treated with cimetidine (50 mg/kg) associated with melatonin (200 μg/100g). Experiments were conducted for 7, 14 and 19 days. Cimetidine promoted more intense marks of REα receptors in the uterus of animals treated for 19 days, in the distribution of collagen fibers in the endometrium, elevated serum levels of estrogen and prolactin and progesterone reduction. Melatonin associated with cimetidine has blocked these effects. The melatonin has cytoprotective activity to cimetidine chronic effects on endometrial stroma by reducing or preventing the increase in the synthesis of collagen fibers by fibroblasts by regulating the activity of serum estrogen as well as the expCloridrato de cimetidina, um bloqueador de receptores H2 das células parietais gástricas, que age reduzindo a secreção de ácido no estômago e tem sido estudado como substância xenoestrogênica. O uso crônico da cimetidina produz distúrbios homonais e toxidade no aparelho reprodutor masculino, além de reduzir o estradiol 2-hidroxilado e aumentar níveis séricos de estradiol e prolactina em mulheres, levando a hiperprolactinemia, que pode ser fator de risco para o câncer. A melatonina, neurohormônio sintetizado pela glândula pineal tem importante papel na função reprodutiva, regulando a produção de estrógeno, progesterrona e prolactina. O estudo testou a hipótese de que a melatonina pode bloquear ou reduzir os efeitos estrogênicos da cimetidina no estroma uterino, interferindo nos receptores de estrógeno, no teor de fibras colágenas e nos níveis hormonais em ratas adultas. Quarenta e cinco (45) ratas albinas divididas em três grupos: I – tratadas com placebo (controle); II – tratado com cimetidina (50 mg/kg) e III – tratado com cimetidina (50 mg/kg) associada à melatonina (200 μg/100 g). Os experimentos foram conduzidos por 7, 14 e 19 dias. Nos grupos tratados apenas com cimetidina, observou-se marcações mais intensas dos receptores REα, maior distribuição das fibras colágenas no endométrio, elevação dos níveis séricos de estrogênio, prolactina e redução da progesterona, nos animais tratados por 19 dias. Na associação cimetidina e melatonina, acredita-se que a melatonina bloqueou esses efeitos. A melatonina tem atividade citoprotetora para efeitos crônicos da cimetidina no estroma endometrial, por reduzir ou prevenir o aumento da síntese de fibras de colágeno pelos fibroblastos regulando a atividade do estrogênio sérico, bem como a expressão de seus receptores endometriais, além de manter os níveis normais de progesterona e prolactina.Palavras chave: Melatonina, receptor de estrógeno, xenoestrógeno, morfometria, níveis hormonais