Rejane Mosquette

Estudo morfológico e molecular da mama de ratas castradas tratadas com isoflavona ou estrogênios

PURPOSE: to analyze the effects of isoflavones and estrogens on the morphology, morphometry and VEGF expression of the adult female rat mammary gland. METHODS: Forty-five adult female rats were oophorectomized; 28 days after surgery they were divided into 3 groups of 15 animals each: CON - control (treated with propylenoglycol); ISO - isoflavones (100 mg/kg) and CEE - conjugated equine estrogens (50 µg/Kg). Drugs or vehicle were given orally once a day for 60 days. After this, the animals were killed and the first pair of inguinal mammary glands was immediately removed; part of the material was processed for routine histological study and the remaining tissue was frozen for further analyses of the expression of VEGF mRNA by means of the RT-PCR technique. RESULTS: We observed that mammary ducts were atrophic in the control (CON) and isoflavone-treated (ISO) groups. In these groups the mammary glands were composed of a large concentration of adipose tissue with some ducts and rare alveolar structures. In the CEE group the ducts were well developed with many buds and alveolar structures. The number of mammary gland alveoli was higher in CEE than in the other groups (CON = 1.4 ± 2.1; ISO = 1.6 ± 3.8; CEE = 12.3 ± 7.1 alveoli/mm2; p<0.05%); also, the cell volume was higher (CON = 14.9 ± 4.9; ISO = 11.4 ± 6.9; CEE = 27.4 ± 9.7 µm3, p< 0.05%). The same was observed with regard to the number of blood vessels (CON = 16.4 ± 1.5; ISO = 18.4 ± 2.1; CEE = 37.1 ± 4.1 vessels/mm2, p< 0.05). The expression of VEGF in the CEE group was higher than in the other groups, which did not significantly differ from each other in this respect. CONCLUSION: Our data did not show any proliferation effect in the mammary tissue of adult oophorectomized rats treated with isoflavones (100 mg/kg) during 60 days.

Efeito da hiperprolactinemia induzida pela metoclopramida na glândula lacrimal: estudo experimental

PURPOSE: to evaluate the morphological changes in murine lacrimal glands by metoclopramide-induced hyperprolactinemia during the proestrus phase or pregnancy. METHODS: forty adult mice were divided into two groups: CTR1 (control) and MET1 (treated with metoclopramide). After fifty days, half of the mice were sacrificed. The remaining animals were mated, and then labeled as pregnant controls (CTR2). Part of these animals were treated with metoclopramide and constituted the metoclopramide-treated pregnant (MET2) group. The CTR2 and MET2 groups were sacrificed on the 6th day of pregnancy. The blood was collected for determination of the hormonal levels of estradiol and progesterone by a chemoluminescent method. The lacrimal glands were then removed, fixed in 10% formaldehyde and stained with HE. The morphometric analysis was performed using the Axion Vision program (Carl Zeiss) to measure acinar nuclear and cellular volumes. RESULTS: the nuclear and cellular volumes of the lacrimal glands in the MET1-(152.2±8.7; 6.3±1.6 µm3) and MET2-(278.3±7.9; 27.5±0.9 µm3) treated groups were lower than those in CTR1 (204.2±7.4; 21.9±1.3 µm3) and CTR2 (329.4±2.2; 35.5±2.0 µm3), respectively. There was a significant hormonal level reduction in the animals that received metoclopramide compared to controls (CTR1: estradiol = 156.6±42.2 pg/ml; progesterone = 39.4±5.1 ng/ml; MET1: estradiol = 108.0±33.1 pg/ml; progesterone = 28.0±6.4 ng/ml; CTR2: estradiol = 354.0±56.0 pg/ml; progesterone = 251.0±56.0 ng/ml; MET2: estradiol = 293.0±43.0 pg/ml, progesterone = 184.0±33.0 ng/ml). CONCLUSION: metoclopramide-induced hyperprolactinemia produced morphological signs of reduction of cellular activity in lacrimal glands during the proestrus phase and pregnancy. It is hypothesized that this effect might be related to the hyperprolactinemia-induced decrease in the hormonal production of estrogen and progesterone.

Oxigenoterapia hiperbárica em lesões actínicas de colo de ratos: aspectos morfológicos e morfométricos

Objetivo: Avaliar a acao da oxigenacao hiperbarica em lesoes actinicas provocadas por radiacao ionizante no colo distal de ratas. Metodos: Foram utilizadas 32 ratas adultas distribuidas em quatro grupos. I - sham; II - oxigenoterapia hiperbarica (OHT); III - radioterapia (RT), e IV - radioterapia e oxigenoterapia hiperbarica (da mesma maneira que nos grupos II e III). Segmentos de colo distal foram fixados em formol a 10% e processados para avaliacao histologica e morfometrica. Resultados: Os resultados do experimento mostraram lesao das glândulas intestinais, vasodilatacao e infiltrado leucocitario na lâmina propria e submucosa no colo dos animais pertencentes ao grupo RT que diminuiu acentudadamente apos a oxigenioterapia hiperbarica. Conclusao: A oxigenitoterapia hiperbarica atenua as alteracoes morfologicas da mucosa e submucosa do colon distal de ratas com enterite induzida por radiacao ionizante.

Efeitos das isoflavonas sobre o assoalho pélvico e a vascularização peri-uretral de mulheres na pós-menopausa

PURPOSE: to evaluate muscular strength of the pelvic floor and the periurethral vessels of postmenopausal women before and after six months of soybean extract treatment. METHODS: the study was conducted on 30 postmenopausal women before and after six consecutive months of soyabean extract (100 mg/day) administration. Urinary loss and muscular strength of the pelvic floor were investigated through digital perineometer and functional evaluation. Digital color Doppler in the periurethral region was used to count the number of vessels. For statistical analysis, the paired Student t test was applied to compare the results before and after the treatment. RESULTS: twenty women reported urinary incontinence before the treatment period. The amelioration of this symptom was observed in 15 (75%) women after the treatment. Vaginal pressure (muscular strength of the pelvic floor) was 12.95±1.73 and 15.86±1.86 Sauers, before and after the treatment, respectively (p<0.001). Twenty-two women (73.3%) presented an increase in the pressure at the end of this study. In relation to the function evaluation, 18 (60%) had improvement in muscular strength and 12 women did not present any change. On ultrasonography (Doppler), the number of vessels was 2.20±0.15 blood vessels/field in the beginning of this study and 3.46±0.25 blood vessels/field at the end of the treatment (p<0.001). An increase in the number of periurethral vessels was detected in 21 women (70%). CONCLUSION: it is important to emphasize that these are preliminary results. A double blind randomized and placebo-controlled clinical trial with a high number of participants is necessary. However, the treatment with concentrated soybean extract (100 mg per day) for six consecutive months may determine an improvement in pelvic floor muscular strength and an increase in the number of periurethral vessels in postmenopausal women.

Efeito da hiperprolactinemia induzida pela metoclopramida na córnea de camundongas

PURPOSE: To evaluate the morphological changes in murine cornea upon metoclopramide-induced hyperprolactinemia during the proestrous phase or pregnancy. METHODS: Forty adult mice were divided into two groups: (control) CTR1 and (treated with metoclopramide (MET1). After fifty days, half of the mice were sacrificed. The remaining animals were mated, and then labeled as pregnant controls (CTR2). Part of these animals were treated with metoclopramide and constituted the metoclopramide-treated pregnant (MET2) group. The groups CTR2 and MET2 were sacrificed on the 6th day of pregnancy. The hormonal levels were assessed by chemioluminescence and radioimmunoassay methods and the cornea was removed for the histomorphometric study. RESULTS: The epithelial, stromal, endothelial and total thickness in the experimental group was: MET1 and MET2 were higher than one in the control group: CTR1 and CTR2. There was a significant reduction of the hormonal level in the animals that received metoclopramide as compared to controls (CTR1: estradiol = 156.6±42.2 pg/ml; progesterone = 39.4±5.1 ng/ml; prolactin = 130.4±26.2 ng/ml; MET1: estradiol = 108.0±33.1 pg/ml; progesterone = 28.0±6.4 ng/ml; prolactin = 551.5±23.3 ng/ml; CTR2: estradiol = 354.0±56.0 pg/ml; progesterone = 251.0±56.0 ng/ml; prolactin = 423.2±28.1 ng/ml; MET2: estradiol = 293.0±43.0 pg/ml; progesterone = 184.0±33.0 ng/ml; prolactin = 823.1±51.1 ng/ml). CONCLUSION: The metoclopramide-induced hyperprolactinemia may increase corneal layers, mainly in pregnant mice. Possibly, this effect is related to reduction in estrogen and progesterone production.

Efeitos das isoflavonas sobre o miométrio de ratas adultas

PURPOSE: to evaluate histomorphometric changes in the rat myometrium upon treatment with isoflavones, as compared with estrogens, using histological and morphometric techniques. METHODS: twenty-eight oophorectomized adult rats were randomly divided into four treatment groups: GPropi = propylene glycol (control); GExtr10 - 10 mg/kg soybean extract; GExtr300 - 300 mg/kg soy bean extract; GCee - 200 µg/kg conjugated equine estrogens (Cee). Drugs or drug vehicle were administered by gavage once a day for 21 days. Upon sacrifice, the uteri were removed and weighed. Fragments of uterine horns were collected and fixed in 10% formaldehyde and processed for paraffin inclusion. The histological sections were stained by hematoxylin and eosin and evaluated microscopically by means of an image analyzer to quantify the myometrial thickness and the number of blood vessels and eosinophils. The data were studied by analysis of variance (ANOVA) followed by the Tukey-Kramer multiple comparison test. RESULTS: isoflavones in the concentration of 300 mg/kg induced a significant increase in the myometrium thickness (GExtr300=25.6±5.0 mm) compared to control (GPropi=5.5±0.5 mm). The effect of this high dose is similar to the estrogen effect (GCee=27.5±7.9 mm). In low doses (10 mg/kg), the effect was similar to control. Isoflavones (GExtr300) induced also an increase in the number of blood vessels (GPropi=3.5±1.6; GExtr300=10.2±3.6 vessels/mm2) and of eosinophils (CPropi=0.15±0.01; GExtr300=4.3±0.9 eosinophils/mm2). These effects were comparable to those produced by Cee treatment in GCee (9.2±1.1 eosinophils/mm2). CONCLUSION: a high-dose treatment with isoflavones (300 mg/kg per day, 21 days) elicited an estrogen-like, highly significant proliferative action on the rat myometrium.

Aspectos morfológicos da junção escamo-colunar de ratas em estro permanente e tratadas com associação de estrogênio e glicocorticóide

OBJETIVO: avaliar o efeito da associacao de dexametasona com estrogenio sobre a juncao escamo-colunar (JEC) de ratas em estro permanente e ovariectomizadas (Ovx). METODOS: trinta ratas foram divididas em seis grupos com cinco animais cada: GEF em estro fisiologico, tendo recebido propilenoglicol (veiculo); GOVX em estro fisiologico com ovariectomia e tratadas com o veiculo; GEP em estro permanente (EP); GEPOVX em EP, Ovx e tratadas com veiculo; GESTR - em EP, Ovx e tratadas com 10 mg/dia de benzoato de estradiol e GDEXAR em EP, Ovx e tratadas com 10 mg/dia de benzoato de estradiol e 0,8 mg/dia de dexametasona. O EP foi induzido com 1,25 mg/animal de propionato de testosterona logo apos o nascimento. Apos 90 dias, as ratas foram ovariectomizadas nos grupos GOVX, GEPOVX, GESTR e GDEXAR. Apos 21 dias de castracao, os animais foram tratados por cinco dias consecutivos. No final do experimento, todos os animais foram sacrificados e o utero removido para rotina histologica. RESULTADOS: a JEC do GEP tinha limites irregulares e pouco nitidos, com varios brotamentos em direcao da lâmina propria, bem como reducao do numero de leucocitos comparado a GEF. A JEC do GOVX e do GEPOVX se mostrou pouco nitida, com epitelio cubico na parte endometrial e diminuicao das camadas do epitelio escamoso, com atrofia estromal. No GESTR, a JEC apresentou-se bem mais desenvolvida que nos grupos GOVX e GEPOVX, mas assemelhando-se mais ao GEP devido aos limites pouco nitidos e aumento dos brotamentos. Ja no GDEXAR, a JEC foi bem delimitada, se aproximando do aspecto do GEF. CONCLUSAO: nossos dados sugerem que a dexametasona associada ao estrogenio seria importante na restauracao da morfologia normal da JEC em ratas que foram previamente induzidas a estro permanente e subsequentemente submetidas a castracao.