RESUMO
Endometriosis is a gynecological disease caused by the growth of endometrial cells outside the uterus leading to inflammation, pelvic pain, and infertility. The relationship between the amount of ectopic uterine tissue growth and the severity of symptoms is still unclear. The presence or degree of pain and infertility does not correlate with the stage of disease as currently defined. Here, we report a clear dose-response relationship between the amount of ectopic tissue transplanted and the reproductive outcomes in a murine model of endometriosis. Endometriosis was induced in mice using various amounts of transplanted uterine tissue. Four groups of mice consisted of a sham surgery control or those transplanted with 1, 2, or 4 endometrial segments of 5 mm each. Pregnancy rates were significantly lower in those transplanted with 2 or 4 segments compared to sham or the 1 segment groups. We demonstrate that infertility does correlate with the extent of active disease. Current clinical staging systems do not account for disease activity and may inappropriately weight sequela of disease. Early recognition and treatment in women may help to minimize the effect of endometriosis on fertility. Here, we describe a mouse model of endometriosis and infertility that may be useful to elucidate the mechanisms of infertility in endometriosis.
Assuntos
Endometriose/fisiopatologia , Resultado da Gravidez , Taxa de Gravidez , Animais , Modelos Animais de Doenças , Endométrio/transplante , Feminino , Camundongos Endogâmicos C57BL , GravidezRESUMO
BACKGROUND: The development and long-term survival of endometriotic lesions is crucially dependent on an adequate vascularization. Hyaluronic acid (HA) through its receptor CD44 has been described to be involved in the process of angiogenesis. OBJECTIVE: To study the effect of HA synthesis inhibition using non-toxic doses of 4-methylumbelliferone (4-MU) on endometriosis-related angiogenesis. MATERIALS AND METHODS: The cytotoxicity of different in vitro doses of 4-MU on endothelial cells was firstly tested by means of a lactate dehydrogenase assay. The anti-angiogenic action of non-cytotoxic doses of 4-MU was then assessed by a rat aortic ring assay. In addition, endometriotic lesions were induced in dorsal skinfold chambers of female BALB/c mice, which were daily treated with an intraperitoneal injection of 0.9% NaCl (vehicle group; n = 6), 20 mg/kg 4-MU (n = 8) or 80 mg/kg 4-MU (n = 7) throughout an observation period of 14 days. The effect of 4-MU on their vascularization, survival and growth were studied by intravital fluorescence microscopy, histology and immunohistochemistry. MAIN RESULTS: Non-cytotoxic doses of 4-MU effectively inhibited vascular sprout formation in the rat aortic ring assay. Endometriotic lesions in dorsal skinfold chambers of 4-MU-treated mice dose-dependently exhibited a significantly smaller vascularized area and lower functional microvessel density when compared to vehicle-treated controls. Histological analyses revealed a downregulation of HA expression in 4-MU-treated lesions. This was associated with a reduced density of CD31-positive microvessels within the lesions. In contrast, numbers of PCNA-positive proliferating and cleaved caspase-3-positive apoptotic cells did not differ between 4-MU-treated and control lesions. CONCLUSIONS: The present study demonstrates for the first time that targeting the synthesis of HA suppresses angiogenesis in developing endometriotic lesions. Further studies have to clarify now whether in the future this anti-angiogenic effect can be used beneficially for the treatment of endometriosis.
Assuntos
Endometriose/etiologia , Ácido Hialurônico/antagonistas & inibidores , Himecromona/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Inibidores da Angiogênese/farmacologia , Animais , Aorta/metabolismo , Aorta/patologia , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Endométrio/irrigação sanguínea , Endométrio/metabolismo , Endométrio/transplante , Feminino , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microvasos/patologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To evaluate the effect of anti-TNF-α in the treatment of endometrial implants in the peritoneum of rats. METHODS: Endometrial implants were surgically induced in 120 female Wistar-Albino rats. The animals were randomly divided into four groups. Group C (n = 36) received an intraperitoneal injection of 0.2 ml of saline. Group L (n = 41) received a subcutaneous injection of 1mg/kg of leuprolide. Group I5 (n = 20) received a subcutaneous injection of 5mg/kg of monoclonal anti-tumor necrosis factor (TNF) a (infliximab). Group I10 (n = 20) received a subcutaneous injection of 10mg/kg of infliximab. The rats were sacrificed after 21 days to assess the size of the implants and the expression of TNF. RESULTS: Treatment with leuprolide (group L) promoted an absolute reduction in the surface area of the implant when compared with group C (+14 mm vs. 0mm, p = 0.013) and group I10 (+14 mm vs. +5 Mm, p = 0.018). Likewise, a percentage reduction of surface area of the implant was observed comparing group L with group C (+33.3% vs. 0%, p = 0.005) and group I10 (+33.3% vs. +18.3%, p = 0.027). Treatment with infliximab was not able to decrease the surface area of the implants when compared with group C. The expression of TNF-α in groups L, I5 and I10 was lower than in group C (505.6 mm(2) vs. 660.5 mm(2) vs. 317.2 mm(2) vs. 2519.3 mm(2), respectively; p <0.001). CONCLUSION: The anti-TNF-α therapy reduced the expression of TNF-α in endometriotic implants, but did not reduce the surface area of the lesion.
Assuntos
Anticorpos Monoclonais/farmacologia , Endométrio/efeitos dos fármacos , Endométrio/transplante , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Animais , Feminino , Infliximab , Peritônio/cirurgia , Ratos , Ratos WistarRESUMO
OBJETIVO: Avaliar o efeito da terapia anti-TNF-α no tratamento de implantes endometriais no peritônio de ratas. MÉTODOS: Os implantes endometrióticos foram induzidos cirurgicamente em 120 ratas Wistar-Albino. Os animais foram aleatoriamente distribuídos em 4 grupos. O grupo C (n=36) recebeu uma injeção intraperitoneal de 0,2ml de solução salina. O grupo L (n=41) recebeu uma injeção subcutânea de 1mg/kg de leuprolide. O grupo I5 (n=20) recebeu uma injeção subcutânea de 5mg/kg de anticorpo monoclonal anti-fator de necrose tumoral (TNF) a (infliximab). O grupo I10 (n=20) recebeu uma injeção subcutânea de 10mg/kg de infliximab. As ratas foram sacrificadas após 21 dias para se avaliar o tamanho dos implantes e a expressão do TNF-α. RESULTADOS: O tratamento com leuprolide promoveu uma redução absoluta na área de superfície do implante comparado com o grupo C (+14mm vs. 0mm; p=0,013) e com o grupo I10 (+14mm vs. +5mm; p=0,018). Da mesma forma, uma redução percentual da area de superfície do implante foi observada comparando o grupo L com o grupo C (+33,3 por cento vs. 0 por cento; p=0,005) e com o grupo I10 (+33,3 por cento vs. +18,3 por cento; p=0,027). O tratamento com infliximab não foi capaz de diminuir a área de superfície do implante comparado com o grupo C. A expressão de TNF-α reduziu nos grupos L, I5 e I10 comparado com o grupo C (505,6µm² vs. 660,5µm² vs. 317,2µm² vs. 2519,3µm², respectivamente; p<0,001). CONCLUSÃO: A terapia anti-TNF-α reduziu a expressão de TNF-α nos implantes endometrióticos mas não reduziu a área de superfície da lesão.
OBJECTIVE: To evaluate the effect of anti-TNF-α in the treatment of endometrial implants in the peritoneum of rats. METHODS: Endometrial implants were surgically induced in 120 female Wistar-Albino rats. The animals were randomly divided into four groups. Group C (n = 36) received an intraperitoneal injection of 0.2 ml of saline. Group L (n = 41) received a subcutaneous injection of 1mg/kg of leuprolide. Group I5 (n = 20) received a subcutaneous injection of 5mg/kg of monoclonal anti-tumor necrosis factor (TNF) a (infliximab). Group I10 (n = 20) received a subcutaneous injection of 10mg/kg of infliximab. The rats were sacrificed after 21 days to assess the size of the implants and the expression of TNF. RESULTS: Treatment with leuprolide (group L) promoted an absolute reduction in the surface area of the implant when compared with group C (+14 mm vs. 0mm, p = 0.013) and group I10 (+14 mm vs. +5 Mm, p = 0.018). Likewise, a percentage reduction of surface area of the implant was observed comparing group L with group C (+33.3 percent vs. 0 percent, p = 0.005) and group I10 (+33.3 percent vs. +18.3 percent, p = 0.027). Treatment with infliximab was not able to decrease the surface area of the implants when compared with group C. The expression of TNF-α in groups L, I5 and I10 was lower than in group C (505.6 mm² vs. 660.5 mm² vs. 317.2 mm² vs. 2519.3 mm², respectively; p <0.001). CONCLUSION: The anti-TNF-α therapy reduced the expression of TNF-α in endometriotic implants, but did not reduce the surface area of the lesion.
Assuntos
Animais , Feminino , Ratos , Anticorpos Monoclonais/farmacologia , Endométrio/efeitos dos fármacos , Endométrio/transplante , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Peritônio/cirurgia , Ratos WistarRESUMO
OBJECTIVE: To characterize the pattern of cell proliferation and apoptosis of eutopic and ectopic endometrium in rabbits after endometrium implantation for the experimental induction of endometriosis. DESIGN: Animal experimental study. SETTING: Sector of experimental surgery. ANIMAL(S): Twenty-female New Zealand rabbits. INTERVENTION(S): All animals underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium, and fixation of tissue segment to the pelvic peritoneum. Two groups of 10 animals were sacrificed 4 and 8 weeks after endometriosis induction. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma determination. MAIN OUTCOME MEASURE(S): Cell proliferation and apoptosis were determined in the eutopic and ectopic endometrium, and the cell proliferation index (CPI) and apoptotic index (AI) were calculated as the number of labeled cells per 1,000 cells. The tissue homeostasis index was the CPI/AI ratio. Glands and stroma were analyzed separately. RESULT(S): The CPI for ectopic tissue was increased compared with eutopic tissue, but there was no difference in the ectopic lesions between 4 and 8 weeks of induction. Considering only the AI, eutopic and ectopic endometrium did not differ after 4 weeks, but differed significantly in glandular tissue after 8 weeks. The tissue homeostasis index revealed cell proliferation in these tissues, with a CPI/AI more than 1. CONCLUSION(S): Ectopic lesions seem to have a higher CPI than eutopic endometrium, with uncontrolled tissue growth occurring in induced endometriotic lesions.
Assuntos
Proliferação de Células , Endometriose/patologia , Endométrio/patologia , Animais , Apoptose , Modelos Animais de Doenças , Endométrio/transplante , Feminino , Homeostase , Coelhos , Células Estromais/patologia , Fatores de Tempo , Regulação para CimaRESUMO
PURPOSE: To evaluate macroscopically the growth degree of self-transplantation of endometriosis in rats. METHODS: Forty female rats, after a 7-day period for adpating and evaluating of the estrous cycle regularity, underwent tail abdominal midline laparotomy with 3-cm cuts. The average third of the left uterine horn was removed, 4mm x 4mm patches in liquid environment were made, and self-transplanted in the rat mesenterium with a single stitch, and the endometrial surface of the endometriotic implant facing the lumen of the peritoneal cavity. The rats were programmed to die after three weeks. The abdominal cavity displaying was held and self-transplants were identified and classified. RESULTS: The results achieved were: one case for degree 0 (2,5%), three cases for degree 1 (7,5%), eleven cases for degree II (27,5%) and twenty-five cases for degree III (62,5%). CONCLUSION: The experimental endometriosis development, through the self-transplantation technique, showed to be most common in degrees 3 and 2 of development.
Assuntos
Endometriose/patologia , Útero/transplante , Animais , Modelos Animais de Doenças , Endometriose/etiologia , Endométrio/citologia , Endométrio/transplante , Feminino , Ratos , Ratos Wistar , Transplante AutólogoRESUMO
PURPOSE: To evaluate macroscopically the growth degree of self-transplantation of endometriosis in rats. METHODS: Forty female rats, after a 7-day period for adpating and evaluating of the estrous cycle regularity, underwent tail abdominal midline laparotomy with 3-cm cuts. The average third of the left uterine horn was removed, 4mm x 4mm patches in liquid environment were made, and self-transplanted in the rat mesenterium with a single stitch, and the endometrial surface of the endometriotic implant facing the lumen of the peritoneal cavity. The rats were programmed to die after three weeks. The abdominal cavity displaying was held and self-transplants were identified and classified. RESULTS: The results achieved were: one case for degree 0 (2,5 percent), three cases for degree 1 (7,5 percent), eleven cases for degree II (27,5 percent) and twenty-five cases for degree III (62,5 percent). CONCLUSION: The experimental endometriosis development, through the self-transplantation technique, showed to be most common in degrees 3 and 2 of development.(AU)
OBJETIVO: Avaliar macroscopicamente o grau de crescimento de autotransplantes de endometriose em ratos. MÉTODOS: Quarenta ratos fêmeas, após período de sete dias para adaptação e avaliação da regularidade do ciclo estral, foram submetidas à laparotomia mediana abdominal caudal com incisões de três cm. Foi retirado o terço médio do corno uterino esquerdo, feito retalhos de quatro mm x quatro mm em meio líquido, sendo em seguida autotransplantado no mesentério da rata com ponto simples, tomando o cuidado de manter a superfície mucosa voltada para luz abdominal. Após o período de três semanas as mortes das ratas foram programadas. Realizou-se a exposição da cavidade abdominal com identificação e classificação dos autotransplantes. RESULTADOS: os resultados encontrados foram: grau 0 obteve um caso (2,5 por cento), o grau I foi observado em três casos (7,5 por cento), o grau II com onze casos (27,5 por cento) e o grau III foi visto em vinte cinco casos (62,5 por cento). CONCLUSÃO:Desenvolve-se a endometriose experimental pela técnica do autotransplante com a maioria dos casos em grau 3 e 2 de desenvolvimento.(AU)
Assuntos
Animais , Feminino , Ratos , Modelos Animais de Doenças , Endometriose/patologia , Útero/transplante , Endometriose/etiologia , Endométrio/citologia , Endométrio/transplante , Ratos Wistar , Transplante AutólogoRESUMO
PURPOSE: To evaluate macroscopically the growth degree of self-transplantation of endometriosis in rats. METHODS: Forty female rats, after a 7-day period for adpating and evaluating of the estrous cycle regularity, underwent tail abdominal midline laparotomy with 3-cm cuts. The average third of the left uterine horn was removed, 4mm x 4mm patches in liquid environment were made, and self-transplanted in the rat mesenterium with a single stitch, and the endometrial surface of the endometriotic implant facing the lumen of the peritoneal cavity. The rats were programmed to die after three weeks. The abdominal cavity displaying was held and self-transplants were identified and classified. RESULTS: The results achieved were: one case for degree 0 (2,5 percent), three cases for degree 1 (7,5 percent), eleven cases for degree II (27,5 percent) and twenty-five cases for degree III (62,5 percent). CONCLUSION: The experimental endometriosis development, through the self-transplantation technique, showed to be most common in degrees 3 and 2 of development.
OBJETIVO: Avaliar macroscopicamente o grau de crescimento de autotransplantes de endometriose em ratos. MÉTODOS: Quarenta ratos fêmeas, após período de sete dias para adaptação e avaliação da regularidade do ciclo estral, foram submetidas à laparotomia mediana abdominal caudal com incisões de três cm. Foi retirado o terço médio do corno uterino esquerdo, feito retalhos de quatro mm x quatro mm em meio líquido, sendo em seguida autotransplantado no mesentério da rata com ponto simples, tomando o cuidado de manter a superfície mucosa voltada para luz abdominal. Após o período de três semanas as mortes das ratas foram programadas. Realizou-se a exposição da cavidade abdominal com identificação e classificação dos autotransplantes. RESULTADOS: os resultados encontrados foram: grau 0 obteve um caso (2,5 por cento), o grau I foi observado em três casos (7,5 por cento), o grau II com onze casos (27,5 por cento) e o grau III foi visto em vinte cinco casos (62,5 por cento). CONCLUSÃO:Desenvolve-se a endometriose experimental pela técnica do autotransplante com a maioria dos casos em grau 3 e 2 de desenvolvimento.
Assuntos
Animais , Feminino , Ratos , Endometriose/patologia , Útero/transplante , Modelos Animais de Doenças , Endometriose/etiologia , Endométrio/citologia , Endométrio/transplante , Ratos Wistar , Transplante AutólogoRESUMO
Com o objetivo de estabelecer um modelo experimental de endometriose cirúrgica e de se estudar a açäo do danazol, valerianato de estradiol e ooforectomia, foram empregadas 48 ratas albinas submetidas a implante endometrial cirúrgico no peritônio lateral esquerdo, distribuídos em 04 grupos experimentais: controle, ooforectomia, danazol e valerianato de estradiol. Utilizou-se a citologia vaginal como parâmetro para indicaçäo do ato cirúrgico. O fragmento de endométrio foi obtido do corno direito e implantado no peritônio lateral esquerdo por meio de microcirurgia. O sacrifício dos animais foi realizado 30 dias após a cirurgia e realizado necroscópia de todos os órgäos. O material do implante foi estudado histopatologicamente. Durante o experimento analisou-se os seguintes parâmetros clínicos: citologia vaginal, glândulas mamárias e secreçäo vaginal. A análise dos resultados obtidos demonstrou exequibilidade do modelo experimental permitindo deduzir que o estímulo com o valerianato de estradiol mostre presença de tecido glandular com elevado índice de mitose, alteraçäo como hiperplasia do implante endometrial cirúrgico. Sob a influência do danazol os implantes apresentaram-se inviáveis, mostrando fibrose intensa
Assuntos
Ratos , Animais , Feminino , Danazol/metabolismo , Modelos Animais de Doenças , Endometriose/patologia , Estradiol/metabolismo , Endométrio/transplante , OvariectomiaRESUMO
O estudo foi realizado em 24 ratas albinas, divididas em 2 grupos experimentais, utilizando a técnica microcirúrgica para obtençäo do fragmento de endométrio e implantaçäo do fragmento endometrial no peritônio lateral esquerdo. Os implantes foram observados após 30 dias. Para a indicaçäo da operaçäo no 1§ grupo utilizou-se a induçäo prévia com a valerianata de estradiol e no 2§ grupo a citologia vaginal funcional na fase de estro terminal. Foi realizado estudo anatomopatológico do implante envolvendo a observaçäo da área, da vascularizaçäo, das aderências e retraçöes peritoniais, histologicamente foram analisadas as glândulas e estroma de padröes endometriais e análise das figuras de mitose no epitélio de revestimento glandular