Análise histomorfométrica no endométrio em modelo ectópico e eutópicos de endometriose em camundongas
Data
2021-08-26
Autores
Silva, Felipe Rosendo da 
Orientadores
Bertoncini, Clélia Rejane Antônio
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Objetivo: Realizar um modelo de endometriose em camundongos isogênicos Balb/c,
implantando tecido endometrial ectópico. Avaliar a histomorfometria dos tecidos endometriais
ectópico e eutópicos de camundongo controles e receptores. Método: Foram utilizados21
camundongos Balb/c fêmea entre 7 a 8 semanas, que foram divididos em 4 grupos, sendo que
primeiro grupo contém 3 camundongos doadores e o restante possuiu6 camundongos por
grupos. Grupo 1- Controle, são animais saudáveis não tiveram nenhum procedimento; Grupo
2-Sham, são animais que tiveram os mesmos procedimentos do animal modelo de
endometriose, porém não receberam o implante endometrial ectópico em seu lugar receberam
solução salina; Grupo 3- endometriose, os animais tiverem intervenções com implante
endometrial ectópico. Os camundongos do G1 e G3 foram tratados com 17 β estradiol diluídos
em óleo de rícino e tratados 1 vez por semana durante 4 semanas consecutivas. A administração
do 17 β estradiol foi utilizada para sincronizar a fase do ciclo estral. Após os tratamentos
aguardamos 2 semanas e no final realizamos a eutanásia dos camundongos doadores, retiramos
os cornos uterinos e o endométrio, para ser inoculado no grupo G3. Para tal procedimento
utilizamos cloridrato cetamina e xilazina como anestésico. Após este procedimento de indução
à endometriose, todos os animais foram eutanasiados para remoção dos cornos uterinos e o
endométrio que foram imersos em paraformaldeído a 4% para realização de processamento
histológico e coloração com H.E para obtenção das análises histomorfometricas e obtenção de
dados estatísticos. Resultado: Analisamos a área de superfície dos implantes e evidenciamos
importantes sítios de implantação de tecido endometrial na parede abdominal das
camundongas, com crescimento local de tecido, mostrando importantes lesões e aderências,
além de cistos escuros. Notamos que no grupo endometriose ocorreu um aumento no número
de vasos sanguíneos GIII e glandulares GIII (GIII ≥GI e GIII. P>0,001). Além do evidente
espessamento do epitelial endometrial GIII, quando comparados com o controle GI e Sham GII
(GIII ≥GI e GIII. P>0,001). Notamos também um aumento no número de eosinófilos(GIII (GIII
≥GI e GIII. P>0,001). Conclusão: O modelo empregado é de fácil execução e boa
reprodutibilidade, capaz de reproduzir as características morfológicas da endometriose, além
de possibilitar a obtenção de análises histomorfométricas do endométrio, mostrando que o
modelo reproduzido está relacionado a patogênese da endometriose.
Objective: To carry out a model of endometriosis in isogenic Balb/ mice, implanting ectopic endometrial tissue. To evaluate the histomorphometry of the ectopic and eutopic endometrial tissues of control and receptor mice. Method: 21 female Balb / mice between 7 and 8 weeks were used, which were divided into 4 groups, with the first group containing 3 donor mice and the remainder having 6 mice per group. Group 1- Control, healthy animals have not had any procedure; Group 2 -Sham, are animals that had the same procedures as the model animal of endometriosis, but did not receive the ectopic endometrial implant. In their place they received saline solution; Group 3 - endometriosis, the animals had interventions with an ectopic endometrial implant. The G1 and G3 mice were treated with 17 β estradiol diluted in castor oil and treated once a week for 4 consecutive weeks. The administration of 17 β estradiol was used to synchronize the phase of the estrous cycle. After the treatments, we waited 2 weeks and at the end we performed the euthanasia of the donor mice, removed the uterine horns and the endometrium, to be inoculated in the G3 group, for this procedure we used ketamine and xylazine hydrochloride as an anesthetic. After this endometriosis induction procedure, all animals were euthanized to remove the uterine horns and the endometrium that were immersed in 4% paraformaldehyde to perform histological processing and staining with H.E to obtain histomorphometric analyzes and obtain statistical data.Result: We analyzed the surface area of the implants and found important sites of implantation of endometrial tissue in the abdominal wall of the mice, with local tissue growth, showing important lesions and adhesions, in addition to dark cysts. We noticed that in the endometriosis group there was an increase in the number of blood vessels GIII and glandular GIII (GIII ≥GI and GIII. P> 0.001). In addition to the evident thickening of the GIII endometrial epithelial, when compared with the GI and Sham GII control (GIII ≥GI and GIII. P> 0.001). We also noticed an increase in the number of eosinophils (GIII (GIII ≥GI and GIII. P> 0.001). Conclusion: The model used is easy to perform and has good reproducibility, capable of reproducing the morphological characteristics of endometriosis in addition to enabling the achievement of histomorphometric analyzes of the endometrium, showing that the model reproduced is related to the pathogenesis of endometriosis.
Objective: To carry out a model of endometriosis in isogenic Balb/ mice, implanting ectopic endometrial tissue. To evaluate the histomorphometry of the ectopic and eutopic endometrial tissues of control and receptor mice. Method: 21 female Balb / mice between 7 and 8 weeks were used, which were divided into 4 groups, with the first group containing 3 donor mice and the remainder having 6 mice per group. Group 1- Control, healthy animals have not had any procedure; Group 2 -Sham, are animals that had the same procedures as the model animal of endometriosis, but did not receive the ectopic endometrial implant. In their place they received saline solution; Group 3 - endometriosis, the animals had interventions with an ectopic endometrial implant. The G1 and G3 mice were treated with 17 β estradiol diluted in castor oil and treated once a week for 4 consecutive weeks. The administration of 17 β estradiol was used to synchronize the phase of the estrous cycle. After the treatments, we waited 2 weeks and at the end we performed the euthanasia of the donor mice, removed the uterine horns and the endometrium, to be inoculated in the G3 group, for this procedure we used ketamine and xylazine hydrochloride as an anesthetic. After this endometriosis induction procedure, all animals were euthanized to remove the uterine horns and the endometrium that were immersed in 4% paraformaldehyde to perform histological processing and staining with H.E to obtain histomorphometric analyzes and obtain statistical data.Result: We analyzed the surface area of the implants and found important sites of implantation of endometrial tissue in the abdominal wall of the mice, with local tissue growth, showing important lesions and adhesions, in addition to dark cysts. We noticed that in the endometriosis group there was an increase in the number of blood vessels GIII and glandular GIII (GIII ≥GI and GIII. P> 0.001). In addition to the evident thickening of the GIII endometrial epithelial, when compared with the GI and Sham GII control (GIII ≥GI and GIII. P> 0.001). We also noticed an increase in the number of eosinophils (GIII (GIII ≥GI and GIII. P> 0.001). Conclusion: The model used is easy to perform and has good reproducibility, capable of reproducing the morphological characteristics of endometriosis in addition to enabling the achievement of histomorphometric analyzes of the endometrium, showing that the model reproduced is related to the pathogenesis of endometriosis.
Descrição
Citação
SILVA, F R. Análise histomorfométrica no endométrio em modelo ectópico e eutópicos de endometriose em camundongas. São Paulo, 2022. Dissertação (Mestrado em Ginecologia) - Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP). São Paulo, 2022.