Histomorphometric analysis of the endometrium in an ectopic model of endometriosis in mice

Abstract

Objective: Evaluate histomorphometry of ectopic and eutopic endometrial tissues in receptor mice. Method: Eighteen female Balb/c were divided into 3 groups, 6 animals each: GI Control, no procedure; GII – Sham, animals that had the same procedures as GIII without receiving the ectopic endometrial implant. Instead, they received saline solution; GIII – endometriosis model, animals had surgical intervention with an ectopic endometrial implant. GI and GIII mice were treated with 17β-estradiol, 100 µg/kg each. All animals were euthanized to collect uterine horns, which were fixed in 4% paraformaldehyde, embedded in paraffin, stained with Hematoxilin and Eosin and submitted to histomorphometric analyzes. Data underwent one-way ANOVA followed by Tukey’s test. Results: Local tissue growth, showing important lesions and adhesions, as well as dark cysts were noticed. In GIII group, there was an increase in number of blood vessels and glands (GIII ≥ GI and GIII p > .001). Thickening of the GIII endometrial epithelial was also evident (GIII ≥ GI and GIII. p > .001). We also noticed an increase in the number of eosinophils (GIII (GIII ≥ GI and GIII. p > .001). Conclusion: Easy to perform model, capable of reproducing morphological endometriosis characteristics. From our findings, there was an increase of endometrial thickness as well as an increase in the eosinophils population.

摘要

目的

评价受体小鼠异位和在位子宫内膜的组织形态学。

方法

18只雌性Balb/c小鼠分为3组, 每组6只：GI对照组, 不做任何处理；GII-假手术组, 采用与GIII组相同的程序, 但未植入异位子宫内膜, 而是用生理盐水取而代之；GIII-子宫内膜异位症模型, 手术植入异位子宫内膜进行干预。GI和GIII组小鼠按100µg/kg给予17β-雌二醇。所有小鼠安乐死后采集子宫角, 用4%多聚甲醛固定, 石蜡包埋, 用苏木精和伊红染色, 进行组织形态学分析。数据用图基检验进行单因素方差分析。

结果

观察局部组织的生长, 有无明显的损伤和粘连, 有无黑色囊肿。GIII组血管和腺体数量增加（GIII≥GI和GII, p>0.001）。GIII组子宫内膜上皮增厚明显（GIII≥GI和GII, p>0.001)。我们还注意到嗜酸性粒细胞数量增加（GIII≥GI和GII, p>0.001）。

结论

模型制作简便, 能重现子宫内膜异位症形态学特征。从我们的发现来看, 嗜酸性粒细胞数量增加时, 伴有子宫内膜厚度增加。

Keywords:

• Mouse
• histomorphometry
• female infertility

关键词:

• 小鼠
• 组织形态学
• 女性不孕

Acknowledgements

We would like to thank Mauricio Rosa Trotta, from Center for the Development of Animal Models for Biology and Medicine (CEDEME – UNIFESP) for aiding us with animal care. We would like to thank National Council for Scientific and Technological Development (CNPq) for the grant to Felipe Rosendo da Silva.

Authors’ contributions

Felipe Rosendo da Silva contributed to brainstorming, manuscript writer, data gathering, main researcher. Daniel Soares Thimoteo contributed to data gathering. Adriana Ferraz Carbonel contributed to brainstorming, scientific advisor, manuscript writer, data gathering. Luiz Fernando Portugal Fuchs contributed to manuscript revisor. Gisela Rodrigues da Silva Sasso contributed to data analyzing, manuscript revisor. Ricardo dos Santos Simões contributed to data evaluating, manuscript revisor. Adriana Invitti contributed to review & editing. Renata Ramos Vieira contributed to data analyzing. Luiz Philipe de Souza Ferreira contributed to English review. Rafael André da Silva contributed to English review. Patricia Daniele Azevedo Lima contributed to data acquiring and analyzing. José Maria Soares Júnior contributed to scientific advisor. Manuel de Jesus Simões contributed to scientific advisor, manuscript revisor. Clélia Rejane Antonio Bertoncini contributed to brainstorming, manuscript writer, data evaluating.

Disclosure statement

No potential conflict of interest was reported by the authors.