Establishment of a mouse intrauterine adhesion model by transcervical mechanical injury combined with lipopolysaccharide perfusion
Received:October 25, 2021  
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DOI:10. 3969 / j.issn.1005-4847. 2022. 03. 013
KeyWord:intrauterine adhesion; mechanical injury; lipopolysaccharide; animal model
常品 1. 新乡医学院第三附属医院,河南 新乡 ;2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡
张胜辉 1. 新乡医学院第三附属医院,河南 新乡 ;2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡
陈童彤 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 ; 3. 新乡医学院生命科学技术学院,河南 新乡
孙钰椋 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 ;4. 新乡医学院医学工程学院,河南 新乡
张花 新乡医学院第三附属医院,河南 新乡
潘莹 新乡医学院第三附属医院,河南 新乡
刘彦礼 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 ; 3. 新乡医学院生命科学技术学院,河南 新乡
林俊堂 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 ;4. 新乡医学院医学工程学院,河南 新乡
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       Objective To establish a stable animal model of intrauterine adhesion ( IUA) using a minimally invasive method that recapitulates the clinicopathologic characteristics of IUA. Methods Overall, 20 female ICR mice were randomly divided into two groups (n= 10). After the cervix was relaxed with phloroglucinol, the uterine horn of mice in the model group was subjected to mechanical injury and lipopolysaccharide perfusion (0. 5 mg / kg) through the cervix to induce endometrial injury, while mice in the sham operation group only received the same volume of normal saline perfusion. Uteri of the mice were collected at 7, 14 and 21 days after the procedure. HE and Masson staining were used to assess uterine morphology and fibrosis. Immunofluorescence was performed to evaluate macrophage infiltration into the endometrium. qPCR and Western Blot were used to detect the expression of inflammatory factors ( IL-1β, TNF-α, IL-4, IL-10 and IL-6) and endometrial receptivity-related markers ( integrin β3 and LIF) in mice at 14 days after modeling. Furthermore, mouse fertility was evaluated in model and sham operation groups. Results Compared with the sham operation group, endometria in the model group were significantly thinner and exhibited severe necrosis, glandular loss, and incomplete luminal epithelial and glandular epithelial cell structures. Subsequently, severe tissue fibrosis was observed in endometria in the model group. Additionally, increased macrophage infiltration and upregulated expression of proinflammatory factors demonstrated an activated inflammatory response in endometria of the model group. Moreover, endometrial receptivity was significantly decreased after mechanical injury combined with lipopolysaccharide perfusion. Finally, significant decreases in pregnancy and the number of fetuses were observed in the model group. Conclusions This study demonstrated successful establishment of an IUA mouse model through a minimally invasive transcervical mechanical injury combined with lipopolysaccharide perfusion, which was consistent with the clinical characteristics of severe IUA. This model might support in-depth study of IUA pathogenesis and promote the development of IUA therapies.
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