动物造模 z-bio 2022-10-19 312

　　Objective To establish an animal model of intrauterine adhesions according to the characteristics of clinical injury by using mechanical injury combined with lipopolysaccharide perfusion in a minimally invasive manner, so as to provide support for the in-depth study of the pathogenesis, pathological changes and exploration of clinical treatment plans of IUA.

　　Methods Twenty female ICR mice were randomly divided into two groups (n=10). After phloroglucinol relaxed the cervix, the model group animals were given intrauterine mechanical injury+lipopolysaccharide (LPS) perfusion (0.5mg/kg) through the cervix, and the sham operation group animals were only given equal volume of normal saline perfusion. Uterine specimens were collected at 7, 14, and 21 days after operation. Routine HE and Masson staining were used to detect endometrial morphology and fibrosis; Immunofluorescence was used to detect the distribution of macrophages (F4/80) in the endometrium, and qPCR and Western Blot were used to detect the inflammatory factor (II-1) in the endometrium of 14 day old mice β、 TNF- α、 IL-4, IL-10 and IL-6) and endometrial receptivity related markers (Integrin B3 and LIF). Subsequently, 20 mice were randomly divided into mechanical injury via cervix+LPS perfusion group (n=10) and sham operation group (n=10). After 2 weeks of modeling, the cage was closed and the number of pregnant rats was recorded; The female rats were killed about 18 days after pregnancy, and the number of embryos was recorded.

　　Results After mechanical injury and LPS perfusion, the inflammatory reaction and fibrosis degree in mouse uterus were significantly increased. The thickness of endometrium in the model group was significantly lower than that in the sham operation group, with obvious necrosis of endometrial cells, loss of glands, incomplete structure of luminal epithelium and glandular epithelial cells, and severe tissue fibrosis. Subsequently, it was found that the expression of macrophage marker F4/80 in mouse endometrium of model group was significantly increased (P<0.05), and pro-inflammatory factor II-1 in mouse endometrium of model group β、 TNF- α、 The expression of IL-4 and IL-10 decreased significantly (P<0.05). Further study found that endometrial receptivity markers LF and Integrin β 3 The expression level decreased significantly (P<0.05), and the number of pregnant rats and fetal rats decreased significantly (P<0.05).

　　Conclusion This study successfully established a mouse IUA model construction method by minimally invasive cervical mechanical injury+lipopolysaccharide perfusion, which is in line with the characteristics of clinical IUA injury, and can provide a stable animal model for the follow-up study on the pathogenesis and treatment of IUA.