动物造模,宫腔黏连模型 z-bio 2022-09-05 369

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

　　Methods Twenty female ICR mice were randomly divided into 2 groups (n=10). After phloroglucinol relaxes the cervix, the animals in the model group were given intrauterine mechanical injury + lipopolysaccharide (LPS) perfusion (0.5 mg/kg) through the cervix. kg), and the animals in the sham-operated group received only an equal volume of normal saline perfusion. Uterine specimens were collected at 7, 14, and 21 days after operation, and 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, qPCR and Western Blot detection of inflammatory factors (II-1β, TNF-α, IL-4, IL-10 and IL-6) and endometrial receptivity-related markers (Integrin B3 and LIF) in the endometrium of model mice at 14 days expression changes. Then, 20 mice were randomly divided into transcervical mechanical injury + lipopolysaccharide perfusion group (n=10), modeling group and sham operation group (n=10). After 2 weeks of modeling, they were put into cages, and the number of pregnant female mice was recorded; The female mice were sacrificed at about 18 days of gestation, and the number of embryos was recorded.

　　Results The inflammatory response and fibrosis in the uterus of mice were significantly increased after mechanical injury + lipopolysaccharide perfusion. The endometrial thickness of the mice in the model group was significantly lower than that in the sham-operated group, with obvious endometrial cell necrosis, loss of glands, incomplete luminal and glandular epithelial cell structures, and severe tissue fibrosis. Subsequently, it was found that the expression of macrophage marker F4/80 in the endometrium of mice in the model group was significantly increased (P<0.05), and the pro-inflammatory factors II-1β, TNF-α, The expression of L-6 and L-6 were significantly increased (P<0.05), while the expressions of anti-inflammatory factors IL-4 and IL-10 were significantly decreased (P<0.05). Further research found that the expression levels of endometrial receptivity markers LF and Integrin β3 were significantly decreased (P<0.05), and the fertility test results showed that the number of pregnant mothers and fetuses were significantly decreased (P<0.05).

　　Conclusion In this study, a minimally invasive method of cervical mechanical injury + lipopolysaccharide perfusion was used to successfully establish a mouse IUA model construction method, which is in line with the clinical characteristics of IUA injury, and can provide a stable animal model for subsequent research on the pathogenesis and treatment of IUA.