用模拟经期内膜组织建立子宫内膜异位症小鼠模型及其生殖能力的研究

目的建立子宫内膜异位症(EM)模型小鼠,观察EM模型小鼠的生殖能力,探讨影响其生殖能力的可能原因。方法将51只8周龄雌性C57BL/6小鼠作为受体,并随机分为模型组(n=21)、假手术组(n=15)和空白组(n=15)。用模拟小鼠经期内膜组织腹腔注射的方法建立EM模型;将同等剂量的供体腹腔脂肪组织碎片注入假手术组小鼠的腹腔中;空白组不进行腹腔注射。观察3组小鼠的动情周期,于动情间期采血。将3组小鼠合笼至出现阴栓,统计各组小鼠的胚胎种植数、妊娠率、吸收胎率等。运用酶联免疫吸附法(ELISA法)检测各组小鼠血清FSH、LH、糖类抗原125(CA125)浓度及腹腔液中白介素-2(IL-2)水平,比较各组间的水平差异。结果成功建立EM小鼠模型,模型组小鼠动情周期规律。模型组小鼠的妊娠率显著低于假手术组、空白组(P0.05);3组小鼠的吸收胎率、畸胎率、胚胎重量、顶臀径等比较无显著性差异(P0.05)。模型组不孕鼠的血清FSH浓度、腹腔液IL-2浓度显著高于妊娠实验前的模型组、假手术组和空白组(P0.01);血清LH、CA125浓度在各组间无显著性差异(P0.05)。结论用模拟经期内膜组织腹腔注射的方法可以成功建立EM模型,模型组不孕鼠的卵巢功能、腹腔免疫微环境受到了影响,可能是造成生殖能力下降的重要原因。

Establishment of mouse endometriosis model by injecting simulated menstrual endometrial tissue and its reproductive ability

Objective: To establish mouse model of endometriosis (EM) to observe its fertility,and explore the possible factors affecting the reproductive ability. Methods: Fifty one female mice aged 8 weeks were used as recipient mice and randomly divided into model group (n=21),sham operation group (n=15) and blank group (n=15). The model mice were established by intraperitoneal injection of simulating menstrual endometrial tissue of donor mice. The mice of sham group were established by injecting fragments of abdominal adipose tissue from donor mice into peritoneal cavity. The mice of blank group were not injected intraperitoneally. The three groups of mice were observed in estrus cycle,and the blood samples were collected at diestrus. The mice of three groups were mated with male mice to appear vaginal plug. The number of embryos,pregnancy rate,absorptive fetus rate and so on were counted in each group of mice. The serum levels of FSH,LH,carbohydrate antigen 125 (CA125) and interleukin-2 (IL-2) in the peritoneal fluid were measured by enzyme-linked immunosorbent assay (ELISA) and compared among the groups. Results: The mouse model of EM was successfully established. The estrous cycle of model group was normal. The pregnancy rate of model group is significantly lower than the sham group and blank group (P<0.05). However there were no significant differences in absorptive fetus rate,teratogenic rate,pup weight,and crown-rump length among the three groups (P>0.05). The serum FSH levels and IL-2 concentration of infertile mice in model group were significantly higher than the three groups before mating (P<0.01). There were no statistical differences in the levels of serum LH and CA125 among the groups (P>0.05). Conclusions: The mouse endometriosis model is successfully established. In the model mice,the ovarian function and the peritoneal immune microenvironment were affected in infertile mice,which may be the important reason for the decrease of the reproductive ability.