两种宫腔粘连大鼠模型稳定性的比较

Exploration of the stability of two rat models of uterine adhesion

目的比较无水乙醇化学损伤法、刮宫与宫腔留置脂多糖双重损伤法建立SD大鼠宫腔粘连动物模型的造模效果,探索模型稳定性。方法110只SD雌鼠采用随机数表随机分为正常组、双重损伤组和无水乙醇组。正常组每组30只,余两组均为40只。无水乙醇组宫腔注射95%无水乙醇损伤子宫内膜,双重损伤组刮宫后宫腔留置脂多糖棉线2 d损伤子宫内膜,正常组行假手术。分别于术后1周、2周、4周、8周、12周采用随机数表每组各随机抽取3只动物收集子宫组织,行HE染色和Masson染色,对子宫内膜厚度、腺体数量和纤维化面积进行统计分析,并行免疫组织化学染色观察CD31在子宫内膜间质上的表达情况。结果①两种模型子宫内膜厚度在造模2周后,均小于正常组,且趋于稳定。无水乙醇组子宫内膜厚度较双重损伤组减少更明显(P均<0.05)。②两种模型子宫内膜腺体数量在造模1周后均明显小于正常组,无水乙醇组子宫内膜腺体数量较双重损伤组减少更明显(P均<0.05)。③两种模型子宫内膜间质纤维化面积在造模1周后均高于正常组(P均<0.05),但无水乙醇组纤维化面积在造模后8周开始减少。④两种模型子宫内膜间质CD31表达量在造模1周后均明显小于正常组(P均<0.05),无水乙醇组较双重损伤组减少更明显(P均<0.05)。⑤2周和12周模型鼠妊娠情况及胎鼠数目比较表明模型鼠生育能力均受到影响,且无水乙醇组模型鼠生育能力所受影响比双重损伤模型组明显。结论两种方法均能诱导宫腔粘连;无水乙醇化学损伤法操作较简单、经济,但纤维化稳定性欠佳,其子宫内膜厚度、腺体数量及血管改变符合薄型子宫内膜病理变化,更适用于薄型子宫内膜疾病的相关研究;双重损伤模型贴近临床损伤方法,且模型更稳定,可以满足宫腔粘连相关研究需求。

Objective To compare the modeling effects of anhydrous ethanol chemical injury method and uterine curettage combine uterine lipopolysaccharide indwelling double injury method,and to explore the stability of the models.Methods Totally 110 SD female rats were randomly divided into normal group(n=30),dual injury group(n=40)and alcohol group(n=40)by using a random number table.In the alcohol group,95%absolute ethanol was injected into the uterine cavity to damage the endometrium,and in the dual injury group,lipopolysaccharide cotton thread was retained in the uterine cavity for 2 d after uterine curettage,and sham operation was performed in the normal group.After retrieval of the uteri on the 1st,2nd,4th,8th and 12th weeks post-procedure,they were embedded in paraffin,serially sectioned at 5µm thickness and routinely stained with hematoxylin and eosin and masson stains.The sections obtained from the samples collected from the above were also used to detect the expression of CD31(a vascular marker)on the uterine tissues by immunohistochemistry staining.Results 1)Two weeks after operation,the endometrial thickness of the two models were smaller than that of the normal group,and tended to be stable.The thickness of endometrium in alcohol group decreased more significantly than that in dual injury group(all P<0.05).2)One week post-procedure,the number of endometrial glands in the two models was significantly less than that in the normal group,and it was more obvious in the alcohol group than that in the dual injury group(all P<0.05).3)The area of endometrial interstitial fibrosis in the model groups were both higher than that in the normal group from one week after operation(all P<0.05),but the fibrosis area in the alcohol group decreased from the 8th week.4)One week post-procedure,the expression of CD31 in the endometrial stroma of the two models were significantly lower than that in the normal group(P<0.05),and the decrease in the alcohol group was more obvious than that in the dual injury group(all P<0.05).5)The comparison of the pregnancy ability and the number of fetal mice between 2-week and 12-week model mice showed that the fertility of the model mice was affected,and the fertility of the alcohol group was more affected than that of the dual injury model group.Conclusion The two methods can both induce uterine adhesion.Alcohol injury method is simple and economical,but the stability of fibrosis is poor,and the thickness of endometrium,the number of glands and the changes of blood vessels accord with the pathological changes of thin endometrium,which is more suitable for the study of thin endometrial diseases.The dual injury model is close to the clinical injury method,and the model is more stable,which can meet the needs of uterine adhesion related research.