异种移植裸鼠子宫内膜异位症纤维化模型的构建

Establishment of endometriosis fibrosis nude mice model using xenotransplantation of human endometrium

目的探讨采用子宫内膜异位症患者的子宫内膜组织建立纤维化裸鼠模型的可行性,比较不同注射方法对建模成功率的影响。方法24只BALB/c裸鼠分为两组,每组12只建立子宫内膜异位症纤维化模型。收集子宫内膜异位症患者的在位内膜组织,使用皮下注射及腹腔注射方法将子宫内膜组织碎片种植进入裸鼠体内,种植后第14天处死裸鼠,观察异位组织与周围粘连情况,HE染色验证建模成功,Masson染色可见蓝色胶原纤维,天狼星红染色可见红色胶原纤维,免疫组化明确纤维化相关蛋白Collagen1高表达,普鲁士蓝染色证实组织中存在铁离子沉积。结果两种注射方法均可成功构建子宫内膜异位症裸鼠纤维化模型,12只裸鼠通过皮下注射方法建模,10只成功;12只裸鼠通过腹腔注射方法建模,2只成功。异位组织结节质硬,与周围粘连,活动度差,镜下可见结节内存在子宫内膜样腺体增生、胶原纤维沉积。证实子宫内膜异位症纤维化模型建立成功后,对建模成功裸鼠的异位组织进行铁离子检测,显示病灶中存在铁离子沉积。结论与腹腔注射建模方法相比,皮下注射方法构建异体裸鼠子宫内膜异位症纤维化模型成功率更高,建模成功的裸鼠异位组织中存在铁离子沉积现象,为进一步探索子宫内膜异位症纤维化研究提供了基础。

Objective To explore the feasibility of animal model of endometriosis fibrosis in nude mice using endometrium of endometriosis patients,and compare the influence of different injection methods on the effect of transplantation.Methods 24 BALB/c nude mice were divided into two groups with 12 mice in each group to establish the fibrosis mice model of endometriosis.The eutopic endometrial tissue of patients with endometriosis was collected,and the fragments of human endometrial tissue were grafted subcutaneously and intraperitoneally into the BALB/c nude mice respectively.The mice were sacrificed on the 14th day after implantation to verify the successful establishment of mice model by HE staining.The adhesion was observed,the collagen fiber was stained in blue by Masson's trichrome staining,red-stained collagen fiber was deceted by sirius red staining,and immunohistochemistry staining was used to evaluate the expression of Collagen 1,which was considered as fibrosis-related protein.The iron was deceted by prussian blue staining.Results The fibrosis model of endometriosis in nude mice was successfully established by both subcutaneous and intraperitoneal injection of human endometrium fragments.There are ten of twelve and two of twelve mice were successfully established by subcutaneous and intraperitoneally injection,respectively.The ectopic tissue was hard,adherent to the surrounding area,and had poor mobility.Endometrium-like glands,collagen deposition and iron deposition were found in ectopic tissues,which confirming the success establishment of mice model and the existence of iron in ectopic tissue.Conclusions The subcutaneous injection method has a higher success rate in the establishment of endometriosis fibrosis in nude mice compared with intraperitoneal injection,this study provides a model application for further exploration of endometriosis fibrosis and confirmed the iron deposition in ectopic tissue.