建立裸鼠皮下荧光子宫内膜异位症模型

Establishing a nude mouse model of green fluorescence endometriosis

背景:将绿色荧光蛋白标记的裸鼠子宫内膜注射于裸鼠皮下,构建子宫内膜异位症动物模型,操作简便,同时可在一定时间内体外动态观察异位病灶的生长,有利于子宫内膜异位症的深入研究。目的:探讨构建裸鼠皮下子宫内膜异位症荧光模型的方法。方法:取裸鼠子宫内膜,采用增强型绿色荧光蛋白腺病毒转染后,注射入 6 只裸鼠皮下(转染荧光组),将未接受增强型绿色荧光蛋白腺病毒转染的子宫内膜组织块注射到另 6 只裸鼠腹部皮下(对照组)。于注射后第 5,10,15,20,25,30 天在体视荧光显微镜下观察皮下绿色荧光强度,记录荧光标记病灶的持续时间,同时行组织学检测两组异位病灶的情况。结果与结论:成功构建 5 个内膜异位症活体荧光裸鼠模型及 5 个内膜异位症活体非荧光裸鼠模型,平均每只裸鼠形成二三个病灶。利用体视荧光显微镜可通过活体动物的皮肤直接观察到体内病灶生长和发展,荧光面积和强度随时间延长逐渐减弱,可持续观察 4 周左右。对照组未检测到绿色荧光病灶。获取病灶组织经苏木精-伊红染色,可观察到典型的子宫内膜腺体和基质结构。结果可见这种改良的裸鼠荧光模型可以动态无损伤量化观察病灶生长和发展情况,有助于内膜异位症新药的开发及其发病机制的研究。

BACKGROUND： It is simple to fabricate an animal model of endometriosis by subcutaneous injection of green fluorescence labeled endometrium of nude mice. Meanwhile, in vitro ectopic lesions can be observed dynamically within a certain period, which contributes to the depth study of endometriosis. OBJECTIVE： To explore the method to establish an animal model of subcutaneous endometriosis in nude mice. METHODS： Enhanced green fluorescent protein adenovirus transfected endometrium of nude mice was injected subcutaneously into six nude mice （fluorescent group） and non-transfected endometrium was injected into another six nude mice （control group）. Green fluorescence intensity, duration of fluorescencelabeled lesions, and histological changes of ectopic lesions in vivo were detected under stereologica fluorescence microscope at 5, 10, 15, 20, 25, 30 days after injection. RESULTS AND CONCLUSION： Five living fluorescence mouse models of endometriosis were induced successfully in the fluorescent group and another five living non-fluorescent nude mouse models of endometriosis were also prepared in the control group. There were 2 3 lesions per nude mouse. Development of ectopic lesions in the mice could be monitored under the stereological fluorescence microscope. The fluorescent area and intensity were decreased with time, which lasted for about 4 weeks. Fluorescence labeled lesions were invisible in the control group. Hematoxylin-eosin staining showed the typical endometrial glands and stroma structure. These findings indicate that this modified nude mouse model can enable the dynamic and quantitative observation of lesion growth and development without injury and contribute to the development of new drugs for endometriosis and the underlying mechanism of endometriosis.