标记滞留细胞技术检测异位子宫内膜干细胞的研究  

The study of identification of label-retaining cells in mouse model of endometriosis

作　　者：王宇全[1] 尹利荣[1] 郭蕊萌[1] 

机构地区：[1]天津医科大学第二医院妇科,天津300211

出　　处：《现代妇产科进展》2013年第2期106-110,114,共6页Progress in Obstetrics and Gynecology

基　　金：天津医科大学资助课题(No:2010ky49)

摘　　要：目的:通过标记滞留细胞(LRCs)技术检测证实异位子宫内膜干细胞的存在及其在病灶内的分布情况;初步探讨抗雌激素治疗对异位子宫内膜干细胞的影响,寻找内异症高复发率的原因。方法:将64只裸鼠分成3组:对照组16只,建立内异症裸鼠模型前24h开始腹腔注射NS 50μl/(只.次),2次/d,共5d;未去势组及去势组各24只,建模前24h开始腹腔注射25μg/μl 5-溴脱氧尿苷(BrdU),2μl/g,2次/d,共5d,去势组于末次注射后1w去势。造模后裸鼠均雌激素刺激5d。3组裸鼠均于于末次注射后4h,1w,3w,5w,6w,7w,8w,9w进行探查,对照组每次处死2只裸鼠,其余2组每次处死3只裸鼠。免疫组化法检测每个时间点的BrdU标记率。结果:BrdU标记未对造模结果产生不良影响。对照组各时间点均未见BrdU标记阳性细胞。未去势组的腺上皮和间质的BrdU初始最大标记率分别为(69.89±0.78)%和(73.27±1.06)%,第6w和8w时分别约有(2.07±0.19)%和(4.63±0.72)%仍保留标记,分别为腺上皮和间质LRCs,分别于第7w和9w时衰减至0。腺上皮和间质LRCs多位于异位内膜与所种植的裸鼠组织的交界处,约50%的间质LRCs临近血管和腺体。去势治疗可显著减小异位病灶的体积,但对LRCs的数量无明显影响。去势组与未去势组的腺上皮、间质标记率在各个时间点均无统计学差异(P>0.05)。结论:LRCs技术可以成功标记异位子宫内膜干细胞,异位病灶中存在腺上皮和间质两种LRCs,抗雌激素治疗并不能清除LRCs。干细胞的持续存在可能在内异症的复发机制中扮演了重要角色。Objective：To identify adult stem cells and their locations in ectopic endo- metrium by detecting label-retaining cells （LRCs） using the 5-bromo-2-deoxyuridine （BrdU） labeling method. To explore the cause of high recurrence rate in endometriosis by oberving the effect of anti-estrogen therapy on LRCs in ectopic endometrium. Method：64 female nude mice were divided into 3 groups. In the control group（ n= 16） ,501xl NS was given twice daily for 5 consecutive days by intraperitoneal injection,24h before the endometriosis model established. In the no castration and castration groups （n = 24, each）, BrdU （25 Ixg/Ixl, 501xg/g of body weight per time） was given twice daily for 5 consecutive days by intraperitoneal injection,24h before the endometriosis model established. The mice in castration group were castrated at the time of 1 w after the last injection. Chase periods were 4h, 1 w,3w, 5w,6w,7w, 8w, 9w when the last in- jection was finished. At each time of point 3 mice were sacrificed in no castration and castration groups and 2 mice were sacrifeced in control group to get the BrdU labeling rate by immunohis- tochemistry. Result： No adverse effect of BrdU labeling was detected on endometriosis model mice. The initial BrdU labeling rates were （69.89±0.78） % and （73.27±1.06） % for epithe- lial and stromal respectively in no castraion group. After a 6-week and 8-week chase, （2.07±0. 19） % epithelial cells and （4.63±0.72）% stromal cells retained BrdU labeling respectively. Epithelial and stromal LRCs were considered to be adult stem cells. Both epithelial and stromal LRCs were mainly located in the boundary of the lesions. About 50% stromal LRCs were adja- cent to glands and blood vessels. Castration could remarkably decrease the lesion＇s volum, but had no effect on the numbers of LRCs. No difference was found between the no castration group and castration group of the BrdU labeling rates in glands and stroma at each time of chase point （P〉0.05）. Conclusion：LRCs in e

关 键 词：干细胞 子宫内膜 小鼠 近交系 染色与标记 免疫组织化学 溴脱氧尿苷 

分 类 号：R711.71[医药卫生—妇产科学]