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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 detect the presence of ectopic endometrial stem cells and its distribution in the lesion by detecting labeled residual cells (LRCs) technique. To investigate the effect of anti-estrogen treatment on ectopic endometrial stem cells and to find out the high endometriosis Reasons for the recurrence rate. Methods: Sixty-four nude mice were divided into three groups: control group (n = 16), intraperitoneal injection of NS 50μl / (only) twice a day for 5 days before establishment of endometriosis nude mice model; The rats in castrated group were injected intraperitoneally with 25 μg / μl BrdU, 2 μl / g twice a day for 5 days before castration, and castrated at 1 w after the last injection. Nude mice were stimulated with estrogen 5 days after modeling. Three groups of nude mice were probed at 4h, 1w, 3w, 5w, 6w, 7w, 8w and 9w after the last injection. Two nude mice were sacrificed in the control group and three nude mice were sacrificed at the same time. Immunohistochemistry was used to detect the BrdU labeling rate at each time point. RESULTS: The BrdU labeling did not adversely affect the modeling results. BrdU positive cells were not found in control group at all time points. The initial maximal labeling rates of BrdU in glandular epithelium and stroma were (69.89 ± 0.78)% and (73.27 ± 1.06)% in un-castrated group and (2.07 ± 0.19)% and (4.63 ± 0.72)% of the remaining markers, respectively glandular epithelial and interstitial LRCs, decay to 0 at 7w and 9w respectively. Glandular epithelial and interstitial LRCs are mostly located at the junction of the ectopic endometrium and the implanted nude mice tissues, and approximately 50% of interstitial LRCs are adjacent to blood vessels and glands. Castration treatment significantly reduced the volume of ectopic lesions, but had no significant effect on the number of LRCs. There was no significant difference in the glandular epithelial and mesenchymal marker rates between the castrated group and the non-castrated group at all time points (P> 0.05). CONCLUSION: LRCs can successfully label ectopic endometrial stem cells. There are two kinds of LRCs in gland epithelium and stroma in ectopic lesions. Anti-estrogen therapy can not clear LRCs. The persistence of stem cells may play an important role in the mechanism of recurrence of endometriosis.