异位子宫内膜干细胞的检测及其标志物研究

The Study of Identification of Ectopic Endometrial Stem Cells and Its Markers in Mouse Model of Endometriosis

目的了解异位子宫内膜干细胞在病灶中的数量、分布及定位情况,并筛选其可能的标志物表达。方法在子宫内膜异位症裸鼠模型建造前24h开始进行5-溴脱氧尿苷(BrdU)脉冲标记,共5d。在标记结束后4h、1周、3周、5周、6周、7周、8周及9周,通过免疫组化检测异位病灶中BrdU的表达情况,确定标记滞留细胞(The label-retaining cells,LRCs)。通过免疫组化双染检测LRCs表面CD34和CD146的表达情况。结果腺上皮和间质的BrdU初始最大标记率分别为(69.89±0.78)%和(73.27±1.06)%,于第6周和8周时分别有约(2.07±0.19)%和(4.63±0.72)%的腺上皮和间质细胞仍保留标记,即为上皮和间质标记滞留细胞,多位于异位内膜与所种植的裸鼠组织的交界处。LRCs并不表达CD34,少数间质LRCs表达CD146。结论异位子宫内膜中有上皮和间质两种干细胞,主要定位于异位内膜与所种植的组织的交界处,CD34不是异位内膜干细胞的标志物,CD146表达于异位内膜干细胞,但并非其特异性标志物。

Objective To identify and locate adult stem cells in ectopic endometrium and to find their markers in en- dometriosis. Methods The female nude mice were pulse labeled using 5-bromo-2-deoxyuridine （BrdU） for 5 consecutive days, 24 h prior to the endometriosis rat model was established. The label-retaining cells （LRCs） were identified by detecting BrdU labeling rate using immunohistochemistry on 4 h, 1-week, 3-week, 5-week, 6-week,7-week, 8-week and 9-week after injection. The expression levels of CD34 and CD146 on LRCs were detected by double staining of immunohistochemistry.Resuits The maximum BrdU labeling rates were （69.89±0.78）% and （73.27±1.06）% for epithelial and stromal cells respective- ly. 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, which were considered to be epithelial and stromal LRCs. Both the epithelial LRCs and the stromal LRCs were mainly located in the boundary of the lesions. LRCs did not co-express CD34. CD146 co-expressed on a few stromal LRCs. Conclusion Two kinds of stem cells were identified successfully in ectopic endometrium by label-retaining cell approach, which were epithelial and stromal LRCs, both located in the boundary of the lesions. It seems that CD34 is not the marker for ectopic endometrial stem cells. C D 146 maybe the candidate marker for ectopic endometrium but without specificity.