应用小鼠月经模型对子宫内膜血管周期性变化的形态学研究

Study of the cyclic changes of endometrial vasculatures with murine menstruation model

目的应用小鼠月经模型,研究子宫内膜血管形态学和血管内皮细胞在子宫组织中所占体积百分比(volume fraction)的月经周期变化. 方法成年雌性去势C57BL/6小鼠34只,经性激素处理后,实验组小鼠宫腔内注射0.2 ml消毒花生油诱导子宫内膜蜕膜化反应;对照组无宫腔油剂注射.取各组子宫组织,行常规HE染色,并用CD31单克隆抗体作为血管内皮细胞的标记分子进行免疫组织化学染色,计算机辅助图像分析软件CAST2,计算分析血管内皮细胞在子宫组织中所占体积百分比. 结果实验组宫腔注射油剂后30～35 h观察到子宫内膜脱落,内膜血管呈双层结构,近肌层的血管组织结构保持正常,近内膜的血管结构紊乱;对照组子宫内膜始终完整,但全层血管结构紊乱,未见分层现象.注射油剂后56～70 h实验组子宫内膜上皮基本修复,但血管稀疏,结构紊乱;对照组血管均完全修复.血管内皮细胞在子宫组织所占体积百分比在注射油剂56～70 h后明显下降而其他各组间则无显著变化. 结论此月经模型的子宫内膜血管形态学呈分层改变类似于人类子宫内膜的功能层和基底层.血管内皮细胞在子宫组织所占体积百分比仅在子宫内膜修复早期的实验组出现下降.

Objective： To study the cyclic changes of endometrial vasculature morphology and the volume fractions of endothelial cells with a murine menstruation model. Methods： Thirty-four adult ovariectomized C57BL/6 mice were used. After sequence hormone treatment, 0.2 ml arachis oil was injected into the uterine lumen to induce the decidual response, and the mice in control group were sham-operated without oil injection. Uteri of different groups were collected individually. Routine H＆E staining was carried on paraffin-embedded sections. The vasculature was displayed by immunohistochemistry with anti-CD31 monoclonal antibody. The computer image analysis software CAST2 was used to measure the volume fractions of endothelial cells. Results： H＆E staining displayed the endometrial sloughing at 30-35 hrs after oil treatment. At this time point, the endometrial vasculature showed two zones with immunohistochemistry. In the outer zone which was near the myometrium, vessels were evenly distributed while the vessels in the inner zone messed under the subluminal surface. In control group, endometrium remained intact and the whole layer of endometrial vasculature was disorganized. After oil treatment for 56-70 hrs, the epithelium was re-constructed in the experimental group, but the vessels were sparse and disorganized. In control group, the epithelium and vessels all appeared intact. Volume fraction of endothelial cell strikingly decreased only at 56-70 hrs after oil injection. Conclusions： The endometrial vasculature of this model showed two zones, which were morphologically similar to human endometrial functionalis and basalis. Volume fraction of endothelial cell only decreased after oil treatment for 56-70 hrs, whereas other groups did not show significant difference.