小鼠月经生理模型的探讨

Seeking an Optimized Murine Menstruation Model

目的减少Finn CA于1984年首次报道的小鼠月经模型的观察时间点,以期为月经生理学研究提供一种较廉价且易操作的月经模型。方法应用成年雌性去势C57BL/6小鼠,给予续贯性激素处理,最末次激素处理后4～6h,实验组小鼠宫腔内注射花生油以诱导子宫内膜蜕膜化反应,对照组小鼠给予同样激素处理但无宫腔油剂注射。分别于油剂处理后31～35h(T3组)、56～70h(T4组)处死小鼠,称量子宫湿重,制作H&E组织切片,运用图像分析软件CAST2,计算全子宫横截面积(TUA)与子宫内膜横截面积(EA)。结果H&E染色子宫组织切片示在单纯雌激素作用下宫内膜呈单层立方上皮,核浆比较高,内膜基质疏松;雌孕激素联合处理后,分泌细胞易见,腺腔内可见分泌物。激素撤退后实验组T3观察到子宫内膜剥离,T4组示子宫内膜修复。对照组子宫内膜始终完整。子宫湿重在激素撤退后,实验组下降较慢。激素撤退后实验组T3的TUA继续上升而EA则维持原水平,T4组TUA与EA均明显下降。结论此模型在子宫内膜剥落期和早期修复期组织学特征与人类子宫内膜有一定的相似性。

Objective This study tried to reduce the observation time points in the establishment of a murine menstruation model, which was originally developed by Finn CA in 1984. Therefore, to make the murine menstruation model more economical and easier to handle. Methods Adult female C57BL/6 mice were ovariectomized, then sequentially treated with hormones. 4- 6 hours after the last hormone injection, 0.02 ml sterilized arachis oil was injected into the uterine lumen of experimental mice. The control group only received the sham operation. Uteri were collected after oil treatment for 31-35 hours （group T3） and 56-70 hours （group T4）, measuring wet uterine weight, doing H＆E staining and using CAST2 image analysis software to measure the total uterine cross-sectional area （TUA） and endometrial cross-sectional area （EA）. Results Through histological examination, estrogen treated group showed simple columnar epithelial cells, with high nucleus-to-cytoplasm ratio and elongate sausage-shaped nuclei with dense chromatin. After combined treatment with progesterone, secretory cells and their secretions were easily found in the glandular lumens. After oil treatment endometrium sloughing was observed in group T3, while endometrial epithelium repaired completely in group T4. Endometrium of control groups kept intact during the whole process. Wet uterine weight had no significant changes during sequential hormone treatment. After hormone withdrawal, the wet uterine weight decreased in the experimental groups slower than that of control mice. TUA and EA significantly increased after combined treatment with progesterone. 30-35 hours after oil treatment TUA increased, whereas EA did not changed. 56-70 hours later both of them decreased. Conclusion The histological changes of endometrium in this murine menstruation model displayed many features resembling human menstruation, especially, the endometrial subluminal epithelium sloughing and early endometrial repair.