Asoprisnil抗小鼠孕卵着床效果及对子宫内膜容受性的影响

A Study of the Anti-implantation Effect of Asoprisnil and Its Effect on Endometrial Receptivity in Mice

目的：探讨Asoprisnil抗小鼠孕卵着床的效果及其对小鼠着床窗期子宫内膜容受性的影响。方法：将孕第1日小鼠随机分成4组，分别为Asoprisnil低（5og／g）、中（10vg／g）、高（20μg／g）剂量组和对照组，每组22只，于孕第1～3日每日灌胃给药1次，对照组以体积分数1％羧甲基纤维素钠替代，第5日每组处死孕鼠5只取子宫组织，HE染色观察子宫内膜形态学改变，免疫组织化学s．P法检测子宫内膜PCNA表达。第8日处死余下的孕鼠，计数着床点数。结果：①Asoprisnil高、中和低剂量组的妊娠率分别为11．76％、35．29％和76．47％，与对照组（94．12％）比较差异有统计意义俨〈0．001）。②Asoprisnil高、中、低剂量组着床胚泡数第50百分位数（PSO）分别为13（10．5~14）、10（0．5～11）和0（0~12），与对照组0（0～0）比较差异均有统计学意义（P〈0．001）。③对照组围着床期子宫内膜腺体弯曲折叠，为复层高柱状上皮细胞；基质细胞蜕膜变，细胞大且排列疏松，胞质丰富透亮；内膜中腺体和血管丰富。Asoprisnil组子宫内膜腺体为单层或者复层上皮细胞；内膜基质细胞蜕膜变不明显，基质细胞较小，排列致密；腺体和血管增生不明显。④围着床期小鼠子宫内膜腺体和基质中均有PCNA表达。内膜腺体中Asoprisnil高、中、低剂量组PCNA表达强度（AIOD）分别为0．15±0．01、0．16±0．03和0．14±0．02，与对照组（0．21±0．03）比较差异有统计学意义（P〈0．001）。Asoprisnil高、中、低剂量组内膜基质细胞中PCNA表达强度（AIOD）分别为0．17±0．01、0．18±0．03和0．17±0．02，与对照组（O．15±0．02）差异有统计学意义（P〈0．001）。结论：Asoprisnil能显著抑制着床窗期子宫内膜腺体增殖，促进子宫内膜基质细胞增殖，但阻止基质细胞蜕膜化，降低子宫内膜容受性而发挥抗小鼠胚胎着床效应，显示出潜在的子宫内膜靶向避孕前景。

Objective： To investigate the effectiveness of asoprisnil in interrupting the implantation and its effect on endometrial receptivity in implantation window period in mice. Methods： Post-coitus mice were administered with different doses of asoprisnil [high dose （20 μg/g） group, medium dose （10 μg/g） group and low dose （5 μg/g） group] or 1% carboxymethyl cellulose （the control） p.o. for the first 3 d of pregnancy. HE staining was used to survey the morphology changes of endometrium in the implantation window period （5th d of pregnant, 5 mice per group）. At the same time, immunohistochemistry （IHC） was then used to test the expression of PCNA in endometrium. Some mice （17 mice per group） sacrificed on the 8th day of pregnancy and the uterine horns were examined for the presence or absence 0fnidation sites and the number of implantation embryos. Results： 1） Pregnancy rates in high dose group, medium dose group and low dose group were 11.76%, 35.29% and 76.47%, respectively, and the difference was statistically significant compared with the control （P〈0.001）. 2） The median number （P50） of implantation embryo in control group, low dose group, medium dose group and high dose groups was respectly as 13 （10.5-14）, 10 （0.5-11）, 0 （0-12） and 0 （0-0）, and the differences were statistically significant compared with the control group （P〈0.001）. 3） In the control, endometrium in implantation window period was characterized by luminal gland proliferation with multiple tall and columnar surface epithelial cells, vascular congestion and equality scattered among the decidual cells which was big and with abundant matrix. While in experimental groups, the gland proliferation was not significant, the luminal epithelial cells were single and multiple layers distributed irregularly. Stromal cells were compact and small, without decidual transformation. Glandular and vascular was infrequent in endometrium. 4） IHC indicated that PCNA was expressed both in luminal epithelium cells and stromal cells. In luminal epithelium cells, the AIOD in high, medium, low dose groups and the control were 0.15 ± 0.01, 0.16 ± 0.03, 0.14 ± 0.02 and 0.21 ± 0.03, respectively, the difference was statistically significant in asoprisnil treated groups compared with the control （P〈0.001）. While in stromal cells, the AIOD in high, medium, low dose groups and the control were 0.17 ±0.01, 0.18 ± 0.03, 0.17 ± 0.02 and 0.15 ±0.02, respectively, and the difference was statistically significant （P〈0.001）. Conclusion： Asoprisnil can effectively disturb embryo implantation in mice. Asoprisnil suppressed the proliferative of luminal epithelium by decreasing the expression of PCNA, while promot- ing the proliferation of the stromal cells. The endometrium development was asynchronous, so endometrial recep- tivity was affected and finally resulted in embryo implantation failure. Asoprisnil was a good candidate for endome- trial targeted oral contraceptive compound.