申可佳++申奏秦旋++刘碧源++付灵梅+++熊桀

[摘要] 目的 探讨慢性制动应激建立卵巢早衰及卵巢低反应小鼠模型的方法。 方法 利用自制制动器对SPF级ICR雌性小鼠进行制动，造成慢性应激状态，观察小鼠的一般情况，称量体重，阴道涂片观察动情周期。制动完全结束后进行微刺激超排卵，在超排卵第3天和HCG注射后24 h计数各级卵泡。 结果 与未制动小鼠相比，制动期间小鼠的进食、毛色等一般情况变差，体重明显减轻（P < 0.05），动情周期异常的例数明显增多（P < 0.05），但超排卵第3天和HCG后24 h小鼠各级卵泡计数比较，制动小鼠与未制动小鼠没有明显区别（P > 0.05）。 结论 慢性制动应激可以引起小鼠卵巢功能紊乱或减退，但能否造成卵巢早衰及卵巢低反应还需进一步探讨。

[关键词] 卵巢早衰；卵巢低反应；慢性应激；小鼠

[中图分类号] R285.5 [文献标识码] A [文章编号] 1673-7210（2017）06（b）-0019-04

Study on establishment of premature ovarian function and poor ovarian response mice model through chronic stress

SHEN Kejia SHEN Zouqinxuan LIU Biyuan FU Lingmei XIONG Jie

TCM University of Hu'nan， Hu'nan Province， Changsha 410208， China

[Abstract] Objective To explore the method of chronic immobilization stress which can establish the mice model of premature ovarian function （POF） and poor ovarian response （POR）. Methods Self-made brakes were used to make chronic stress stateon SPFICR female mice， during the general situation， body weight and estrous cycle of these mice were observed. The microstimulated super ovulation was used on the mice after immobilization， and the follicles of the mice were counted respectively on the third day of superovulation and 24 h after HCG injection. Results Compared with those in the non-immobilization mice， the general situation such as eating， hair color and so on were worse， the body weight significantly reduced （P < 0.05）， and the number of abnormal estrous cycle distinct increased （P < 0.05） in the immobilization mice during the braking period. But there was no obvious difference of the follicle number between the immobilization mice and the non-immobilization mice at various levels on the third day of superovulation and 24 h after HCG injection （P > 0.05）. Conclusion Chronic immobilization stress can causethe disorder or degradation of ovarian function，and further study is needed to find whetherit can cause premature ovarian function and poor ovarian response.

[Key words] Premature ovarian function； Poor ovarian response； Chronic stress； Mice

輔助生殖超排卵过程中卵巢对促性腺激素的刺激反应低下称为卵巢低反应（POR），病因与年龄、遗传和免疫、手术和感染、环境等因素相关，还有一类无明确病因可循，可能是因长期压力应激导致卵巢功能衰退（特发性POF）而引起的POR在临床也越发多见[1-4]。目前基础研究大多为化疗药物、免疫及半乳糖等所致POF动物模型，少见特发性POF/POR动物模型[5-9]。本实验拟在模拟人类特发性POF的发病经过，用重复制动造成小鼠慢性应激状态，并在制动结束后进行超排卵，探讨是否能建立POF/POR小鼠模型。

1 材料与方法

1.1 实验材料

1.1.1 实验动物

6周龄健康SPF级ICR雌性小鼠80只和成年雄性小鼠20只，体重为22～25 g，由湖南斯莱克景达实验动物有限公司提供，动物合格证号：SCXK（湘）2011-0003。实验遵循湖南省实验动物保护和使用指南，并经湖南中医药大学实验动物伦理委员会批准。……