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促性腺激素释放激素激动剂控制性超促排卵对小鼠胚胎着床的作用及其机制

吴琼, 徐华洲, 杨江霞, 李志丽, 傅天, 杜惠兰, 宋翠淼^*

河北中医学院，中西医结合生殖疾病协同创新中心，河北省中西医结合肝肾病证重点实验室，石家庄 050200

摘要

本研究旨在探讨促性腺激素释放激素激动剂(gonadotropin-releasing hormone agonist, GnRHa)控制性超促排卵(controlled ovarian hyperstimulation, COH)对小鼠胚胎着床的作用及机制。40只9周龄昆明雌性小鼠随机分为两组：COH组腹腔注射GnRHa醋酸丙氨瑞林+人绝经期促性腺激素(human menopausal gonadotropin, HMG)+人绒毛膜促性腺激素(human chorionic gonadotrophin, hCG)，对照组腹腔注射等体积生理盐水。注射hCG日16:00雌雄小鼠合笼，妊娠第4天每组处死10只小鼠，用放射免疫法检测血清雌二醇(estradiol, E2)、孕酮(progestrone, P)的水平；用HE染色观察卵巢、子宫内膜组织形态；用Western blot和免疫组织化学法检测子宫内膜白血病抑制因子(leukemia inhibitory factor, LIF)、磷酸化信号转导和转录活化因子3 (phosphorylated signal transducer and activator of transcription 3, p-STAT3)、肝素结合表皮生长因子样生长因子(heparin-binding epidermal growth factor-like growth factor, HB-EGF)和glycodelin A蛋白的变化。妊娠第8天每组处死10只小鼠，观察胚胎着床情况。结果显示，与对照组比较，COH组小鼠妊娠第4天(胚胎着床期)血清E2水平降低(P < 0.05)，P水平升高(P < 0.05)；卵巢组织各级卵泡少见，可见多个黄体，子宫内膜变薄，腺体数量减少(P < 0.05)；子宫内膜LIF、p-STAT3、HB-EGF和glycodelin A蛋白表达下降(P < 0.05)；妊娠第8天胚泡发育缓慢，数量减少(P < 0.05)。以上结果提示，GnRHa COH可影响小鼠胚胎着床，其机制可能与着床期内源性激素失衡，子宫内膜结构和LIF、p-STAT3、HB-EGF、glycodelin A蛋白表达改变，导致子宫内膜容受性降低、胚胎-子宫对话异常有关。

关键词： 促性腺激素释放激素激动剂; 控制性超促排卵; 胚胎着床

分类号：Q492.6

Effect of GnRHa controlled ovarian hyperstimulation on mouse embryo implantation and its mechanism 

WU Qiong, XU Hua-Zhou, YANG Jiang-Xia, LI Zhi-Li, FU Tian, DU Hui-Lan, SONG Cui-Miao^*

Hebei University of Chinese Medicine, Collaborative Innovation Center of Integrated Chinese and Western Medicine on Reproductive Disease, Hebei Key Laboratory of Integrative Medicine on Liver-kidney Patterns, Shijiazhuang 050200, China

Abstract

The purpose of the present study was to investigate the effects and underlying mechanism of gonadotropin-releasing hormone agonist (GnRHa) controlled ovarian hyperstimulation (COH) on embryo implantation in mice. Forty female Kunming mice aged 9 weeks were randomly divided into two groups (control and COH groups). The COH group received intraperitoneal (i.p.) injections of aminocyclin acetate (GnRHa), human menopausal gonadotropin (HMG) and human chorionic gonadotropin (hCG), while the control group was given equal amount of physiological saline by i.p. injection. One male mouse and two female mice were put into the same cage at 16:00 on the hCG injection day, and on the fourth day of pregnancy, 10 mice from each group were killed. The levels of serum estradiol (E2) and progesterone (P) were measured by radioimmunoassay; HE staining was used to observe the morphology of ovarian and endometrial tissues. The protein expression levels of endometrial leukemia inhibitory factor (LIF), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), heparin-binding epidermal growth factor-like growth factor (HB-EGF) and glycodelin A were detected by Western blot and immunohistochemistry. Ten mice from each group were sacrificed on the eighth day of pregnancy, and the status of the uterus and the average number of blastocysts were observed. The results showed that, compared with control group, the serum E2 level in COH group was significantly decreased (P < 0.05), while the P level was increased significantly (P < 0.05); the ovarian follicles at different developmental stages were rare, corpus lutea (CL) were visible and multiple, the endometrium was thinned, and the number of endometrial glands was reduced (P < 0.05); the contents of LIF, p-STAT3, HB-EGF and glycodelin A in the endometrium were decreased significantly (P < 0.05) on the fourth day of pregnancy; mouse blastocysts developed slowly and were decreased in number on the eighth day of pregnancy (P < 0.05). The above results suggest that GnRHa COH can affect embryo implantation in mice. The mechanism may be related to the imbalance of gonadal hormone, the changes in the structure of the endometrium and the expressions of LIF, p-STAT3, HB-EGF and glycodelin A in the implantation stage, which may lead to the decrease of endometrial receptivity and the abnormal dialogue between the embryo and the uterus.

Key words: GnRHa; controlled ovarian hyperstimulation; embryo implantation

收稿日期：2018-06-20　　录用日期：2018-10-12

通讯作者：宋翠淼　　E-mail: songcuimiao@126.com

DOI: 10.13294/j.aps.2018.0067

引用本文：

吴琼, 徐华洲, 杨江霞, 李志丽, 傅天, 杜惠兰, 宋翠淼. 促性腺激素释放激素激动剂控制性超促排卵对小鼠胚胎着床的作用及其机制[J]. 生理学报 2018; 70 (5): 489-496.

WU Qiong, XU Hua-Zhou, YANG Jiang-Xia, LI Zhi-Li, FU Tian, DU Hui-Lan, SONG Cui-Miao. Effect of GnRHa controlled ovarian hyperstimulation on mouse embryo implantation and its mechanism . Acta Physiol Sin 2018; 70 (5): 489-496 (in Chinese with English abstract).