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目的选用雌性昆明小鼠建立慢性轻度不可预见性应激(CUMS)动物模型,了解CUMS对雌性昆明小鼠生殖功能的影响,为后续探讨应激对生殖功能的影响机制提供适宜的动物模型。方法将240只雌性昆明小鼠以正常饲养5 d后,采用随机数字表法分为对照组(90只)和实验组(150只)。雄性小鼠(为合笼备用)、对照组小鼠均按正常条件饲养。将实验组150只雌性昆明小鼠以9种慢性、轻度和不可预见性应激因子在4周内随机应用造模。4周后,对两组小鼠采用旷场实验和糖水消耗实验,观察体质量变化,以进行有效性验证。对造模成功的CUMS小鼠进行孕马血清促进腺激素(PMSG)/人绒毛膜促性腺激素(HCG)促排卵,通过获取卵巢组织标本、卵母细胞、2-细胞期胚胎、孕4 d胚胎(囊胚、桑葚胚),并进行孕5 d体内着床位点实验,以观察卵巢反应及胚胎发育潜能。结果 4周CUMS刺激后,与对照组小鼠相比,实验组小鼠体质量增加、2%糖水消耗实验及旷场实验均存在明显差异(P<0.05)。PMSG/HCG促排治疗后,实验组小鼠卵巢的窦卵泡及排卵前卵泡较对照组明显减少(P<0.05);实验组小鼠的卵母细胞数、受精率、2-细胞胚胎数、孕4 d胚胎数、囊胚数、优胚率及孕5 d着床位点数均明显低于对照组,差异均有统计学意义(P<0.05)。结论 CUMS雌性昆明小鼠模型是一种生殖功能减退的情绪应激动物模型,具有有效性、可操作性和可重复性,可进一步用于生殖医学领域机制方面的研究。
Objective To establish a chronic mild unpredictable stress (CUMS) animal model in female Kunming mice to understand the effect of CUMS on reproductive function in female Kunming mice and to provide a suitable animal model for the subsequent study of the mechanism of stress on reproductive function. Methods Two hundred and four female Kunming mice were fed for 5 days and divided into control group (n = 90) and experimental group (n = 150) by random number table. Male mice (for cage reserve), control mice were fed under normal conditions. 150 female Kunming mice in the experimental group were randomized to model with nine chronic, mild and unpredictable stress factors within 4 weeks. After 4 weeks, open-field test and sugar consumption test were performed on the two groups of mice to observe the change of body weight for validation. Ovarian tissue samples, oocytes, 2-cell stage embryos, pregnant 4-day embryos were harvested from CUMS mice that were successfully treated with PMSG and human chorionic gonadotropin (HCG) Embryos (blastocyst, morula), and in vivo 5 days pregnant implantation site experiments to observe the ovarian response and embryonic development potential. Results After CUMS stimulation for 4 weeks, compared with the control group, the mice in the experimental group showed an increase in body weight, a significant difference (P <0.05) in the 2% sugar water consumption experiment and the open field test. PMSG / HCG promoted the ovarian antral follicles and preovulatory follicles in the experimental group significantly decreased compared with the control group (P <0.05); the number of oocytes in the experimental group, fertilization rate, the number of 2-cell embryos, The number of embryos, blastocyst number, excellent embryo rate and the number of implantation sites at 5 days of pregnancy on day 4 of pregnancy were significantly lower than those of the control group (P <0.05). CONCLUSION CUMS female Kunming mouse model is an animal model of reproductive failure induced by emotional stress, which has the characteristics of validity, maneuverability and repeatability and can be further applied in the field of reproductive medicine.