慢性束缚应激对围着床期小鼠胚泡的影响及其机制  

Impact of Chronic Restraint Stress on Embryos during Peri-Implantation Period in Mice

作　　者：刘红宇 叶婵琦 刘仁平 黄博抒 胡炜华 王晶磊[1] LIU Hong-yu;YE Chan-qi;LIU Ren-ping;HUANG Bo-shu;HU Wei-hua;WANG Jing-lei(Department of Physiology,School of Basic Medical,Nanchang University,Nanchang 330006,China;Periodical Press,Nanchang University,Nanchang 330006,China;Reproductive Genetics ResearchRoom,Pingxiang Maternal and Child Health Hospital,Pingxiang 337000,China)

机构地区：[1]南昌大学基础医学院生理学教研室,南昌330006 [2]萍乡市妇幼保健院生殖遗传研究室,江西萍乡337000 [3]南昌大学期刊社,南昌330006

出　　处：《南昌大学学报（医学版）》2018年第5期6-11,共6页Journal of Nanchang University：Medical Sciences

基　　金：江西省自然科学基金(20142BAB205001)

摘　　要：目的探讨慢性束缚应激刺激对围着床期小鼠模型胚泡着床的影响及其机制。方法选择10～12周龄的健康、清洁级性成熟的雌性和雄性昆明种小鼠各20只,建立妊娠动物模型。将16只孕鼠随机分为应激组和对照组,每组8只。应激组小鼠在妊娠第3天(D3)上午开始给予束缚应激刺激,连续3d。对照组小鼠在应激组束缚应激刺激时给予断水、断食,同时不给予任何处理。妊娠D5慢性束缚应激刺激后,采用酶联免疫吸附法(ELISA法)检测血清皮质酮、孕酮和催乳素的水平,尾静脉注射0.5%锥虫兰染液观察小鼠子宫内膜组织胚泡的着床位点数,观察、测定孕鼠子宫蜕膜胚泡数及孕鼠体质量、子宫和卵巢质量,并计算子宫和卵巢指数,HE染色法观察慢性束缚应激刺激对小鼠子宫蜕膜组织间质细胞和腺体的病理学改变,半定量RT-PCR法检测子宫蜕膜组织中IL-10mRNA和TNF-αmRNA水平的变化。结果与对照组比较,应激组血清皮质酮水平明显升高,催乳素、孕酮水平均明显降低,子宫内膜胚泡着床位点数减少,体质量、子宫质量均减轻,子宫指数减少,光学显微镜观察显示对照组子宫蜕膜内胚泡呈串珠状,胚泡大小和形状基本一致,分布均匀。应激组子宫蜕膜见胚泡串珠状部分消失并分布不均匀,子宫蜕膜胚泡数减少。子宫蜕膜中IL-10mRNA、TNF-αmRNA表达水平均明显降低,TNF-αmRNA/IL-10mRNA比值明显升高(均P<0.05)。结论围着床期慢性束缚应激刺激可使血清中催乳素和孕酮的含量下降,皮质酮含量升高,并可能导致子宫局部Th1/Th2免疫平衡向Th1漂移,降低小鼠胚泡着床数,从而影响胚泡着床。Objective To discuss the effect of chronic restraint stress on implantation and development of mouse peri-implantation blastocysts and its mechanisms.Methods Kunming mice(10-12weeks old,20females and20males)were used to establish pregnant animal model.Sixteen pregnant mice were randomly divided into two groups,with8mice in each group.On the morning of day3of pregnancy(D3),mice in restraint stress group were subjected to restraint stress for3days.Meanwhile,mice in control group were fasted without other treatment.Enzyme-linked immunosorbent assay was used to detect corticosterone,prolactin and progesterone levels in peripheral blood of pregnant mice on D5.The number of implantation sites was observed by tail vein injection of0.5%trypan blue staining.The number of blastocysts in uterine decidua,body weight,uterus mass,ovary mass,uterus index,and ovary index were measured in pregnant mice.HE staining was utilized to observe the pathological changes of stromal cells and glands in uterine decidua.RT-PCR assay was used to investigate the mRNA levels of IL-10and TNF-αin uterine decidua.Results Compared with control group,chronic restraint stress significantly increased corticosterone level,decreased prolactin and progesterone levels,and reduced the number of implantation sites,body weight,uterus mass and uterus index.Optical microscopy showed beaded and evenly distributed blastocysts with basically same size and shape in uterine decidua in control group.However,the beaded structure partially disappeared with asymmetric distribution and the number of blastocysts decreased in uterine decidua in restraint stress group.In addition,restraint stress inhibited the mRNA expression of IL-10and TNF-αand increased the ratio of TNF-αmRNA to IL-10mRNA,compared with control group(P<0.05).Conclusion The restraint stress in peri-implantation can decrease the levels of prolactin and progesterone,increase the level of corticosterone,induce Th1/Th2balance drift toward Th1,reduce the number of embryo implantation sites,and thus negative

关 键 词：慢性束缚应激 围着床期 胚泡着床 子宫蜕膜 皮质酮 白细胞介素-10 肿瘤坏死因子-α 动物 实验 小鼠 

分 类 号：R392.12[医药卫生—免疫学] R-332[医药卫生—基础医学]