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中华妇幼临床医学杂志(电子版) ›› 2017, Vol. 13 ›› Issue (04) : 419 -424. doi: 10.3877/cma.j.issn.1673-5250.2017.04.009

所属专题: 文献

论著

小鼠流产模型的建立及其蜕膜组织中Yes相关蛋白表达初探
陈恒禧1, 傅璟1, 周敏2, 黄薇1,()   
  1. 1. 610041 成都,四川大学华西第二医院妇产科、出生缺陷与相关妇儿疾病教育部重点实验室
    2. 610072 成都,四川省人民医院辅助生殖医学中心
  • 收稿日期:2017-04-11 修回日期:2017-07-01 出版日期:2017-08-01
  • 通信作者: 黄薇

Study on the establishment of mice abortion model and the expression of Yes-associated protein in decidual tissues

Hengxi Chen1, Jing Fu1, Min Zhou2, Wei Huang1,()   

  1. 1. Department of Gynecology and Obstetrics, Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China
    2. Reproductive Medicine Center, Sichuan Provincial People′s Hospital, Chengdu 610072, Sichuan Province, China
  • Received:2017-04-11 Revised:2017-07-01 Published:2017-08-01
  • Corresponding author: Wei Huang
  • About author:
    Corresponding author: Huang Wei, Email:
引用本文:

陈恒禧, 傅璟, 周敏, 黄薇. 小鼠流产模型的建立及其蜕膜组织中Yes相关蛋白表达初探[J]. 中华妇幼临床医学杂志(电子版), 2017, 13(04): 419-424.

Hengxi Chen, Jing Fu, Min Zhou, Wei Huang. Study on the establishment of mice abortion model and the expression of Yes-associated protein in decidual tissues[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2017, 13(04): 419-424.

目的

探讨Yes相关蛋白(YAP)在蜕膜组织中的表达强度与小鼠流产的关系。

方法

2013年6月至2015年1月,选择无特殊病原体(SPF)级21只雌性CBA/J小鼠、5只雌性DBA/2J小鼠和6只雄性BLAB/c小鼠为研究对象。21只雌性CBA/J小鼠按照随机数字表法,随机分成2组,将10只雌性CBA/J小鼠与5只雄性DBA/2J小鼠合笼,建立雌性(♀)CBA/J×雄性(♂)DBA/2J流产模型,纳入流产组,将11只雌性CBA/J小鼠与6只雄性BALB/c小鼠合笼,建立♀CBA/J×♂BALB/c正常妊娠模型,纳入对照组。记录2组雌性CBA/J小鼠存活胚胎数及流产率,随后采用苏木精-伊红(HE)染色观察蜕膜组织,并采用免疫组织化学染色法检测YAP在蜕膜组织中的分布,以及YAP表达强度。采用统计学方法比较2组小鼠存活胚胎数、流产率及不同YAP表达强度的小鼠数。实验过程中对动物的处置符合动物伦理学要求。

结果

①流产组小鼠存活胚胎数的中位数(5.0个)低于对照组小鼠存活胚胎数的中位数(7.5个),2组比较,差异有统计学意义(Z=-2.678,P=0.007)。流产组小鼠流产率的中位数(33.3%)高于对照组小鼠流产率的中位数(5.6%),2组比较,差异亦有统计学意义(Z=-3.797,P<0.001)。②YAP主要表达于小鼠蜕膜细胞的细胞质中,将免疫组织化学结果进行半定量评分的结果显示,流产组YAP表达水平较对照组降低,并且差异有统计学意义(Z=-2.445,P=0.014)。

结论

♀CBA/J×♂DBA/2J小鼠流产模型中的胚胎丢失具有高度重复性,流产率较高且相对恒定。YAP可能参与流产的发生、发展。

Objective

To establish animal abortion model and investigate the relation between Yes-associated protein(YAP) expression and abortion.

Methods

From June 2013 to January 2015, 21 female CBA/J mice, 5 male DBA/2J mice and 6 male BALB/c mice were chosen as research subjects. Twenty-one female CBA/J mice were randomly divided into 2 groups, 10 female CBA/J mice mated with BALB/c to establish abortion model as abortion group (♀CBA/J×♂DBA/2J), and another 11 female CBA/J mice mated with 6 male BALB/C mice to establish control group (♀CBA/J×♂BALB/c). The numbers of survival embryo and abortion rates between abortion group and control group were recorded. The expression of YAP was examined by immunohistochemistry in decidua tissues and was semi-quantitatively assessed of abortion group and control group. All experimental protocols were approved by the Institutional Animal Care and Use Committee.

Results

①The median number of survival embryo in abortion group (5.0) was lower than that in control group (7.5), and the difference was statistically significant(Z=-2.678, P=0.007). The median abortion rate of abortion group (33.3%) was higher than that of control group (5.6%), and the difference was statistically significant (Z=-3.797, P<0.001). ②YAP was mainly expressed in the cytoplasm of decidua cells. Semi-quantitative immunohistochemistry showed that the YAP expression in abortion group was lower than that in control group, and the difference was statistically significant (Z=-2.445, P=0.014).

Conclusions

Embryo loss is highly repetitive in ♀CBA/J×♂DBA/2J abortion model and the abortion rate is high and relatively constant. It is suggested that YAP might be involved in the occurrence and development of abortion.

表1 流产组小鼠合笼后妊娠情况
表2 对照组小鼠合笼后妊娠情况
图1 2组小鼠妊娠蜕膜组织染色结果(图1A:流产组;图1B:对照组)(HE染色,高倍镜)
表3 2组小鼠蜕膜组织不同Yes相关蛋白表达强度的小鼠数(只)
图2 YAP在蜕膜组织中的不同表达强度(图2A:流产组低表达;图2B:流产组中表达;图2C:流产组高表达;图2D:对照组低表达;图2E:对照组中表达;图2F:对照组高表达) (免疫组织化学SP法染色,高倍镜)
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