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p16~(INK4a)在早孕小鼠子宫内膜的表达及其对胚泡着床的影响

杨环, 谢怡, 杨戎, 魏莎莉, 郗强

重庆医科大学生殖生理教研室.重庆 400016

摘要

该研究旨在检测肿瘤抑制基因p16~(INK4a)(inhibitor of cyclin--dependent kinase4a)在早孕小鼠子宫内膜中的表达规律,探讨p16~(INK4a)在小鼠胚胎着床过程中的作用。采用荧光定量PCR(FQ--PCR)和免疫组织化学方法分别检测未孕小鼠及孕小鼠d2、d3、d4、d5、d7子宫内膜p16~(INK4a) mRNA和蛋白的表达;子宫角注射p16~(INK4a)抗体观察胚泡着床数。FQ--PCR结果显示孕小鼠子宫内膜组织p16~(INK4a) mRNA的表达高于未孕小鼠,且随着妊娠天数的增加呈现表达逐渐增强的趋势,到妊娠d5达到最高,后渐降。免疫组织化学分析显示p16~(INK4a)蛋白在子宫内膜的表达规律与mRNA结果一致。子宫角注射p16~(INK4a)抗体后胚泡着床数明显减少。以上结果提示,p16~(INK4a)在妊娠早期子宫内膜持续表达,可能参与胚泡着床。

关键词： 胚胎着床; 子宫内膜; p16~(INK4a); 实时荧光定量PCR; 免疫组织化学; 体内实验

Expression of p16~(INK4a) in mouse endometrium and its effect during blastocyst implantation

Yang Huan, Xie Yi, Yang Rong, Wei Shali, Xi Qiang

Department of Reproductive Physiology,Chongqing Medical University.Chongqing 400016

Abstract

The expression of tumor suppressor gene p16~(INK4a) in mouse endometrium during early pregnancy and its possible role in blastocyst implantation were investigated in the present study. Real-time fluorescent quantitative PCR (FQ-PCR) and immunohisto- chemistry were applied to detect p16~(INK4a) mRNA and protein expressions in endometrium of un-pregnant and pregnant mice on day 2, 3, 4, 5, 7, respectively. In addition, p16~(INK4a) antibody was injected into the horns of uteri in pregnant mice on day 3 and its effect during blastocyst implantation was detected in vivo. The higher expressions of p16~(INK4a) mRNA and protein were observed in pregnant mice compared with that in un-pregnant mice, with a steady increase from day 2 to day 5 and reaching the maximal level on day 5 of pregnancy and then decreasing. p16~(INK4a) antibody decreased the number of implanted blastocysts compared with that of saline-injected group. The results suggest that p16~(INK4a) may be associated with apoptosis of luminal epithelial cells and decidual cells, coordinating decidualization of endometrium and invasion of trophoblastic cells. Thus, we presume that p16~(INK4a) participates in the process of blastocyst implantation in mice.

Key words: blastocyst implantation;Endometrium;p16~(INK4a);real-time fluorescent quantitative PCR;Immunohistochemistry;in vivo

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引用本文：

杨环, 谢怡, 杨戎, 魏莎莉, 郗强. p16~(INK4a)在早孕小鼠子宫内膜的表达及其对胚泡着床的影响[J]. 生理学报 2008; 60 (4): 547-552.

Yang Huan, Xie Yi, Yang Rong, Wei Shali, Xi Qiang. Expression of p16~(INK4a) in mouse endometrium and its effect during blastocyst implantation. Acta Physiol Sin 2008; 60 (4): 547-552 (in Chinese with English abstract).