GRB7基因在早孕小鼠子宫内膜中的表达

Expression of growth factor receptor bound 7 in the endometrium of early pregnant mouse

摘要:

目的：研究生长因子结合受体7（growth factor receptor bound 7，GRB7）在早孕小鼠子宫内膜中的表达规律，探讨其在胚胎着床过程中的作用。方法：分别应用实时荧光定量PCR、免疫组化和Western blot方法检测未妊娠及妊娠早期d1～7小鼠子宫内膜GRB7基因及蛋白的表达；孕3 d下午8点子宫角注射GRB7反义寡核苷酸，孕7 d观察胚泡着床数较正常组有无变化。结果：实时荧光定量PCR测得未妊娠（d0）和妊娠早期（d1～7）小鼠子宫内膜组织均有GRB7 mRNA表达，并在孕5 d达到峰值；免疫组化及Western blot分析显示GRB7蛋白在子宫内膜的表达规律与实时荧光定量PCR基本一致，孕1 d到孕4 d，GRB7蛋白主要表达部位为腔上皮、腺上皮，基质细胞少量表达，孕5 d、孕6 d，GRB7蛋白表达较之前增强，并且在基质细胞开始明显表达，孕5 d最强；孕6 d着床点GRB7蛋白高表达于初级蜕膜区，着床旁GRB7蛋白高表达于基质细胞；子宫角注射GRB7反义寡核苷酸后胚泡着床数较正常组明显减少。结论：GRB7在早孕小鼠子宫内膜中呈动态表达，提示它参与了胚胎着床的发育过程。

Abstract:

Objective：To explore the role of growth factor receptor bound 7（GRB7） during embryo implantation through detecting its expression pattern in the mouse endometrium. Methods：Expression of GRB7 mRNA and protein in the non-pregnant and pregnant mouse endometrium from d1 to d7 was examined by qRT-PCR，immunohistochemistry and Western blot. GRB7 antisense oligo-nu－cleotide was injected at 8 pm on pregnancy d3 and embryo implantation number on pregnancy d7 was examined. Results：GRB7 mR－NA expressed in uterine endometrium of the non-pregnant and pregnant mouse from d1 to d7 and reached the highest level on preg－nant d5 based on the results of qRT-PCR. Immunohistochemistry and Western blot showed that the expression pattern of GRB7 in mouse endometrium was the same with that detected by qRT-PCR. Moreover，from pregnant d1 to d4，GRB7 was mainly expressed in epithelium and glandular cells and was lowly expressed in stromal cells；on d5 and d6，GRB7 was more intense in epithelium and glandular cells and was obviously expressed in stromal cells，especially on d5. On d6，GRB7 was expressed strongly in primary decid－ual zones at implantation sites and stromal cells at inter-implantation sites，respectively. After horn injecting GRB7 antisense oligonu－cleotides，the number of embryo implantation was obviously decreased when compared with that in control group. Conclusions：Differ－ential expression of GRB7 in pregnant mouse endometrium suggests that GRB7 may involve in the process of embryo implantation.