骨桥蛋白在肝素结合表皮生长因子样生长因子介导的小鼠胚胎对子宫内膜黏附和扩展中的作用研究

Role of osteopontin in heparin binding epidermal growth factor like growth factor induced embryo attachment and outgrowth in mouse endometrial cells

目的:探讨肝素结合表皮生长因子样生长因子(HB-EGF)对小鼠胚胎黏附和扩展中的作用机制。方法:建立小鼠胚胎与小鼠子宫内膜细胞共培养模型,分A、B、C、D、E共5组,分别加入不同浓度的HB-EGF(0μg/L、20μg/L、50μg/L、100μg/L、200μg/L),于倒置显微镜下统计各组胚胎黏附率及扩展率;用免疫组织化学法检测小鼠子宫内膜细胞骨桥蛋白(OPN)的表达及定位,Western blotting法测定各浓度HB-EGF对子宫内膜细胞OPN表达水平的影响,并分析其与HB-EGF的关联。结果:OPN在小鼠子宫内膜细胞高表达,主要分布于胞质中,随HB-EGF浓度升高,小鼠子宫内膜细胞中OPN的表达增强,小鼠胚胎对子宫内膜的黏附和扩展率也逐渐增高。结论:HBEGF可能通过上调小鼠子宫内膜细胞OPN的表达,促进小鼠胚胎对子宫内膜的黏附和扩展。

Objective： To investigate the mechanism of heparin binding epidermal growth factor like growth factor （HB-EGF） on the attachment and outgrowth of mouse embryos. Methods： An in vitro co-culture system of mouse embryos and mouse endometrial cells was established and then was divided into groups A, B, C, D, E with different concentrations of HB-EGF （0μg/L, 20 μg/L, 50 μg/L, 100 μg/L, 200 μg/L, respectively）. Blastoeyst attachment and outgrowth rates were then counted under the inverted microscope. Immunocytoehemical staining was used to detect the expression and localization of osteopontin （OPN） in mouse endometrial cells. Western blotting was used to detect the expression of OPN in different concentrations of HB-EGF, and the connective relations between them were analyzed. Results： OPN was highly expressed in mouse endometrial cells, mainly in cytoplasm; the expression of OPN in mouse endometrial cells was enhanced with the increase of HB-EGF concentrations, the blastocyst attachment and outgrowth rates were also gradually increased. Conclusion： By upregulating the expression of OPN in mouse endometrial cells, HB-EGF might promote the mouse embryo implantation.