【作者】 周赛男；

【导师】 燕秋；

【作者基本信息】 大连医科大学 ， 生物化学与分子生物学， 2005， 硕士

【摘要】 目的:胚胎着床与多种因素协同作用相关,其中包括岩藻糖化的Lewis 寡糖。本实验室前期研究发现,小鼠子宫内膜 LeY 在妊娠第四天即着床窗口期表达最强,用 LeY 特异性抗体封闭子宫内膜表面的 LeY寡糖可明显的抑制胚泡的着床过程;临床不孕症患者的子宫内膜中 LeY的表达减少。提示 LeY抗原与着床密切相关,但其表达和调控机制需进一步探讨。表达于早期胚胎的阶段特异性胚胎抗原 LeX(stage specificembryonic antigen, SSEA)在早孕大鼠的子宫内膜腔上皮表达明显增加,但其功能尚未明了,推测与子宫内膜对胚胎的接受性变化有关。输卵管与卵巢在早孕过程中行使重要的功能,二者是否表达 LeY/LeX以及其表达的调控尚未见文献报道。本研究旨在了解 LeX/LeY在早孕小鼠子宫内膜与输卵管,及卵巢的表达和趋势,并进一步分析其合成关键酶——岩藻糖基转移酶 FUT9、FUT1 和 FUT4 的表达,探讨小鼠早孕期寡糖抗原的表达及调控与功能的关系。 方法:(1)用免疫组织化学方法检测妊娠 1-7 天小鼠子宫、输卵管与卵巢组织的 LeX/LeY抗原的表达。(2)用 RT-PCR 方法对合成 LeX关键酶 FUT9 与合成 LeY的关键酶 FUT1 和 FUT4 的基因进行分析。(3)用Western-blot对早孕小鼠卵巢所表达的LeY寡糖抗原结合糖蛋白进行检测。更多还原

【Abstract】 Objective Embryonic implantation involves in many factors includingfucosylated oligosaccharide. The study of our lab found that LeY on theendometriums was expressed highly on the fourth day after pregnancy, andendometriums which exposed to specific antibody to LeYcould be inhibitedin the process of implantation. Moreover LeY was expressed lowly in theendometriums of women who were sterile. All the facts suggested that thisantigen has a close relationship with implantation, but the function neededto be explored further. LeX that is the stage specific embryonic antigen(SSEA) is expressed on the early embryo and expressed highly in the earlypregnant rats’ endometriums, but the function is beyond known. Ovary andoviduct are of the parts reproductive organ and they play important role inthe early pregnant process. But there was no reports about whether theyexpress LeX and LeY or not. So firstly, we intend to study further thefunction how LeY influence the implantation of the mammal. Secondly, wewant to explore the function of LeX expressed on the early pregnant rats’endometriums. Thirdly, we intend to understand the expression andlocation of LeX and LeY in the oviduct and ovary of the early pregnantmouse then induce the possible functions. Method To examine the expression of LeX and LeY antigens inmouse endometriums ,oviduct and ovary of pregnancy after 1-7 day withimmunohistochemical method. To examine the expression of FUT9 whichis the key emzyme of LeX and the expression of FUT1and FUT4 whichform the LeY using the RT-PCR. To testify the expression of LeY in ovaryof the early pregnant mouse with Western blot. Result In mouse endometriums LeX expressed highly on the glandon the third day after pregnancy and it was relatively high on the third dayafter pregnancy and distributed on luminal epithelium. LeY expressedhighly on the fourth day after pregnancy, then it became to wane, but onthe luminal epithelium, it expressed lowly on the first day, during thefollowing day they maintained the moderate level. It expressed relativelyhigh on the implantation site. In oviduct LeX and LeY expressed lowly onthe first day after pregnancy, and on the third day after pregnancy, theyexpressed highly, then they become wane gradually. But on the seventh daythey still expressed at low level, and the positive sites were secretary cellsand ciliated cells. and LeY expressed strongly in the mouse ovary of thefourth day after pregnancy and mainly located in corpus luteum andstroma .The result of RT-PCR showed that the genes of FUT1and FUT4 aswell as FUT9 shared the same expressing tendency, and they all expressedhighly on the D3.The result of western-blot showed that LeY expressedstrongly on D4 in the mouse ovary Conclusion The study showed that LeX and LeY antigens expressedand located on the endometrium, oviduct and ovary of early pregnantmouse, the expression of genes of enzymes which regulated the formationof the two oligosaccharides was also observed. The result showed that LeXexpressed in a relatively high level in the pregnant mouse endometriumsduring the implantation window, Based on the known information onstructure and function, we suggest that it probably play an important role inthe implantation process.更多还原