牦牛繁殖周期不同阶段子宫中EGF、EGFR及HIF-2α的表达与定位研究  

Expression and Localization of EGF,EGFR and HIF-2α in the Uterus of Yak(Bos gruuniens)at Different Stages of Reproduction Cycle

作　　者：龙敏 崔燕[1] 王彪 庞鑫 余四九[1] 何俊峰[1] 白雪峰 王晓艳 LONG Min;CUI Yan;WANG Biao;PANG Xin;YU Si-Jiu;HE Jun-Feng;BAI Xue-Feng;WANG Xiao-Yan(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070

出　　处：《农业生物技术学报》2023年第8期1632-1640,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31972634)。

摘　　要：表皮生长因子(epidermal growth factor,EGF)及其受体(epithelial growth factor receptor,EGFR)可在一定程度上调节子宫内膜上皮细胞生长和存活,低氧诱导因子-2α(hypoxia inducible factor-2α,HIF-2α)是调节子宫内部环境及动物适应低氧环境的主要因子之一,三者均可影响子宫的发育及功能的正常发挥。本研究以不同时期牦牛(Bos grunniens)子宫为研究对象,采用免疫组织化学(immunohistochemistry,IHC)方法对EGF、EGFR及HIF-2α在牦牛子宫内的表达进行定位检测;利用qPCR和Western blot方法检测并分析EGF、EGFR以及HIF-2α在牦牛不同时期子宫中mRNA和蛋白水平的表达差异,探究其参与牦牛生殖发育的分子机制。IHC结果显示,EGF、EGFR和HIF-2α在3个时期子宫中均主要分布在牦牛子宫内膜上皮的胞质、子宫腺、血管内皮细胞、淋巴细胞;qPCR结果显示,EGF在黄体期的相对表达量显著高于卵泡期和妊娠期(P<0.05),EGFR和HIF-2α在卵泡期相对表达量显著高于黄体期和妊娠期(P<0.05);Western blot结果显示,EGF、EGFR和HIF-2α在妊娠期相对表达量显著高于卵泡期和黄体期(P<0.05)。综上所述,EGF、EGFR与HIF-2α在子宫中广泛分布,且三者在不同时期的表达存在差异,提示其可能在牦牛生殖过程中子宫的生理活动中发挥着重要作用。本研究进一步阐明EGF、EGFR和HIF-2α从发情到妊娠过程中在牦牛子宫中的表达变化,为研究其在低氧环境下牦牛繁殖周期不同阶段子宫内膜发育机制中的作用提供了基础资料。Epidermal growth factor(EGF)and its receptor(epithelial growth factor receptor,EGFR)can regulate the growth and survival of endometrial epithelial cells to a certain extent,hypoxia-inducible facto-2α(HIF-2α)is one of the main factors regulating the internal environment of the uterus and the adaptation of animals to the hypoxic environment,all of which can affect the development and normal function of the uterus.In this study,the expression of EGF,EGFR,and HIF-2αin the yak(Bos grunniens)uterus was localized and detected by immunohistochemistry(IHC),qPCR and Western blot.The expression of EGF,EGFR,and HIF-2α in the yak uterus at different stages of life was analyzed by qPCR and Western blot to investigate the molecular mechanisms involved in the reproductive development of yaks.IHC results showed that EGF,EGFR,and HIF-2α were mainly distributed in the cytoplasm,uterine gland,vascular endothelial cells,and lymphocytes of the yak endometrial epithelium in 3 periods of the uterus.The qPCR results showed that the relative expression of EGF in the luteal phase was significantly higher than that in the follicular phase and gestational stage(P<0.05),and the relative expression of EGFR and HIF-2α in the follicular phase was significantly higher than that in the luteal phase and pregnancy(P<0.05).The results of Western blot showed that the relative expression of EGF,EGFR,and HIF-2α was significantly higher than that in the follicular stage and luteal phase during pregnancy(P<0.05).In conclusion,EGF,EGFR,and HIF-2α were widely distributed in uterus,and the expression of the 3 were different at different times,suggested that they might play an important role in yak reproduction.This study further elucidates the changes of expression of EGF,EGFR,and HIF-2α in yak uterus from estrus to pregnancy,and provides basic information to study the role of the three factors in the development mechanism of endometrial of Yak at different stages of reproduction cycle.

关 键 词：牦牛 子宫 表皮生长因子(EGF) 表皮生长因子受体(EGFR) 低氧诱导因子-2α(HIF-2α) 

分 类 号：S857.22[农业科学—临床兽医学]