PMSG联合GnRH对小鼠胚胎着床的影响  

Effect of PMSG Combined with GnRH on Embryo Implantation in Mice(Mus musculus)

作　　者：李桂琳 袁文 杨静怡 李相运[1] 吴兴龙[1] LI Gui-Lin;YUAN Wen;YANG Jing-Yi;LI Xiang-Yun;WU Xing-Long(College of Animal Science and Technology,Hebei Agricultural University,Baoding 071000,China)

机构地区：[1]河北农业大学动物科技学院,保定071000

出　　处：《农业生物技术学报》2023年第5期1013-1021,共9页Journal of Agricultural Biotechnology

基　　金：河北农业大学引进人才科研专项(YJ201952)。

摘　　要：孕马血清促性腺激素(pregnant mare serum gonadotropin,PMSG)联合促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)通过刺激小鼠(Mus musculus)卵巢可提高繁殖效率,但是其对胚胎着床的影响尚不明确。本研究旨在探讨PMSG联合GnRH方案对小鼠胚胎着床的影响。实验组腹腔注射7.5 IU PMSG,48 h后再注射3.5μg GnRH类似物亮丙瑞林(简称P+G处理组),对照组注射生理盐水;将正常囊胚移植到假孕小鼠子宫中,5 d后比较不同处理组胚胎移植后的着床率,检测着床期子宫内膜容受性标记分子—白血病抑制因子(leukemia inhibitory factor,Lif)的基因表达;采用转录组数据分析P+G处理对子宫内膜容受性的影响,利用qRT-PCR和Western blot验证测序结果的准确性。结果显示,P+G假孕受体的胚胎着床率显著低于对照组(P<0.05),并且P+G处理的子宫内膜容受性标记基因Lif表达显著降低(P<0.05)。转录组测序分析发现751个差异表达基因,其中587个上调表达、164个下调表达;GO和KEGG分析显示,P+G处理后差异表达基因主要富集在免疫反应、调节免疫系统过程、细胞黏附分子通路以及细胞因子间的相互作用通路;qRT-PCR和Western blot验证结果显示,P+G处理使Lif基因及蛋白表达均下调,Lcn2和Prap1基因及蛋白表达均上调,变化趋势与转录组测序结果一致。本研究为深入探讨PMSG和GnRH参与胚胎着床的调控机制提供一定的理论基础。Pregnant mare serum gonadotropin(PMSG)combined with gonadotropin-releasing hormone(GnRH)can improve reproductive efficiency by stimulating the ovaries of mice(Mus musculus),but its effect on embryo implantation remains unclear.This study aimed to explore the effect of PMSG combined with GnRH on embryo implantation in mice.In the experiment group,mice were used to inject 3.5μg of GnRH agonist leuprorelin 48 h after 7.5 IU of PMSG injection(referred to as P+G group),and the control group was injected twice with saline.Normal blastocysts were transferred into the uterus of pseudo-pregnant mice in the P+G and control groups,then embryo implantation rates were analyzed after 5 d embryo transfer,and the expression of endometrial receptivity marker gene-leukemia inhibitory factor(Lif)was detected.Transcriptome sequencing data were analyzed to reveal the effect of P+G on endometrial receptivity,and qRTOnlinePCR and Western blot were used to verify the data accuracy.The results showed that the embryo implantation rate of P+G group was significantly lower than that of the control group(P<0.05),and the expression of endometrial receptivity marker gene Lif was significantly reduced(P<0.05)in the P+G group.Transcription group sequencing analysis found 751 differentially expressed genes(DEGs),of which 587 up-regulated and 164 down-regulated.GO and KEGG analysis found that DEGs mainly enriched in the immune response,immune system process,cell adhesion molecules(CAMs),and cytokine-cytokine receptor interaction pathways.The results of qRT-PCR and Western blot showed that the expression of Lif gene and protein were down-regulated and the expression of Lcn2 and Prap1 gene and protein were up-regulated in the P+G group,which was consistent with the results of transcriptome sequencing.This study provides a certain theoretical basis for indepth exploration of the regulatory mechanism of PMSG and GnRH involved in embryo implantation.

关 键 词：孕马血清促性腺激素(PMSG) 促性腺激素释放激素(GnRH) 子宫内膜容受性 着床率 胚胎移植 

分 类 号：S811.4[农业科学—畜牧学]