中国畜牧兽医2025，Vol.52Issue(6)：2691-2704,14.DOI:10.16431/j.cnki.1671-7236.2025.06.022
夏南牛子宫外泌体miRNA在牛胚胎发育和着床中的作用研究
Study on the Role of miRNA from Uterine Exosomes in Embryo Development and Implantation of Xianan Cattle
摘要
Abstract
[Objective]This study was aimed to explore the expression differences of miRNA in uterine flushing exosomes of cattle with different embryo numbers obtained after superovulation,so as to clarify the regulatory mechanism of miRNA in uterine flushing exosomes during embryo development and implantation in cattle.[Method]Fifteen Xianan cattle aged 3-6 years with 480-600 kg were selected for superovulation,synchronized estrus and artificial insemination.On the 7th day after artificial insemination,the uterus in Xianan cattle was washed to obtain blastocysts.According to the international average embryo production level of 5.5 embryos as the standard,three Xianan cattle(experimental group)with the highest number of blastocysts obtained and two Xianan cattle(control group)with the lowest number of blastocysts obtained were selected.Uterine flush embryo fluid was collected for isolation and identification of exosomes,and exosomal miRNA was extracted for sequencing analysis.Target genes of differentially expressed miRNA were predicted,and GO function and KEGG pathway enrichment analysis on the target genes were performed.Four miRNAs(bta-miR-2888,bta-miR-11987,bta-miR-2484 and bta-miR-27a-3p)were randomly selected for Real-time quantitative PCR validation.[Result]The vesicle diameters of 5 samples was 30-150 nm,which was consistent with the characteristics of exosomes.A total of 14 differentially expressed miRNAs were screened,with 9 miRNAs upregulated and 5 miRNAs downregulated.A total of 8 703 target genes were predicted from differently expressed miRNAs.GO function enrichment analysis indicated that the target genes were primarily enriched in terms related to functions such as the regulation of developmental process,regulation of multicellular organismal development,regulation of transport,positive regulation of developmental process,cell junction,and others.KEGG pathway enrichment analysis indicated that the significantly enriched signaling pathways such as MAPK and Hedgehog,which were related to embryonic development and implantation,suggesting that exosomes in uterine flush embryo fluid might be involved in the regulation of embryonic development and implantation.Real-time quantification PCR results of 4 miRNAs were consistent with RNA-Seq results.[Conclusion]This study isolated exosomes from the uterine fluid flush in Xianan cattle,enriching the species investigated in exosome research,and providing valuable experience for future exosome research in Xianan cattle.There were 14 differentially expressed miRNAs were identified in this study,of which 8 miRNAs were associated with embryonic development and implantation.The results suggested that the change of exosome miRNA in uterus might influence embryonic development and implantation in cattle.关键词
夏南牛/外泌体/miRNA/胚胎发育/胚胎着床Key words
Xianan cattle/exosome/miRNA/embryo development/embryo implantation分类
农业科技引用本文复制引用
张志浩,施巧婷,王二耀,鲁立刚,张子敬,王香南,闵佳,韩艺伟,彭晟坤,栾曼茹,刘奥兵..夏南牛子宫外泌体miRNA在牛胚胎发育和着床中的作用研究[J].中国畜牧兽医,2025,52(6):2691-2704,14.基金项目
河南省肉牛产业技术体系项目(HARS-22-13-S) （HARS-22-13-S）
国家肉牛牦牛产业技术体系(CARS-37) （CARS-37）
河南省重大科技专项"地方畜禽精准快育种体系构建与新品种选育"(221100110200) （221100110200）
河南省农业科学院科技创新团队项目"牛羊繁育与生产"(2024TD25) （2024TD25）
