奶牛子宫炎性腔液外泌体小RNA分布和组成分析  

Distribution and composition analysis of exosome-derived small RNA in inflammatory cavity fluid of cow uterus

作　　者：田凤[1] 常震宇 丁进 李成忠 冯妍 封紫嫣 倪和民[1] 盛熙晖[1] 齐晓龙[1] 邢凯 郭勇[1] 王相国[1] TIAN Feng;CHANG Zhenyu;DING Jin;LI Chengzhong;FENG Yan;FENG Ziyan;NI Hemin;SHENG Xihui;QI Xiaolong;XING Kai;GUO Yong;WANG Xiangguo

机构地区：[1]北京农学院动物科学技术学院,北京102206 [2]北京雄特牧业有限公司,北京100101 [3]昌平区动物卫生监督管理局十三陵防疫站,北京102200

出　　处：《北京农学院学报》2019年第4期95-99,共5页Journal of Beijing University of Agriculture

基　　金：国家重点研发计划项目（2017YFD0502200）;国家自然科学基金青年项目（31802263）;北京市教委科技计划一般项目（KM201810020007）;现代农业产业技术体系北京市奶牛创新团队（5075299027）

摘　　要：【目的】筛选奶牛子宫内膜炎诊断新型标志。【方法】采用贝博试剂盒分别从健康奶牛、患子宫内膜炎奶牛的子宫腔液中分离外泌体,透射电子显微镜检测外泌体形态和粒径大小,蛋白免疫印迹法检测外泌体蛋白标记CD9的表达,Trizol提取外泌体内RNA,Agilent 4200生物分析仪检测RNA的分布与组成。【结果】透射电子显微镜下奶牛子宫腔液外泌体呈囊泡形,有完整的膜结构,粒径大小为50~150nm之间;蛋白免疫印迹法检测到外泌体内均有CD9表达;生物分析仪检测显示子宫内膜炎症改变了子宫腔液内小RNA长度分布和组成。【结论】子宫腔液中小RNA变化可以作为奶牛子宫内膜炎症诊断的新型标志。【Objective】Screening new diagnostic markers of cow endometritis.【Methods】The exosomes was isolated from the uterine cavity of healthy cows and endometritis cows by Bebe kit.The morphology and size of exosomes were detected by transmission electron microscopy.The exosomes protein marker CD9 was detected by Western blotting.Exosomes RNA was extracted by Trizol,and RNA distribution and composition was analyzed by Agilent 4200 Bioanalyzer.【Results】Our results showed that the uterine cavity exosomes of the cows were vesicle-shaped under transmission electron microscope,that had a complete membrane structure with a particle size of about 50-150 nm.CD9 expression was detected in the exosomes.The bioanalyzer detection result showed that the distribution and composition of small RNA distribution in the uterine fluid were changed under endometrial inflammation condition.【Conclusion】Changes in exosome-derived small RNA expression and distribution from uterine cavity fluid can be used as new marker for the diagnosis of cow endometritis.

关 键 词：奶牛 子宫内膜炎 外泌体 小RNA 

分 类 号：S823.91[农业科学—畜牧学]