与脐带间充质干细胞共培养促进奶牛乳腺上皮细胞乳蛋白合成及其机制  

Co-culture with umbilical cord mesenchymal stem cells promotes the synthesis and mechnism of milk protein in bovine mammary epithelial cells

作　　者：赵艳坤[1] 邵伟[1] 雒诚龙 武开乐 余雄[1] ZHAO Yankun SHAO Wei LUO Chenglong WU Kaile YU Xiong(College of Animal Science, Xinjiang Agricultural University, Xinjiang Meat Emulsions with Plant-eating Animal Nutrition Laboratory, Urumqi 830052, Chin)

机构地区：[1]新疆农业大学动物科学学院,新疆肉乳用草食动物营养实验室,新疆乌鲁木齐830052

出　　处：《细胞与分子免疫学杂志》2017年第2期185-189,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：奶产业体系资助(CARS-37);国家自然科学基金(31560645);新疆农业大学畜牧学博士后工作站资助;新疆维吾尔自治区高等学校科研计划项目(XJEDU2013S17);2013年度新疆研究生科研创新项目(xjau-2013-yjsky-XJGRI2013113)

摘　　要：目的研究脐带间充质干细胞(UCMSC)调节奶牛乳腺上皮细胞(BMEC)乳蛋白合成的可能作用机制。方法利用TranswellTM小室将UCMSC和BMEC进行双层共培养,BMEC单纯培养为对照组,胰岛素样生长因子1受体(IGF-1R)抑制剂AG1024处理细胞,ELISA检测上清IGF-1和β酪蛋白(CSN2)、κ酪蛋白(CSN3)含量变化,实时定量PCR检测Janus激酶2/信号转导子与转录激活子5(JAK2/STAT5)信号通路相关基因的相对表达丰度;再用JAK2信号通路阻断剂AG490孵育细胞,实时定量PCR检测CSN2、CSN3 mRNA的相对表达丰度。结果与UCMSC共培养后,BMEC的CSN2、CSN3合成量和CSN2、CSN3、JAK2、STAT5、E74样ETS转录因子5(ELF5)mRNA相对表达丰度均显著高于单纯培养的BMEC;AG1024处理后,显著降低BMEC的CSN2、CSN3合成量,显著降低CSN2、CSN3、JAK2、STAT5、ELF5 mRNA的相对表达丰度;给予AG490阻断后,显著降低CSN2、CSN3 mRNA相对表达丰度;在AG1024基础上,加入AG490后显著降低CSN2、CSN3 mRNA相对表达丰度。结论UCMSC能够通过IGF-1介导JAK2/STAT5信号通路,上调BMEC乳蛋白合成关键基因mRNA的表达丰度,促进其乳蛋白的合成。Objective To explore the possible mechanism of umbilical cord mesenchymal stem cels（ UCMSCs） regulating milk protein synthesis in bovine mammary gland epithelial cells（ BMECs）. Methods UCMSCs and BMECs were co-cultured by double-chamber TranswellTM,and other BMECs were cultured alone as a control group. Insulin like growth factor 1 receptor（ IGF-1R） inhibitor AG1024 was used to treat cells. IGF-1,β casein（ CSN2） and κ casein（ CSN3） content in the supernatants were determined by ELISA; the relative expression abundance of Janus kinase and signal transducer and activator of transcription factor（ JAK / STAT） signaling pathway-related genes were detected by quantitative real-time PCR（ qRT-PCR）; and after the cells were treated with JAK2 signal blocker AG490,qRT-PCR was performed to test the relative expression abundance of CSN2 and CSN3 mRNAs. Results CSN2, CSN3 synthetic quantity, and the relative expression abundance of CSN2,CSN3,JAK2,STAT5,E74-like ETS transcription factor 5（ ELF5） mRNAs of the co-culture group were significantly higher than those of the control group. After treated with AG1024,the co-culture group showed remarkably decreased CSN2,CSN3 synthesis in BMECs as wel as the decreased relative expression abundance of CSN2,CSN3,JAK2,STAT5,ELF5 mRNAs. After blocked with AG490,the relative expression abundance of CSN2 and CSN3 mRNAs were reduced significantly in the co-culture group. In addition,the expressions of CSN2 and CSN3 mRNAs were inhibited significantly when both AG1024 and AG490 were supplemented. Conclusion UCMSCs can mediate JAK2 / STAT5 signaling pathwayvia IGF-1,and increase the expressions of milk protein synthesis key genes in BMECs,thus promoteing the synthesis of milk protein.

关 键 词：脐带间充质干细胞(UCMSC) 牛乳腺上皮细胞(BMEC) 共培养 乳蛋白 β酪蛋白(CSN2) κ酪蛋白(CSN3) 

分 类 号：S823.91[农业科学—畜牧学] R446.61[农业科学—畜牧兽医]