MVK和PMVK在临床型乳房炎奶牛乳腺中的表达定位及功能预测  

Expression,Localization and Functional Prediction of MVK and PMVK in Mammary Gland of Dairy Cows(Bos taurus)with Clinical Mastitis

作　　者：张博皓 张全伟[2,3] 蔺婷 戴丽君 白旭 赵兴绪 张勇[1,2,3] ZHANG Bo-Hao;ZHANG Quan-Wei;LIN Ting;DAI Li-Jun;BAI Xu;ZHAO Xing-Xu;ZHANG Yong(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;College of Life Science and Biotechnology,Gansu Agricultural University,Lanzhou 730070,China;Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070 [2]甘肃农业大学生命科学技术学院,兰州730070 [3]甘肃省动物生殖生理及繁殖调控重点实验室,兰州730070

出　　处：《农业生物技术学报》2023年第7期1430-1440,共11页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(U21A20262);甘肃省研究生“创新之星”项目(2023CXZX-635);宁夏回族自治区重点研发项目(2019BBF02027);甘肃省动物生殖生理及繁殖调控重点实验室(20JR10RA563)。

摘　　要：甲戊酸激酶(mevalonate kinase,MVK)和磷酸甲戊酸激酶(phosphomevalonate kinase,PMVK)与动物炎症发生和发展密切相关,而其在奶牛(Bos taurus)临床型乳房炎(clinical mastitis,CM)中的表达规律和作用机制尚不完全清楚。为探讨MVK和PMVK在正常与患CM奶牛乳腺组织中的表达规律及其潜在功能,本研究采集泌乳期正常(control,C)与患CM的荷斯坦奶牛乳腺组织(n=3/组),通过苏木精-伊红(hematoxylin-eosin,HE)染色观察奶牛乳腺组织病理变化,使用免疫组织化学(immunohistochemical,IHC)染色、免疫荧光(immunofluorescence,IF)染色和qRT-PCR分析奶牛乳腺组织中MVK和PMVK分布特征和表达规律;基于数据非依赖型模式(data-independent acquisition,DIA)蛋白质组学数据,通过生物信息学分析和探讨MVK和PMVK在奶牛CM中的作用。结果显示,MVK和PMVK主要定位于奶牛乳腺上皮细胞的细胞质;与C组相比,CM组中MVK和PMVK基因及其蛋白表达极显著下调(P<0.01);生物信息学结果提示,MVK与PMVK可能通过调控辅酶、氨基酸及脂肪酸合成代谢影响奶牛CM炎症发展。结果表明,MVK和PMVK参与调控奶牛CM发展,其表达下调可加剧乳腺炎症反应。本研究为奶牛CM中MVK和PMVK作用机制的研究提供参考。Mevalonate kinase(MVK)and phosphovalonate kinase(PMVK),the key enzymes of mevalonate pathway,are considered to be closely related to inflammatory response.However,the role and mechanism of MVK and PMVK in cows(Bos taurus)with clinical mastitis(CM)is unknown.The aim of this study was to explore the expression and distribution of MVK and PMVK,and further investigate their potential function in cows with CM.The mammary gland tissues of Holstein cows in Control(C,n=3)and CM group(n=3)were collected.The expression and distribution of MVK and PMVK in cow mammary gland tissues were analyzed by hematoxylin-eosin(HE)staining,immunohistochemical(IHC)staining,immunofluorescence(IF)staining and qRT-PCR;Meanwhile,based on the data-independent acquisition(DIA)proteomic data,the role of MVK and PMVK in CM was analyzed and discussed through bioinformatics.The results showed that MVK and PMVK were mainly localized in the cytoplasm of epithelial cells;Compared with the C group,the MVK and PMVK gene and its protein levels of in CM group were extremely significantly down regulated(P<0.01);The bioinformatics analysis showed that MVK and PMVK were involved in inflammatory regulation by affecting coenzyme metabolism,amino acid and fatty acid synthesis.In conclusion,the results indicated that MVK and PMVK were correlated with the occurrence and development of CM in cows,which provides important insights into the function and regulatory mechanism of MVK and PMVK in CM of cows.

关 键 词：临床型乳房炎 甲戊酸激酶(MVK) 磷酸甲戊酸激酶(PMVK) 数据非依赖型模式(DIA)蛋白质组学 功能分析 

分 类 号：S185[农业科学—农业基础科学] S823