一氧化氮合酶在奶牛临床型乳房炎乳腺组织中的表达、定位及功能分析  

Expression,Localization and Functional Analysis of NOS in Mammary Gland Tissues of Dairy Cows(Bos taurus)with Clinical Mastitis

作　　者：李建富 张博皓 陈文丽 袁宝 张勇[1,2,3] 赵兴绪 张全伟[1,2,3] LI Jian-Fu;ZHANG Bo-Hao;CHEN Wen-Li;YUAN Bao;ZHANG Yong;ZHAO Xing-Xu;ZHANG Quan-Wei(College of Life Science and Biotechnology,Gansu Agricultural University,Lanzhou 730070,China;College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学生命科学技术学院,兰州730070 [2]甘肃农业大学动物医学院,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》2025年第4期792-801,共10页Journal of Agricultural Biotechnology

基　　金：甘肃省高等学校产业支撑计划(2024CYZC-30);甘肃省创新驱动助力工程(GXH20240328-4);国家自然科学基金(U21A20262);甘肃省研究生“创新之星”(2023CXZX-635)。

摘　　要：一氧化氮合酶(nitric oxide synthase,NOS)是机体内源性一氧化氮(nitric oxide,NO)生成的关键酶,其催化生成的NO具有抗炎或促炎等多重作用。为了探究健康与患临床型乳房炎奶牛(Bos taurus)乳腺组织中NOS表达规律及其潜在功能,本研究以处于健康(control,Con)和临床型乳房炎(clinical mastitis,CM)状态的泌乳期荷斯坦奶牛乳腺组织为研究对象,利用苏木精-伊红(hematoxylin-eosin,HE)、免疫组织化学(immunohistochemistry,IHC)染色和实时荧光定量PCR(real-time fluorescence quantitative PCR,q RTPCR)分析奶牛乳腺组织中NOS分布及表达规律;通过生物信息学方法分析奶牛乳腺组织中NOS潜在作用。HE染色显示,CM组乳腺组织腺泡塌陷,伴随炎性细胞浸润;IHC染色显示,奶牛乳腺上皮细胞胞质中神经型一氧化氮合酶(neuronal NOS,nNOS)、诱导型一氧化氮合酶(inducible NOS,iNOS)和内皮型一氧化氮合酶(endothelial NOS,eNOS)呈阳性表达;qRT-PCR结果显示,与Con组相比,CM组奶牛乳腺组织nNOS和eNOS m RNA表达水平极显著下调(P<0.01),iNOS mRNA表达水平极显著上调(P<0.01);生物信息学分析显示,NOS可能通过调控NO与氨基酸合成代谢过程及离子转运过程参与CM发生。综上表明,NOS可能通过奶牛乳腺上皮细胞中NO生成调控乳腺炎症反应。本研究为深入揭示奶牛CM中NOS功能及调控机制提供参考。Nitric oxide synthase(NOS),a crucial enzyme responsible for production of endogenous nitric oxide(NO)in body,has multiple effects such as anti-inflammatory or pro-inflammatory.To investigate the expression patterns and potential functions of NOS in dairy cows(Bos taurus)with healthy or clinical mastitis(CM),the mammary gland tissues were collected from dairy cows with healthy(control,Con)and CM in this study.The distribution and expression patterns of NOS were in the mammary gland tissues of the Con and CM groups using hematoxylin-eosin(HE)staining,immunohistochemistry(IHC)and real-time fluorescence quantitative PCR(qRT-PCR).The potential role of NOS was predicted using different bioinformatics methods.HE displayed that the mammary acini were collapsed and couple with infiltration inflammatory cell in the CM group.IHC indicated that the positive expression of neuronal NOS(nNOS),inducible NOS(iNOS),and endothelial NOS(eNOS)were presented in the cytoplasm of mammary epithelial cells.Compared to the Con group,the n NOS and eNOS mRNA expression levels were extremely significantly downregulated(P<0.01),and iNOS m RNA expression levels was extremely significantly upregulated(P<0.01)in mammary gland tissues of the CM groups.Bioinformatics analysis revealed that NOS might participate in CM occurrence in regulation of NO and amino acid synthesis and metabolism,ion transport.In conclusion,the above results provide a reference for revealing the function and regulatory mechanism of NOS in cow CM.

关 键 词：临床型奶牛乳房炎 一氧化氮 一氧化氮合酶 生物信息学 功能分析 

分 类 号：S185[农业科学—农业基础科学] S823