耐甲氧西林金黄色葡萄球菌诱导牛乳腺上皮细胞炎症反应的分子标志物研究

doi:10.11843/j.issn.0366-6964.2016.10.007

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (10): 1995-2002.doi: 10.11843/j.issn.0366-6964.2016.10.007

耐甲氧西林金黄色葡萄球菌诱导牛乳腺上皮细胞炎症反应的分子标志物研究

宋敏艳¹，韦艺媛^1，2，Muhammad Zahoor Khan¹，王新³，俞英^1*

（1.中国农业大学动物科技学院，农业部动物遗传育种与繁殖重点实验室&畜禽育种国家工程实验室，北京 100193；2.重庆市畜牧技术推广总站，重庆 401121； 3.西北农林科技大学食品学院，杨凌 712100）

收稿日期:2016-01-07 出版日期:2016-10-23 发布日期:2016-10-24
通讯作者: 俞英，博士生导师，主要从事动物健康性状改良及表观遗传调控机制研究，E-mail：yuying@cau.edu.cn
作者简介:宋敏艳（1986-），女，陕西咸阳人，博士生，主要从事奶牛表观遗传调控与抗病机理研究，E-mail：songminyanjiandan@163.com
基金资助:
国家自然科学基金(31272420)；国家奶牛产业技术体系专项资金（CARS-37-04B）；“十二五”国家科技支撑项目（2011BAD28B02）；教育部基本科研项目(2011JS006；Z109021306)；长江学者与创新团队发展计划（IRT1191）

Molecular Marker Study of Inflammatory Reaction in Bovine Mammary Epithelium Cell Line Induced by Methicillin-Resistant Staphylococcus aureus

SONG Min-yan¹，WEI Yi-yuan ^1，2，Muhammad Zahoor Khan¹，WANG Xin³，YU Ying^1*

(1.Key Laboratory of Animal Genetics，Breeding and Reproduction，Ministry of Agriculture & National Engineering Laboratory for Animal Breeding，College of Animal Science and Technology，China Agricultural University，Beijing 100193，China；2.Chongqing Animal Husbandry Techniques Extension Center，Chongqing 401121， China；3.College of Food Science and Engineering，Northwest A&F University，Yangling 712100， China)

Received:2016-01-07 Online:2016-10-23 Published:2016-10-24

摘要/Abstract

摘要：

旨在获得耐甲氧西林金黄色葡萄球菌（MRSA）诱导牛乳腺上皮细胞炎症反应的关键分子标志物。本研究以牛乳腺上皮细胞系Mac-T为试验材料，利用MRSA进行体外感染，通过与普通金葡菌感染组及未感染组进行比较，检测隐性乳房炎候选基因TRAPPC9、JAK2及炎症指示因子IL-6基因在MRSA感染不同时间点的基因表达量。结果显示，与未感染的对照组相比，MRSA 菌株体外感染Mac-T细胞6 h时，TRAPPC9基因表达量显著降低（P<0.05）；感染8 h时，JAK2基因表达量显著降低（P<0.05）；金葡菌菌株感染Mac-T细胞6 h时，TRAPPC9与JAK2基因表达量均显著降低（P<0.05）。与普通金葡菌相比，经MRSA处理6和10 h时，Mac-T细胞的TRAPPC9、JAK2及IL-6基因表达量均较高，且感染6 h时，MRSA处理组TRAPPC9基因表达量显著高于金葡菌处理组（P<0.05）。结果表明，MRSA及金葡菌感染Mac-T细胞后，3个基因表达变化规律基本相似，MRSA菌株感染更不易引起宿主细胞JAK2基因表达量的变化，而TRAPPC9基因则可作为MRSA金葡菌及普通金葡菌感染的首选分子标志物。

关键词: 耐甲氧西林金黄色葡萄球菌, 奶牛隐性乳房炎, 乳腺上皮细胞, TRAPPC9, JAK2

Abstract:

The aim of this study was to acquire key molecular marker related to inflammatory reaction of bovine mammary epithelium cells induced by Methicillin-resistant Staphylococcus aureus (MRSA).In this study，bovine mammary epithelium cell line (Mac-T) was infected with MRSA in vitro by compared with the cells treated by S.aureus as well the untreated cells.The expression levels of two candidate genes associated with subclinical mastitis ((trafficking protein particle complex 9(TRAPPC9)，Janus Kinase 2(JAK2)) and one pro-inflammatory gene (interleukin 6，IL-6) were detected and compared among infected groups and non-infected group at different time points.Comparing with the untreated cells，the results showed that the mRNA expression level of TRAPPC9 significantly decreased (P<0.05) in the Mac-T cells at 6 h post stimulation by MRSA，while that of JAK2 significantly decreased (P<0.05) at 8 h；And the expression levels of TRAPPC9 and JAK2 in the cells significantly decreased (P<0.05) at 6 h post infection by S.aureus.Compared with S.aureus stimuli，the expression levels of TRAPPC9，JAK2 and IL-6 in the Mac-T cells were moderately higher at 6 and 10 h post infection by MRSA.Especially，the expression level of TRAPPC9 was significantly higher in MRSA treated cells (P<0.05) compared to S.aureus treated cells at 6 h post infection.These results suggest that the genes expression changes in the Mac-T cells were similar after the infection of MRSA and S.aureus，while the gene expression changes of JAK2 in the cells was delayed from MRSA infection compared to S.aureus.TRAPPC9 can be used as the preferred molecular marker for detecting MRSA and S.aureus infection in bovine mammary epithelium cells.

Key words: Methicillin-resistant Staphylococcus aureus, bovine subclinical mastitis, mammary epithelium cell lines, TRAPPC9, JAK2

中图分类号:

S823
S813.3

宋敏艳，韦艺媛，Muhammad Zahoor Khan，王新，俞英. 耐甲氧西林金黄色葡萄球菌诱导牛乳腺上皮细胞炎症反应的分子标志物研究[J]. 畜牧兽医学报, 2016, 47(10): 1995-2002.

SONG Min-yan，WEI Yi-yuan，Muhammad Zahoor Khan，WANG Xin，YU Ying. Molecular Marker Study of Inflammatory Reaction in Bovine Mammary Epithelium Cell Line Induced by Methicillin-Resistant Staphylococcus aureus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(10): 1995-2002.

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耐甲氧西林金黄色葡萄球菌诱导牛乳腺上皮细胞炎症反应的分子标志物研究

Molecular Marker Study of Inflammatory Reaction in Bovine Mammary Epithelium Cell Line Induced by Methicillin-Resistant Staphylococcus aureus

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