冷诱导RNA结合蛋白参与小鼠H1N1甲型流感病毒感染的应答

doi:doi: 10.11843/j.issn.0366-6964.2016.08.016

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (8): 1652-1657.doi: doi: 10.11843/j.issn.0366-6964.2016.08.016

冷诱导RNA结合蛋白参与小鼠H1N1甲型流感病毒感染的应答

聂培婷^1，2，汤承¹，岳华^1*

（1.西南民族大学生命科学与技术学院，成都 610041；2.新疆天康畜牧生物技术股份有限公司制药事业部，乌鲁木齐 830011）

收稿日期:2015-12-28 出版日期:2016-08-23 发布日期:2016-09-08
通讯作者: 岳华，教授，E-mail：yhua900@163.com
作者简介:聂培婷（1989-），女，新疆霍城人，主要从事感染与免疫学研究，E-mail：540016852@qq.com
基金资助:
国家自然科学基金（31172307）；四川省教育厅创新团队（13TD0057）

Cold-inducible RNA Binding Protein of Mice Involved in Response to Influenza A Virus Infection in Mice

NIE Pei-ting ^1，2，TANG Cheng ¹，YUE Hua1^*

（1.College of Life Science and Technology，Southwest University for Nationalities，Chengdu 610041，China；
2.Xinjiang Tecon Animal Husbandry Bio-Technology CO.，LT，Urumqi 830011，China）

Received:2015-12-28 Online:2016-08-23 Published:2016-09-08

摘要/Abstract

摘要：

冷诱导RNA结合蛋白（CIRP）是哺乳动物间高度保守的多功能蛋白，但其在病毒感染中的作用尚不清楚，本研究旨在探讨CIRP对甲型流感病毒感染的应答。用H1N1甲型流感病毒PR-8株滴鼻接种成年BALB/c小鼠，分别于感染后24、48、72、96、120 h随机处死3只，用荧光定量RT-PCR和免疫组化法分别检测小鼠心、肝、脾和肺中CIRP基因和蛋白质的表达水平。基因定量检测结果显示，感染小鼠各被检器官中Cirp mRNA的转录量显著升高；免疫组化试验结果显示，CIRP在感染小鼠心肌细胞、肺泡上皮细胞、肺支气管上皮细胞和脾淋巴细胞的细胞质中表达量均有不同程度的升高。综上可见，CIRP参与机体对H1N1甲型流感病毒感染的应答，由于CIRP对炎性因子产生过程具有显著的调节作用，其在流感病毒感染过程中的作用值得深入研究。

关键词: , 冷诱导RNA结合蛋白, 甲型流感病毒, 荧光定量RT-PCR, 免疫组化

Abstract:

Cold induced RNA binding protein (CIRP) is a highly conserved multifunctional protein among vertebrates，but its role in viral infection is unclear.The objective of this paper was to explore the response of CIRP to influenza virus infection.The BALB/c mice were intranasal inoculated with H1N1 influenza A virus.Three mice were randomly selected in each group and euthanized at 24，48，72，96，120 h post infection.The heart，liver，spleen and lung of each mouse were collected.Real-time RT-PCR and immunohistochemistry (IMH) were used to detect the transcription of Cirp mRNA and expression of CIRP in each organ.The results of Real-time RT-PCR exhibited that the Cirp mRNA significantly increased in all detected organs.The results of IMH test showed that CIRP protein was highly expressed in cytoplasm of myocardial cells，alveolar epithelial cells，bronchial epithelium cells and spleen lymphocytes.In summary，CIRP is involved in the response to influenza A virus infection，because CIRP can significantly regulate the production of inflammatory factors，the role of CIRP in influenza virus infection should be further studied.

Key words: cold-inducible RNA binding protein, influenza A virus, real-time RT-PCR, immunohistochemistry

中图分类号:

S852.23

聂培婷，汤承，岳华. 冷诱导RNA结合蛋白参与小鼠H1N1甲型流感病毒感染的应答[J]. 畜牧兽医学报, 2016, 47(8): 1652-1657.

NIE Pei-ting，TANG Cheng，YUE Hua. Cold-inducible RNA Binding Protein of Mice Involved in Response to Influenza A Virus Infection in Mice[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(8): 1652-1657.

参考文献

［1］BURD C G，DREYFUSS G.Conserved structures and diversity of functions of RNA-binding proteins［J］. Science，1994，265(5172)：615-621.
［2］NISHIYAMA H，ITOH K，KANEKO Y，et al.A glycine-rich RNA-binding protein mediating cold-inducible suppression of mammalian cell growth［J］.J Cell Biol，1997，137(4)：899-908.
［3］XUE J H，NONOGUCHI K，FUKUMOTO M，et al.Effects of isehemia and H2O2：on the cold stress protein CIRP expression in rat neuronal cells［J］.Free Radic Biol Med，1999，27(11-12)：1238-1244.
［4］WELLMANN S，BÜHRER C，MODEREGGER E，et al.Oxygen-regulated expression of the RNA-binding proteins RBM3 and CIRP by a HIF-l-independent mechanism［J］.J Cell Sci，2004，117(Pt 9)：1785-1794.
［5］PAN F，ZARATE J，CHOUDHURY A，et al.Osmotic stress of salmon stimulates upregulation of a cold inducible RNA binding protein (CIRP) similar to that of mammals and amphibians［J］.Biochimie，2004，86(7)：451-461.
［6］YANG C，CARRIER F.The UV-inducible RNA-binding protein A18 (A18 hnRNP) plays a protective role in the genotoxic stress response［J］.J Biol Chem，2001，276(50)：47277-47284.
［7］DE LEEUW F，ZHANG T，WAUQUIER C，et al.The cold-inducible RNA-binding protein migrates from the nucleus to cytoplasmic stress granules by a methylation-dependent mechanism and acts as a translational repressor［J］.Exp Cell Res，2007，313(20)：4130-4144.
［8］BROCHU C，CABRITA M A，MELANSON B D，et al.NF-κb-dependent role for cold-inducible RNA binding protein in regulating interleukin 1β［J］.PLoS One，2013，8(2)：e57426.
［9］QIANG X，YANG W L，WU R，et al.Cold-inducible RNA-binding protein (CIRP) triggers inflammatory responses in hemorrhagic shock and sepsis［J］.Nat Med，2013，19(11)：1489-1495.
［10］ZHOU M，YANG W L，JI Y，et al.Cold-inducible RNA-binding protein mediates neuroinflammation in cerebral ischemia［J］. Biochim Biophys Acta，2014，1840(7)：2253-2261.
［11］NEUMANN G，NODA T，KAWAOKA Y.Emergence and pandemic potential of swine-origin H1N1 influenza virus［J］.Nature，2009，459(7249)：931-939.
［12］TEIJARO J R，WALSH K B，RICE S，et al.Mapping the innate signaling cascade essential for cytokine storm during influenza virus infection［J］. Proc Natl Acad Sci U S A，2014，111(10)：3799-3804.
［13］LA GRUTA N L，KEDZIERSKA K，STAMBAS J，et al.A question of self-preservation：immunopathology in influenza virus infection［J］. Immunol Cell Biol，2007，85(2)：85-92.
［14］聂培婷，汤承，岳华.H1N1甲型流感病毒在BHK21中的增殖规律［J］.西南民族大学学报•自然科学版，2012，38(5)：764-769.
NIE P T，TANG C，YUE H.Study on the proliferation profile of influenza A virus in BHK21 cells［J］.Journal of Southwest University for Nationalities•Natural Science Edition，2012，38(5)：764-769.(in Chinese)
［15］周鸿淼，汤承，岳华，等.冷诱导RNA结合蛋白参与小鼠对LPS的应答［J］.畜牧兽医学报，2014，45(8)：1348-1354.
ZHOU H M，TANG C，YUE H，et al.Cold-inducible RNA binding protein involves in response to LPS in mice［J］.Acta Veterinaria et Zootechnica Sinica，2014，45(8)：1348-1354.(in Chinese)
［16］吴巧，张斌，汤承，等.H5N1禽流感病毒感染小鼠后内参基因的筛选［J］.中国畜牧兽医，2013，40(9)：55-60.
WU Q，ZHANG B，TANG C，et al.Selection of reference gene in mice infected with H5N1 avian influenza virus［J］.China Animal Husbandry & Veterinary Medicine，2013，40(9)：55-60.(in Chinese)
［17］LIVAK K J，SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-△△Ct method［J］.Methods,2001，25(4)：402-408.
［18］蔡文琴，王伯沄.实用免疫细胞化学与核酸分子杂交技术［M］.成都：四川科学技术出版社，1994.
CAI W Q，WANG B Y.Practical immunocytochemistry and nucleic acid hybridization techniques［M］.Chengdu：Sichuan Science and Technology Press，1994.(in Chinese)
［19］SAITO K，FUKUDA N，MATSUMOTO T，et al.Moderate low temperature preserves the stemness of neural stem cells and suppresses apoptosis of the cells via activation of the cold-inducible RNA binding protein［J］.Brain Res， 2010，1358：20-29.
［20］SHAPIRA S D，GAT-VIKS I，SHUM B O，et al.A physical and regulatory map of host-influenza interactions reveals pathways in H1N1 infections［J］.Cell，2009，139(7)：1255-1267.
［21］PREDICALA R，ZHOU Y.The role of Ran-binding protein 3 during influenza A virus replication［J］. J Gen Virol，2013，94(Pt 5)：977-984.
［22］PLANZ O.Development of cellular signaling pathway inhibitors as new antivirals against influenza［J］.Antiviral Res，2013，98(3)：457-468.
［23］LUDWIG S，PLANZ O.Influenza viruses and the NF-κB signaling pathway—towards a novel concept of antiviral therapy［J］.Biol Chem，2008，389(10)：1307-1312.
［24］CANNON G，CALLAHAN M A，GRONEMUS J Q，et al.Early activation of MAP kinases by influenza A virus X-31 in murine macrophage cell lines［J］.PLoS One，2014，9(8)：e105385.
［25］NIMMERJAHN F，DUDZIAK D，DIRMEIER U，et al.Active NF- kappaB signalling is a prerequisite for influenza virus infection［J］.J Gen Virol，2004，85(Pt 8)：2347-2356.

[1]	. 征订目录[J]. 畜牧兽医学报, 2019, 50(9): 0-0.
[2]	郑兰兰, 朱静静, 王盼, 舒燕, 梁青青, 李炳晓, 王超群, 魏战勇. 猪δ冠状病毒TaqMan荧光定量RT-PCR检测方法的建立与初步应用[J]. 畜牧兽医学报, 2019, 50(6): 1261-1267.
[3]	郑兰兰, 朱静静, 王盼, 舒燕, 梁青青, 李炳晓, 王超群, 魏战勇. 猪δ冠状病毒TaqMan荧光定量RT-PCR检测方法的建立与初步应用[J]. 畜牧兽医学报, 2019, 50(6): 1261-1267.
[4]	郭紫晶, 何琪富, 汤承, 张斌, 岳华. 检测纽布病毒的Real-time RT-PCR方法的建立与应用[J]. 畜牧兽医学报, 2019, 50(4): 893-900.
[5]	郭紫晶, 何琪富, 汤承, 张斌, 岳华. 检测纽布病毒的Real-time RT-PCR方法的建立与应用[J]. 畜牧兽医学报, 2019, 50(4): 893-900.
[6]	段世豪, 路建彪, 李旭勇, 刘成, 张树金, 殷国政, 司振书, 李玉保. 鸡马立克病肝组织肿瘤病变与热休克蛋白转录、表达的相关性研究[J]. 畜牧兽医学报, 2019, 50(2): 415-421.
[7]	段世豪, 路建彪, 李旭勇, 刘成, 张树金, 殷国政, 司振书, 李玉保. 鸡马立克病肝组织肿瘤病变与热休克蛋白转录、表达的相关性研究[J]. 畜牧兽医学报, 2019, 50(2): 415-421.
[8]	禇颖, 马文杰, 胡卫国, 陈茹, 邱建华, 郭慧君, 李宏梅. 检测鸡TRIM25蛋白的免疫组化方法[J]. 畜牧兽医学报, 2018, 49(4): 865-870.
[9]	禇颖, 马文杰, 胡卫国, 陈茹, 邱建华, 郭慧君, 李宏梅. 检测鸡TRIM25蛋白的免疫组化方法[J]. 畜牧兽医学报, 2018, 49(4): 865-870.
[10]	黄莉,谢芝勋，蓝元喜，谢丽基，邓显文，范晴，罗思思，谢志勤，黄娇玲，张艳芳，王盛，曾婷婷. 禽呼肠孤病毒感染对SPF鸡外周血淋巴细胞中天然免疫相关基因转录的影响[J]. 畜牧兽医学报, 2017, 48(1): 116-123.
[11]	黄莉,谢芝勋，蓝元喜，谢丽基，邓显文，范晴，罗思思，谢志勤，黄娇玲，张艳芳，王盛，曾婷婷. 禽呼肠孤病毒感染对SPF鸡外周血淋巴细胞中天然免疫相关基因转录的影响[J]. 畜牧兽医学报, 2017, 48(1): 116-123.
[12]	张琦，黄娇娇，杨彩侠，王宇祥，张慧，赵建国，李辉. CRISPR/Cas9介导RB1基因敲除及其在鸡前脂肪细胞分化、增殖中的功能研究[J]. 畜牧兽医学报, 2016, 47(9): 1775-1784.
[13]	王苗，罗军，许会芬，朱江江，赫秋亚，姚大为，史怀平. 山羊INSIG1基因超表达对乳腺上皮细胞中脂质合成的影响[J]. 畜牧兽医学报, 2016, 47(9): 1806-1816.
[14]	周作勇，邵骜骏，徐婷婷，陈家磊，胡世君. 基于高通量测序的猪鞭虫雌、雄虫差异基因表达分析[J]. 畜牧兽医学报, 2016, 47(9): 1914-1923.
[15]	唐丽，辛丹宁，田春洋，崔恒敏. 钒对雏鸡大脑皮层γ-氨基丁酸表达的影响[J]. 畜牧兽医学报, 2016, 47(9): 1940-1946.

冷诱导RNA结合蛋白参与小鼠H1N1甲型流感病毒感染的应答

Cold-inducible RNA Binding Protein of Mice Involved in Response to Influenza A Virus Infection in Mice

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价