TLRs介导的信号通路在马链球菌感染RAW264.7细胞中的作用

doi:10.16431/j.cnki.1671-7236.2016.05.023

›› 2016, Vol. 43 ›› Issue (5): 1270-1276.doi: 10.16431/j.cnki.1671-7236.2016.05.023

TLRs介导的信号通路在马链球菌感染RAW264.7细胞中的作用

杜润慈¹, 刘建华¹, 史茜², 付强¹, 冉多良¹

1. 新疆农业大学动物医学学院, 乌鲁木齐 830052;
2. 新疆出入境检验检疫局动植物检疫中心, 乌鲁木齐 830063

收稿日期:2015-10-26 出版日期:2016-05-20 发布日期:2016-05-23
通讯作者: 冉多良 E-mail:xjrdl7@163.com
作者简介:杜润慈(1991-),女,新疆乌鲁木齐人,硕士,研究方向:动物传染病诊断与防治,E-mail:13350566383@163.com
基金资助:
国家科技支撑计划项目(2012BAD46B00)

Effect of TLRs Mediated Signal Pathway on Streptococcus equi Infection in RAW264.7 Cells

DU Run-ci¹, LIU Jian-hua¹, SHI Qian², FU Qiang¹, RAN Duo-liang¹

1. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China;
2. Xinjiang Exit-import Inspection and Quarantine Bureau, Urumqi 830063, China

Received:2015-10-26 Online:2016-05-20 Published:2016-05-23

摘要/Abstract

摘要： 为探究Toll样受体(TLRs)介导的信号通路在马链球菌马亚种(S.equi)感染小鼠巨噬细胞RAW264.7中的作用,收集S.equi感染后不同时间点的RAW264.7细胞,提取总RNA并反转录成cDNA,利用实时荧光定量PCR技术检测细胞Toll样受体1、2、6(TLR1、TLR2、TLR6)、接头蛋白骨髓分化蛋白88(MyD88)及细胞因子IL-1、IL-6、IL-10、IL-12、TNF-αmRNA的表达情况。结果显示,S.equi感染RAW264.7细胞后6h时,TLR1、TLR2、TLR6与MyD88mRNA水平均较对照组没有显著差异(P>0.05);感染后12h时,TLR1、TLR2和TLR6mRNA表达量未出现明显上升(P>0.05),而MyD88mRNA水平极显著升高(P<0.01);感染后24h时,TLR1、TLR2和TLR6mRNA表达水平出现极显著升高(P<0.01),MyD88mRNA表达没有显著变化(P>0.05),且IL-10和IL-12mRNA水平与对照组相比极显著升高(P<0.01),IL-1、IL-6和TNF-αmRNA水平均极显著下降(P<0.01)。结果表明,TLRs介导的信号通路参与S.equi感染RAW264.7细胞的免疫应答反应。

关键词: 马链球菌马亚种; RAW264.7细胞; Toll样受体; 骨髓分化蛋白88; 细胞因子

Abstract: To investigate the effects of Toll-like receptors (TLRs) mediated signaling pathway in mouse macrophage RAW264.7 cells on Streptococcus equi subspecies equi (S.equi) infection, RAW264.7 cells were infected with S.equi for different time intervals.The mRNA expression levels of TLR1, TLR2, TLR6, myeloid differentiation factor 88 (MyD88), IL-1, IL-6, IL-10, IL-12 and TNF-α were detected by fluorescence quantitative Real-time PCR.The results showed that at 6 h post-infection, the expression levels of TLR1, TLR2, TLR6 and MyD88 in S.equi infected RAW264.7 cells had no significant difference compared with control group (P>0.05).At 12 h post-infection, the mRNA expression levels of TLR1, TLR2 and TLR6 had no obviously change(P>0.05), while MyD88 mRNA expression level was extremely significantly increased (P<0.01).At 24 h post-infection, the mRNA levels of TLR1, TLR2, TLR6 were extremely significantly increased (P<0.01), the mRNA level of MyD88 had no significant difference (P>0.05), meanwhile, compared with uninfected cells, IL-10 and IL-12 mRNA expression were extremely significant increased in S.equi infected RAW264.7 cells (P<0.01), but IL-1, IL-6 and TNF-α mRNA levels were extremely significantly decreased (P<0.01).These results indicated that the signaling pathway mediated by TLRs participated in the immune response against S.equi.

Key words: Streptococcus equi subspecies equi; RAW264.7 cells; Toll-like receptor; MyD88; cytokine

中图分类号:

Q2-33

杜润慈, 刘建华, 史茜, 付强, 冉多良. TLRs介导的信号通路在马链球菌感染RAW264.7细胞中的作用[J]. , 2016, 43(5): 1270-1276.

DU Run-ci, LIU Jian-hua, SHI Qian, FU Qiang, RAN Duo-liang. Effect of TLRs Mediated Signal Pathway on Streptococcus equi Infection in RAW264.7 Cells[J]. , 2016, 43(5): 1270-1276.

参考文献

[1] 郭纪珂,王晓钧.马腺疫[J].畜牧兽医科技信息,2013,10:16-18.
[2] Flock M,Jacobsson K,Frykberg L,et al.Recombinant Streptococcus equi proteins protect mice in challenge experiments and induce immune response in horses[J].Infect Immun,2004,72(6):3228-3236.
[3] Flock M,Karlstrom A,Lannerqard J,et al.Protective effect of vaccination with recombinant proteins from Streptococcus equi subspecies equi in a strangles model in the mouse[J].Vaccine,2006,24(19):4144-4151.
[4] 徐全圆.马腺疫链球菌的分离鉴定及FN结合蛋白免疫原性的研究[D].乌鲁木齐:新疆农业大学,2014.
[5] 马思慧,杨欢,吴天成,等.Toll样受体信号传导通路的研究进展[J].中国畜牧兽医,2014,8(41):160-164.
[6] 郎需龙,冉会玲,王秀然,等.猪链球菌2型分解代谢控制蛋白A经NF-κB/MAPK通路对巨噬细胞RAW264.7影响的研究[J].中国畜牧兽医,2013,7(40):21-24.
[7] Miller L S.Toll-like receptors in skin[J].Adv Dermatol,2008,24(24):71-87.
[8] Takeda K,Kaisho T.Toll-like receptors[J].Annu Rev Immunol,2003,21(1):335-376.
[9] Takeuchi O,Hoshino K,Kawai T,et al.Differential roles of TLR2 and TLR4 in recognition of gram-negative and gram-positive cell wall components[J].Immunity,1999,11(4):443-451.
[10] Echchannaoui H,Frei K,Schnell C,et al.Toll-like receptor 2-deficient mice are highly susceptible to Streptococcus pneumoniae meningitis because of reduced bacterial clearing and enhanced inflammation[J].J Infect Dis,2002,186(6):798-806.
[11] Takeuchi O,Hoshino K,Akira S.Cutting edge:TLR2-deficient and MyD88 deficient mice are highly susceptible to Staphylococcus aureus infection[J].J Immunity,2000,165(10):5392-5396.
[12] Florindo H F,Pandit S,Goncalves L M D,et al.Streptococcus equi antigens adsorbed onto surface modified poly-ε-caprolactone microspheres induce humoral and cellular specific immune responses[J].Vaccine,2008,26(33):4168-4177.
[13] Parkinson N J,Robin C,Newton J R,et al.Molecular epidemiology of strangles outbreaks in the UK during 2010[J].Vet Rec,2011,168(25):666.
[14] Triantafilou M,Gamper F G.Membrane sorting of toll-like receptor(TLR)-2/6 and TLR2/1 heterodimers at the cell surface determines heterotypic associations with CD36 and intracellular targeting[J].Biol Chem,2006,281(41):31002-31011.
[15] Alexopoulou L,Holt A C,Medzhitov R,et al.Recognition of double-stranded RNA and activation of NF-kappa B by Toll-like receptor 3[J].Nature,2001,413(6875):732-738.
[16] Palsson-McDermott E M,O'Neill L A.Signal transduction by the lipoplysaccharide receptor,Toll-like receptor-4[J].Immunology,2004,62(2):113-153.
[17] Hayashi F,Smith K D,Ozinsky A,et al.The innate immune response to bacterial flagellin is mediated by toll-like receptor 5[J].Nature,2001,410(6832):1099-1103.
[18] Sandor F,Buc M.Toll-like receptors Ⅱ distribution and pathways involved in TLR signaling[J].Folia Biol(Praha),2005,51(6):188-197.
[19] Barton G M,Medzhitov R.Toll-like receptor signaling pathways[J].Science,2003,300(5625):1524-1525.
[20] Lecours M P, Segura M, Claude L, et al. Characterization of porcine dendritic cell response to Streptococcus suis[J].Veterinary Research,2011,42(42):1-12.
[21] Takeuchi O,Kaufmann A,Grote K,et al.Cutting edge:Preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-2 activates immune cells through a Toll-like receptor 2 and MyD88 dependent signaling pathway[J].J Immunol,2000,164(2):554-557.
[22] Takeuchi O,Kawai T,Muhlradt P F,et al.Discrimination of bacterial lipoproteins by Toll-like receptor 6[J].Int Immunol,2001,13(7):933.
[23] Rönnberg E,Guss B,Pejler G.Infection of mast cells with live streptococci causes a Toll-like receptor 2 and cell-cell contact-dependent cytokine and chemokine response[J].Infect Immun,2010,78(2):854-864.
[24] 张晓燕,蒋小武,杨岭.HEp-2细胞感染猪链球菌2型后细胞因子mRNA表达变化[J].中国预防兽医学报,2014,36(9):696-700.

TLRs介导的信号通路在马链球菌感染RAW264.7细胞中的作用

Effect of TLRs Mediated Signal Pathway on Streptococcus equi Infection in RAW264.7 Cells

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

[1]	邵永斌, 郑一闯, 张竞怡, 魏琳颖, 田勇, 孙国结, 刘雨然, 罗燕, 谷新利. 益母草碱对LPS诱导小鼠子宫内膜炎的抗炎作用研究[J]. 中国畜牧兽医, 2023, 50(5): 2149-2155.
[2]	付京城, 何俊辉, 仝江, 吴魏, 郭爽, 杨彦宾, 韩莹倩, 王月影, 李和平. HO-1在巨噬细胞中的抗炎和抗氧化作用[J]. 中国畜牧兽医, 2023, 50(2): 807-816.
[3]	谢晶晶, 任梦婷, 范佳慧, 焦亚茹, 韩宗玺, 马得莹. 传染性支气管炎病毒对鸡免疫反应及重组禽β-防御素11和12抗病毒作用的研究[J]. 中国畜牧兽医, 2023, 50(1): 246-259.
[4]	周雅, 张禾垟, 刘琳玲, 李浩东, 郑军军, 王桂武. 梅花鹿成纤维细胞因子受体2基因多态性及其与茸重性状的关联分析[J]. 中国畜牧兽医, 2022, 49(8): 3006-3014.
[5]	陈世钰, 崔帅, 蒋亚君, 鑫婷, 王洋, 郝玉欣, 黄建新, 郭晓宇, 朱鸿飞, 吴佳俊, 贾红. 非洲猪瘟病毒感染后猪外周血淋巴细胞Toll样受体信号通路的lncRNA-mRNA分析[J]. 中国畜牧兽医, 2022, 49(7): 2677-2687.
[6]	刘倩, 郭志廷, 张康, 王学智, 张景艳, 李建喜. 黄芪多糖对巨噬细胞天然免疫调节的研究进展[J]. 中国畜牧兽医, 2022, 49(5): 1746-1756.
[7]	刘雨然, 杨慧莹, 朱妍, 王蒙蒙, 孙国结, 张梦圆, 王燕, 高嘉豪, 连科迅, 谷新利. “乳炎宁”水提物的抗炎作用研究[J]. 中国畜牧兽医, 2022, 49(5): 1879-1887.
[8]	陈毓, 陈晓兰, 杨海峰, 罗军, 张婷, 左海萍, 郑义. 五味止痢合剂对大肠杆菌致小鼠腹泻的治疗作用[J]. 中国畜牧兽医, 2022, 49(5): 2001-2009.
[9]	邱树磊, 陈晓兰, 杨海峰, 陈玉库, 胡元亮, 高永旭, 武彩红. 硒化大蒜多糖对小鼠腹腔巨噬细胞功能的影响[J]. 中国畜牧兽医, 2022, 49(3): 1144-1152.
[10]	买占海, 卢亚宾, 李建龙, 赵璐, 孙丹岚, 佟盼盼, 况玲. 金银花、连翘及其药对对马链球菌马亚种耐药基因和毒力基因的影响[J]. 中国畜牧兽医, 2022, 49(2): 709-717.
[11]	银慧慧, 童艳梅, 曾雪颜, 卢光强, 赵武, 姜源明, 刘伟. 桃金娘果多糖对免疫抑制小鼠免疫功能的影响[J]. 中国畜牧兽医, 2022, 49(2): 731-737.
[12]	朱怡萱, 刘诗芃, 张伟, 李金荣, 彭菲, 林娜, 王语嫣, 刘丹丹. 抗马链球菌马亚种组方加味优选及其体内抑菌作用研究[J]. 中国畜牧兽医, 2022, 49(10): 4085-4096.
[13]	常华, 段纲, 阮谦, 罗倩敏, 余琬玲, 刘清琦, 黄翠琴, 项勋. 红嘴鸥TLR7基因克隆与生物信息学分析[J]. 中国畜牧兽医, 2022, 49(1): 43-52.
[14]	严萍, 林绍青, 李明慧, 孙雪宁, 李健, 黄一帆, 吴异健. 猴头菇多糖对MDRV感染RAW264.7细胞TLR3/TRIF诱导表达及病毒复制的影响[J]. 中国畜牧兽医, 2021, 48(9): 3415-3422.
[15]	张静, 钱英红, 张凯, 吴金迪, 刘博, 毛伟, 曹金山. 金黄色葡菌球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响[J]. 中国畜牧兽医, 2021, 48(8): 3079-3086.