侯庆华,丁旭,卢丹琪

• 1:广东海洋大学海洋与气象学院
• 2:中山大学水生经济动物研究所暨广东省水生经济动物繁殖重点实验室
• 3:西京医院妇产科生殖医学中心

摘要(Abstract)：

构建斜带石斑鱼(Epinephelus coioides)核苷酸结合结构域(Nucleotide-binding domain,NOD)1及2基因的表达载体pEGFP-Nod1和pEGFP-Nod2,与报告基因质粒pNFκB-Luc及内参质粒pRL-TK共转染HEK293T细胞,以脂多糖(LPS)、肽聚糖(PGN)、胞壁酰二肽(MDP)、及二氨基庚二酸(DAP)为细菌类似物代表,以多聚肌苷酸-胞苷酸(Poly I:C)、多聚尿苷酸(PolyU)、多聚脱氧腺苷酸-脱氧胸腺苷酸[Poly(da:dt)]为病毒类似物代表,研究病原类似物刺激后核因子κB(NF-κB)的活化程度。结果表明,斜带石斑鱼Nod2基因可通过NF-κB信号途径识别多种病原成分;Nod1基因可识别DAP,对其他配体呈抑制作用或未通过NF-κB信号通路发挥作用。斜带石斑鱼Nod1及Nod2基因可不同程度地通过NF-κB信号途径识别病原成分。

关键词(KeyWords)： 斜带石斑鱼;NOD1蛋白;NOD2蛋白;病原类似物;核因子κB;双荧光素酶报告系统

Abstract:

Keywords:

基金项目(Foundation): 广东省自然科学基金(2014A030313214);; 现代农业人才支撑计划项目(2016-2020)

作者(Author): 侯庆华,丁旭,卢丹琪

参考文献(References)：

• [1]金伯泉.固有免疫中模式识别受体及其信号转导——当代免疫学中最伟大的发现之一[J].细胞与分子免疫学杂志,2006,22(1):1-3.
• [2]王卫卫,吴谡琦,孙修勤,等.硬骨鱼免疫系统的组成与免疫应答机制研究进展[J].海洋科学进展,2010,28(2):257-265.
• [3]TING J,DUNCAN J A,LEI Y.How the noninflammasome NLRs function in the innate immune system[J].Science,2010,327(5963):286-290.
• [4]CARNEIRO L A,TRAVASSOS L H,PHILPOTT D J.Innate immune recognition of microbes through Nod1 and Nod2:implications for disease[J].Microbes and Infection,2004,6(6):609-616.
• [5]GIRARDIN S E,BONECA I G,CARNEIRO L,et al.Nod1 detects a unique muropeptide from Gram-negative bacterial peptidoglycan[J].Science,2003,300(5625):1584-1587.
• [6]MO J Y,BOYLE J P,HOWARD C B,et al.Pathogen sensing by nucleotide-binding oligomerization domain-containing protein 2(NOD2)is mediated by direct binding to muramyl dipeptide and ATP[J].The Journal of Biological Chemistry,2012,287(27):23057-23067.
• [7]CHAMAILLARD M,HASHIMOTO M,HORIE Y et al.An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid[J].Nature Immunology,2003,4(7):702-707.
• [8]TATTOLI I,TRAVASSOS L H,CARNEIRO L A et al.The Nodosome:Nod1 and Nod2 control bacterial infections and inflammation[J].Seminars in Immunopathology,2007,29(3):289-301.
• [9]VIALA M,SANSONETTI P,PHILPOTT D J.Nods and‘intracellular’innate immunity[J].Comptes Rendus Biologies,2004,327(6):551-555.
• [10]LAROUI H,YAN Y T,NARUI Y,et al.L-Ala-gamma-D-Glu-meso-diaminopimelic Acid(DAP)interacts directly with leucine-rich region domain of nucleotide-binding oligomerization domain 1,increasing phosphorylation activity of receptor-interacting serine/threonine-protein kinase 2 and its interaction with nucleotide-binding oligomerization domain 1[J].The Journal of Biological Chemistry,2011,286:31003-31013.
• [11]CHANG M X,WANG T H,NIE P,et al.Cloning of two rainbow trout nucleotide-binding oligomerization domain containing 2(NOD2)splice variants and functional characterization of the NOD2 effector domains[J].Fish&Shellfish Immunology,2012,30(1):118-127.
• [12]CHEN W Q,XU Q Q,CHANG M X,et al.Molecular characterization and expression analysis of nuclear oligomerization domain proteins NOD1 and NOD2 in grass carp Ctenopharyngodon idella[J].Fish&Shellfish Immunology,2010,28(1):18-29.
• [13]KOBAYASHI K,INOHARA N,HERNANDEZ L D,et al.RICK/Rip2/CARDIAK mediates signalling for receptors of the innate and adaptive immune systems[J].Nature,2002,416(6877):194-199.
• [14]NEMBRINI C,KISIELOW J,SHAMSHIEV A T,et al.The kinase activity of Rip2 determines its stability and consequently Nod1-and Nod2-mediated immune responses[J].The Journal of Biological Chemistry,2009,284(29):19183-19188.
• [15]SWAIN B,BASU M,SAMANTA M.Molecular cloning and characterization of nucleotide binding and oligomerization domain-1(NOD1)receptor in the Indian Major Carp,rohu(Labeo rohita),and analysis of its inductive expression and down-stream signalling molecules following ligands exposure and Gram-negative bacterial infections[J].Fish&Shellfish Immunology,2012,32(5):899-908.
• [16]HOU Q H,YI S B,DING X,et al.Differential expression analysis of nuclear oligomerization domain proteins NOD1 and NOD2 in orange-spotted grouper(Epinephelus coioides)[J].Fish&Shellfish Immunology,2012,33:1102-1111.
• [17]MONIE T P.NLR activation takes a direct route[J].Trends in Biochemical Sciences,2013,38(3):131-139.
• [18]BARNICH N,AGUIRRE J E,REINECKER H C,et al.Membrane recruitment of NOD2 in intestinal epithelial cells is essential for nuclear factor-{kappa}B activation in muramyl dipeptide recognition[J].The Journal of Biological Chemistry,2005,170(1):21-26.
• [19]BOYLE J P,PARKHOUSE R,MONIE T P.Insights into the molecular basis of the NOD2 signalling pathway[J].Open Biology,2014,4(12):1-12.
• [20]MONIE T P.NLR activation takes a direct route[J].Trends in Biochemical Sciences,2013,38(3):131-139.
• [21]CORREA R G,MILUTINOVIC S,REED J C.Roles of NOD1(NLRC1)and NOD2(NLRC2)in innate immunity and inflammatory diseases[J].Bioscience Reports,2012,32(6):597-608.
• [22]FRANCHI L,PARK J H,SHAW M H,et al.Intracellular NOD-like receptors in innate immunity,infection and disease[J].Cellular Microbiology,2008,10(1):1-8.
• [23]GIRARDIN S E,TOURNEBIZE R,MAVRIS M,et al.CARD4/Nod1 mediates NF-kappa B and JNK activation by invasive Shigella flexneri[J].EMBO reports,2001,2(8):736-742.
• [24]KAPARAKIS-LIASKOS M.The intracellular location,mechanisms and outcomes of NOD1 signaling[J].Cytokine,2015,74(2):207-212.
• [25]INOHARA N,OGURA Y,CHEN F F,et al.Human Nod1 confers responsiveness to bacterial lipopolysaccharides[J].The Journal of Biological Chemistry,2001,276(4):2551-2554.
• [26]Allison C C,Kufer T A,Kremmer E,et al.Helicobacter pylori induces MAPK phosphorylation and AP-1activation via a NOD1-dependent mechanism[J].The Journal of Immunology,2009,183(12):8099–8109.
• [27]OPITZ B,PüSCHEL A,BEERMANN W,et al.Listeria monocytogenes activated p38 MAPK and induced IL-8secretion in a nucleotide-binding oligomerization domain 1-dependent manner in endothelial cells[J].The Journal of Immunology,2006,176(1):484-490.
• [28]KANNEGANTI T D,LAMKANFI M,Nú?EZ G.Intracellular NOD-like receptors in host defense and disease[J].Immunity,2007,27(4):549-559.
• [29]SHAW M H,REIMER T,KIM Y G,et al.NOD-like receptors(NLRs):bona fide intracellular microbial sensors[J].Current Opinion in Immunology,2008,20(4):377-382.