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香鱼akirin1基因的解析及其对单核巨噬细胞功能的调控

The Characterization of Ayu(Plecoglossus altivelis) akirin1 Gene and Regulation of Function in Monocytes Macrophages
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摘要 Akirin是一个高度保守的核因子,与动物的免疫功能密切相关。本研究克隆获得香鱼(Plecoglossus altivelis)akirin1的c DNA序列(Paakirin1),该基因包含一个549 bp的开放阅读框,编码182个氨基酸(Gen Bank登录号:KP278589)。Paakirin1与虹鳟(Oncorhynchus mykiss)akirin1的氨基酸序列同源性最高,达到86%,而且含有核定位序列。系统进化树分析表明,鱼类akirin1聚为一大簇,Paakirin1和大西洋鲑(Salmo salar)、虹鳟、红点鲑(Salvelinus alpinus)的akirin1聚类在一起。半定量PCR结果显示,Paakirin1主要在头肾、脾、肠和单核巨噬细胞表达。Paakirin1 m RNA在鳗弧菌(Vibrio anguillarum)感染后的香鱼头肾和脾组织中表达量显著上调(P〈0.05)。单核巨噬细胞在多种免疫刺激后Paakirin1 m RNA也上调(P〈0.05)。RNA干扰敲低Paakirin1在单核/巨噬细胞的表达,并不影响单核巨噬细胞在鳗弧菌感染后的白细胞介素-1β(interleukin-1β,IL-1β)和肿瘤坏死因子(tumor necrosis factor,TNF)表达,但是显著下调细胞向重组CXC趋化因子配体8(CXCL8)蛋白的趋化(P〈0.05)。综上,Paakirin1的表达与免疫刺激密切相关,且调控单核巨噬细胞的趋化功能。本研究为进一步了解鱼类的单核巨噬细胞功能和调控提供理论依据和参考。 Akirin,a conserved nuclear factor,is associating with immune functions in animal via regulating cytokine transcription and immune cell chemotaxis.Here,an akirin1 c DNA sequence(Paakirin1) from ayu(Plecoglossus altivelis) was cloned.The Paakirin1 was 1817 bp in length encoded a 182 aa polypeptide with a molecular weight of 20.68 k D(Gen Bank accession No.KP278589).Sequence comparisons for similarity showed that Paakirin1 was highly similar to akirin1 in fish,such as,rainbow trout(Oncorhynchus mykiss)akirin1(86%),followed by Arctic char(Salvelinus alpinus) and Atlantic salmon(Salmo salar) akirin1(85%).Multiple sequence alignment analysis showed that Paakirin1 possessed nuclear localization signal.Phylogenetic analysis showed that fish akirin1 genes grouped tightly into one big cluster.Paakirin1 clustered together with the akirin1 s of salmonidae family.Semi-quantitative PCR data showed that Paakirin1 was mainly expressed in head-kidney,spleen,intestine,and monocytes macrophages.After Vibrio anguillaruminfection,Paakirin1 m RNA was up-regulated in head-kidney and spleen,while down-regulated in intestine.Furthermore,Paakirin1 m RNA was also up-regulated in monocytes macrophages after lipopolysaccharide(LPS) or polyinosinic-polycytidylic acid(Poly I:C) treatment.After Paakirin1 si RNA treatment in monocytes macrophages,the m RNA expression of interleukin-1β(IL-1β) and tumor necrosis factor(TNF) induced by V.anguillarum was unaltered,while its chemotaxis toward to recombinant CXC chemokine ligand 8(CXCL8)protein was down-regulated.In summary,Paakirin1 alters its expression after infection and mediates chemotaxis of ayu monocytes macrophages.The research provides a theoretical basis and reference for the further investigation of function and regulation of monocytes macrophages in fish.
作者 林凡佳 裘天休 陆新江 陈炯 李明云 LIN Fan-Jia,QIU Tian-Xiu, LU Xin-Jiang, CHEN Jiong, LI Ming-Yun( 1 School of Marine Sciences, Ningbo University, Ningbo 315211, China; 2 College of Science & Technology, Ningbo University, Ningbo 315211, China)
出处 《农业生物技术学报》 CAS CSCD 北大核心 2015年第10期1350-1359,共10页 Journal of Agricultural Biotechnology
基金 国家自然科学面上基金资助项目(No.31472300) 高等学校博士学科点专项科研基金(No.20113305120001)
关键词 akirin 香鱼单核巨噬细胞 鳗弧菌 RNA干扰 akirin Ayu Monocytes macrophages Vibrio anguillarum RNA interference
作者简介 通讯作者,lxj711043@163.com
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