黄条鰤白介素-1β基因克隆及其免疫应答分析  

Two Putative Interleukin-1 beta Molecules Involved in the Immune Response of Seriola aureovittata

作　　者：王琳 王滨[1,2] 徐永江 关长涛[1,2] WANG Lin;WANG Bin;XU Yongjiang;GUAN Changtao(Key Laboratory of Sustainable Development of Marine Fisheries,Ministry of Agriculture and Rural Affairs,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China;Laboratory for Marine Fisheries Science and Food Production Processes,Laoshan Laboratory,Qingdao 266237,China)

机构地区：[1]农业农村部海洋渔业可持续发展重点实验室(中国水产科学研究院黄海水产研究所),山东青岛266071 [2]崂山实验室海洋渔业科学与食物产出过程功能研究室,山东青岛266237

出　　处：《渔业科学进展》2023年第4期64-73,共10页Progress in Fishery Sciences

基　　金：国家重点研发计划项目(2022YFD2401102);青岛海洋科学与技术试点国家实验室“十四五”重大项目(2022QNLM300001-1);中国水产科学研究院基本科研业务费(2020TD51;2020TD47);农业农村部财政专项–海洋渔业生物资源收集与保藏项目;财政部和农业农村部:国家现代农业产业技术体系(CARS-47)共同资助。

摘　　要：白介素-1β是一种典型的促炎细胞因子,参与调控免疫细胞增殖、分化和凋亡等过程。本研究从黄条鰤(Seriola aureovittata)中鉴定到2个白介素-1β分子(分别命名为SaIL-1β1和SaIL-1β2)。SaIL-1β1全长c DNA序列为1292 bp,开放阅读框长度为828 bp,编码275个氨基酸;SaIL-1β2 cDNA序列为1337 bp,开放阅读框长度为960 bp,编码319个氨基酸。SaIL-1β1和SaIL-1β2编码的蛋白均含有IL-1保守的结构域和12个β折叠,具有结构上的保守性。组织表达分布显示,SaIL-1β1在头肾中表达量最高,脾脏和肝脏次之;而SaIL-1β2在鳃中表达量最高,头肾和脾脏次之。脂多糖(LPS)刺激后,SaIL-1β1和SaIL-1β2在头肾和脾脏中的表达量均显著增加。在头肾中,LPS刺激后6 h,SaIL-1β1急剧上升至对照组的10.03倍(P<0.05),随后逐渐回落,在12、24、48、72 h分别为对照组的7.15、4.09、2.71、3.03倍(P<0.05);在刺激后6 h,SaIL-1β2表达量急剧上升至对照组的11.49倍(P<0.05),最后逐渐回落,48h恢复至正常水平,72h下降至对照组的0.29倍(P<0.05)。脾脏中,LPS刺激后6h,SaIL-1β1表达量急剧上升至对照组的6.59倍(P<0.05),随后逐渐回落;SaIL-1β2转录水平表达模式与SaIL-1β2相似。综上,本研究在黄条鰤中鉴定了2种白介素-1β分子,并探讨了其在免疫应答中的表达规律,为研究白介素-1β分子在黄条鰤抗菌免疫中的作用提供了基础。Interleukin-1 beta(IL-1β)is the quintessential pro-inflammatory cytokine,playing important roles in immune cell proliferation,differentiation,and apoptosis.The IL-1β genes have been characterized for many fish species.Unlike mammalian genes,several species of fish possess two IL-1β genes,which may be a consequence of genome duplication in particular fish lineages.Yellowtail kingfish(Seriola aureovittata)is a pelagic marine finfish species,which is an emerging candidate for the aquaculture industry.Therefore,details encompassing the role of IL-1βin the immune response aids a development strategy for economic and efficient aquaculture.In the present study,two novel il-1βmolecules were identified from S.aureovittata(designated as SaIL-1β1 and SaIL-1β2).The full-length cDNA of SaIL-1β1 was 1292 bp with a 828 bp open reading frame,encoding a polypeptide of 275 amino acids,while the full-length cDNA of SaIL-1β2 was 1337 bp with a 960 bp open reading frame,encoding a polypeptide of 319 amino acids.Both SaIL-1βmolecules contain an IL1 domain,12β-sheets,and a C-terminal conserved region,which are IL-1 family signature characters.A phylogenetic analysis revealed the fish IL-1βs clustered together.SaIL-1β1 and IL-1βin Seriola dumerili initially clustered together.However,SaIL-1β2 initially clustered with IL-1β in Trachinotus ovatu.Real-time PCR showed the transcripts of SaIL-1β1 and SaIL-1β2 were present in all the tested tissues,including the head kidney,spleen,liver,gill,heart,stomach,pituitary gland,muscle,and brain.Among them,the SaIL-1β1 transcripts were predominantly in the head kidney,spleen,and liver.The expression of SaIL-1β2 mRNA was predominantly in the gill,head kidney,and spleen.The high expression of SaIL-1β1 and SaIL-1β2 mRNA in the immune related organs implies a potential role in immune regulation.LPS is a pro-inflammatory endotoxin used as a standard immune activating agent.After LPS stimulation,the two SaIL-1βs transcripts were vigorously altered in the head kidney and spleen.SaIL-1

关 键 词：白介素-1Β 免疫应答 细胞因子 黄条鰤 

分 类 号：Q71[生物学—分子生物学]