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Zheng Libing,Li Yuan,Wang Jun,Pan Ying,Chen Jia,Zheng Weiqiang,Lin Longshan. 2020. Antibacterial and antiparasitic activities analysis of a hepcidin-like antimicrobial peptide from Larimichthys crocea. Acta Oceanologica Sinica, 39(10):129-139
Antibacterial and antiparasitic activities analysis of a hepcidin-like antimicrobial peptide from Larimichthys crocea
大黄鱼抗菌肽hepcidin-like的抗菌和抗寄生虫活性分析
Received:December 24, 2019  
DOI:10.1007/s13131-020-1580-6
Key words:Larimichthys crocea  Cryptocaryon irritans  hepcidin-like  antibacterial activity  antiparasitic activity
中文关键词:  大黄鱼  刺激隐核虫  hepcidin-like  抗菌活性  抗刺激隐核虫活性
基金项目:
Author NameAffiliationE-mail
Zheng Libing State Key Laboratory of Large Yellow Croaker Breeding, Ningde 352103, China
National and Provincial Joint Laboratory of Exploration and Utilization of Marine Aquatic Genetic Resources, School of Marine Science and Technology, Zhejiang Ocean University, Zhoushan 316022, China 
 
Li Yuan Third Institute of Oceanography, Ministry of Natural Resources, Xiamen 361005, China  
Wang Jun State Key Laboratory of Large Yellow Croaker Breeding, Ningde 352103, China  
Pan Ying State Key Laboratory of Large Yellow Croaker Breeding, Ningde 352103, China  
Chen Jia State Key Laboratory of Large Yellow Croaker Breeding, Ningde 352103, China  
Zheng Weiqiang State Key Laboratory of Large Yellow Croaker Breeding, Ningde 352103, China  
Lin Longshan Third Institute of Oceanography, Ministry of Natural Resources, Xiamen 361005, China linlsh2005@126.com 
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Abstract:
      As an economically important marine fish, the large yellow croaker Larimichthys crocea suffered from marine white spot disease caused by the ectoparasite Cryptocaryon irritans in recent years. This disease not only could result in physiological damage, but also lead to secondary bacterial invasion. Reports indicated some AMPs (antimicrobial peptides) were of antiparasitic activity to C. irritans. Hepcidin-like (Lc-HepL) was one of the significant differential expression genes excavated from the transcriptome following a challenge with C. irritans. In this study, we characterized this AMP’s bioactivity based on the levels of mRNA and protein. After challenged by C. irritans, qRT-PCR showed Lc-HepL was significantly upregulated in six tissues, including gill, muscle, liver, head kidney and spleen during theront infection, trophont falling off, and secondary bacterial invasion stages, which implicated a role Lc-HepL played in the immune defense against C. irritans and secondary bacterial infection. Recombinant Lc-HepL (rLc-HepL) was induced and purified successfully. rLc-HepL exhibited antibacterial activity to certain bacteria in a dose- and time-dependent manners. Anti-C. irritans activity was explored for the first time and found it could cause the theronts membrane rupture and contents leakage. These results provided the first evidence that Lc-HepL had strong antiparasitic activity against marine fish ectoparasites C. irritans theronts. Together, data indicated that Lc-HepL might be an important component in the innate immune system against C. irritans and has the potential to be employed in future drug development.
中文摘要:
      近年来,我国重要经济鱼类大黄鱼(Larimichthys crocea)遭受了严重的海水白点病的感染,造成了巨大的经济损失。海水白点病是由体外寄生虫刺激隐核虫(Cryptocaryon irritans)感染引起,不仅能引起被感染鱼类生理的破坏,还能够引发二次细菌感染。有报道指出一些抗菌肽(antimicrobial peptides)具有抗刺激隐核虫的活性。大黄鱼hepcidin-like(命名为Lc-HepL)是从刺激隐核虫感染大黄鱼后的比较转录组中挖掘到的一个差异表达基因。本研究中,在基因和蛋白水平上分析了该抗菌肽的生物活性。刺激隐核虫感染后,qRT-PCR检测到Lc-HepL在鳃、肌肉、肝脏、头肾、肠道和脾脏6种组织中显著上调,并且上调的时间点出现在幼虫感染阶段、滋养体脱落阶段和二次细菌感染阶段,结果显示Lc-HepL在抗刺激隐核虫和二次细菌感染的免疫反应中发挥重要作用。在体外成功诱导并纯化的重组Lc-HepL(rLc-HepL)对某些病原菌具有剂量和时间依赖性的强抗菌活性。本研究首次探究了rLc-HepL的抗刺激隐核虫活性,显微观察结果显示其能引起幼虫细胞膜破裂、内容物外泄,该结果首次提供了大黄鱼hepcidin-like具有强抗刺激隐核虫活性证据。研究数据表Lc-HepL很可能是大黄鱼抗刺激隐核虫感染的一种重要的先天免疫因子,具有应用到未来的药物中的潜力。
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