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过氧化物酶 4 在克氏原螯虾抗氧化防御和免疫反应中的重要作用。

Essential role of the peroxiredoxin 4 in Procambarus clarkii antioxidant defense and immune responses.

机构信息

School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, PR China.

School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, PR China.

出版信息

Fish Shellfish Immunol. 2018 Apr;75:216-222. doi: 10.1016/j.fsi.2018.01.042. Epub 2018 Feb 1.

Abstract

Peroxiredoxin (Prx) family members play a key role in host defense against oxidative stress, and modulate immune responses following microbial infection. Here, we cloned and characterized Procambarus clarkii Prx4 (Peroxiredoxin 4) cDNA, a regulator of oxidative stress and its expression analysis upon lipopolysaccharide (LPS) and polyriboinosinic polyribocytidylic acid (Poly I:C) infection. The cDNA fragment of PcPrx4 was 744 bp in length, encoding a putative protein of 248 amino acid residues. Real-time quantitative reverse transcription-PCR (qRT-PCR) analysis showed that the PcPrx4 was expressed in all the examined tissues, and it was highest in the hepatopancreas followed by the hemocytes and gill. The challenge with LPS and Poly I:C significantly up-regulated the expression of PcPrx4 in hepatopancreas, hemocytes and gill when compared with the control. Recombinant PcPrx4 protein was used to investigate the antioxidant function in vitro by mixed-function oxidase assay. The results demonstrated a dose-dependent inhibition of DNA damage by rPcPrx4 protein. Altogether, our results imply that PcPrx4 is implicated in defense against microbial pathogens and oxidants in P. clarkii.

摘要

过氧化物酶(Prx)家族成员在宿主抵御氧化应激方面发挥着关键作用,并调节微生物感染后的免疫反应。在这里,我们克隆并鉴定了 Procambarus clarkii Prx4(过氧化物酶 4)cDNA,它是一种氧化应激调节剂,并且在脂多糖(LPS)和聚肌苷酸-聚胞苷酸(Poly I:C)感染后对其表达进行了分析。PcPrx4 的 cDNA 片段长 744bp,编码一个由 248 个氨基酸残基组成的假定蛋白。实时定量反转录 PCR(qRT-PCR)分析表明,PcPrx4 在所有检测的组织中均有表达,在肝胰腺中表达最高,其次是血细胞和鳃。与对照组相比,LPS 和 Poly I:C 的刺激显著上调了肝胰腺、血细胞和鳃中 PcPrx4 的表达。用混合功能氧化酶测定法体外研究重组 PcPrx4 蛋白的抗氧化功能。结果表明 rPcPrx4 蛋白对 DNA 损伤具有剂量依赖性抑制作用。总之,我们的研究结果表明,PcPrx4 参与了克氏原螯虾对微生物病原体和氧化剂的防御。

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