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沈海钰, 周燕, 周前进, 李明云, 陈炯. 2020: 大弹涂鱼白细胞介素-34(IL-34)通过集落刺激因子-1受体1 (CSF-1R1) 调节单核/巨噬细胞功能的研究. 动物学研究, 41(2): 123-137. DOI: 10.24272/j.issn.2095-8137.2020.026
引用本文: 沈海钰, 周燕, 周前进, 李明云, 陈炯. 2020: 大弹涂鱼白细胞介素-34(IL-34)通过集落刺激因子-1受体1 (CSF-1R1) 调节单核/巨噬细胞功能的研究. 动物学研究, 41(2): 123-137. DOI: 10.24272/j.issn.2095-8137.2020.026
Hai-Yu Shen, Yan Zhou, Qian-Jin Zhou, Ming-Yun Li, Jiong Chen. 2020: Mudskipper interleukin-34 modulates the functions of monocytes/macrophages via the colony-stimulating factor-1 receptor 1. Zoological Research, 41(2): 123-137. DOI: 10.24272/j.issn.2095-8137.2020.026
Citation: Hai-Yu Shen, Yan Zhou, Qian-Jin Zhou, Ming-Yun Li, Jiong Chen. 2020: Mudskipper interleukin-34 modulates the functions of monocytes/macrophages via the colony-stimulating factor-1 receptor 1. Zoological Research, 41(2): 123-137. DOI: 10.24272/j.issn.2095-8137.2020.026

大弹涂鱼白细胞介素-34(IL-34)通过集落刺激因子-1受体1 (CSF-1R1) 调节单核/巨噬细胞功能的研究

Mudskipper interleukin-34 modulates the functions of monocytes/macrophages via the colony-stimulating factor-1 receptor 1

  • 摘要: 新型细胞因子白细胞介素-34(interleukin-34,IL-34)通过与集落刺激因子-1受体(the colony-stimulating factor-1 receptor,CSF-1R)结合,在先天免疫和炎症中发挥重要作用。目前,关于鱼类IL-34功能的研究还非常有限。本研究鉴定了大弹涂鱼(Boleophthalmus pectinirostris)的IL-34(BpIL-34),在序列和结构与其它已知的IL-34相似,与石斑鱼(Epinephelus coioides)的同源关系最近。mRNA水平的研究发现,BpIL-34能组成型表达于大弹涂鱼的各个组织,以大脑表达水平最高;迟缓爱德华氏菌(Edwardsiella tarda)感染能上调各组织的表达。本研究表达并纯化了BpIL-34的成熟肽(rBpIL-34),制备了抗体。肽N-糖苷酶F酶解发现单核/巨噬细胞的BpIL-34被N-糖基化。体外实验发现rBpIL-34能够增强单核/巨噬细胞的吞噬和杀菌活性,促进肿瘤坏死因子α(BpTNF-α)和BpIL-1β等促炎细胞因子的表达。RNA干扰实验结果表明,敲低大弹涂鱼CSF-1R1(BpCSF-1R1)后,rBpIL-34介导的单核/巨噬细胞功能的强化表现被显著抑制;BpCSF-1R2不具备此功能。综上,BpIL-34能够通过BpCSF-1R1调节大弹涂鱼单核/巨噬细胞的功能。

     

    Abstract: Interleukin-34 (IL-34) is a novel cytokine that plays an important role in innate immunity and inflammatory processes by binding to the colony-stimulating factor-1 receptor (CSF-1R). However, information on the function of IL-34 in fish remains limited. In the present study, we identified an IL-34 homolog from mudskippers (Boleophthalmus pectinirostris). In silico analysis showed that the mudskipper IL-34 (BpIL-34) was similar to other known IL-34 variants in sequence and structure and was most closely related to an orange-spotted grouper (Epinephelus coioides) homolog. BpIL-34 transcripts were constitutively expressed in various tissues, with the highest level of expression found in the brain. Edwardsiella tarda infection significantly up-regulated the mRNA expression of BpIL-34 in the mudskipper tissues. The recombinant mature BpIL-34 peptide (rBpIL-34) was purified and used to produce anti-rBpIL-34 IgG. Western blot analysis combined with PNGase F digestion revealed that native BpIL-34 in monocytes/macrophages (MOs/MФs) was N-glycosylated. In vitro, rBpIL-34 treatment enhanced the phagocytotic and bactericidal activity of mudskipper MOs/MФs, as well as the mRNA expression of pro-inflammatory cytokines like tumor necrosis factor α (BpTNF-α) and BpIL-1β in these cells. Furthermore, the knockdown of mudskipper CSF-1R1 (BpCSF-1R1), but not mudskipper BpCSF-1R2, significantly inhibited the rBpIL-34-mediated enhanced effect on MO/MФ function. In conclusion, our results indicate that mudskipper BpIL-34 modulates the functions of MOs/MФs via BpCSF-1R1.

     

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