Full‐length and a smaller globular fragment of adiponectin have opposite roles in regulating monocyte/macrophage functions in ayu, Plecoglossus altivelis
Full‐length and a smaller globular fragment of adiponectin have opposite roles in regulating monocyte/macrophage functions in ayu, Plecoglossus altivelis
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DOI：
10.1016/j.fsi.2018.08.041
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发表时间：
2018-11
影响因子：
4.7
通讯作者：
He Liu;Xin-Jiang Lu;Jiong Chen-
中科院分区：
文献类型：
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作者：
He Liu;Xin-Jiang Lu;Jiong Chen-
脂联素（ADP）是先天免疫系统的调节剂，在哺乳动物炎症和代谢紊乱的进展中起作用。然而，ADP在鱼类中的作用知之甚少。本研究克隆了香鱼ADP同源基因（PaADP）的cDNA序列。多序列比对显示PaADP具有ADP的典型特征。系统进化树分析表明，PaADP与虹鳟的亲缘关系最近。在健康香鱼中，PaADP在大部分组织和细胞中均有表达，在脂肪组织中表达量最高。五.鳗鲡感染后，除脂肪组织外，其它组织和细胞中PaADP的mRNA表达均显著上调。随后，在细菌中原核表达全长成熟PaADP（fPaADP）和球状结构域片段（gPaADP）并纯化，并产生抗PaADP抗体。Western blot分析表明，香鱼血清中存在fPaADP和gPaADP三个片段。重组fPaADP（rfPaADP）通过上调抗炎细胞因子的表达、下调促炎细胞因子的表达、抑制吞噬作用和随后的细菌杀伤作用，对香鱼MO/M具有抗炎作用。与此相反，重组gPaADP（rgPaADP）通过上调促炎细胞因子的表达，下调抗炎细胞因子的表达，增强吞噬作用和随后的细菌杀伤作用，对香鱼MO/M.这些结果提示fPaADP和gPaADP在香鱼MO/M β功能的调节中具有相反的作用。
Adiponectin (ADP), a regulator of the innate immune system, plays a role in the progression of inflammation and metabolic disorders in mammals. However, the role of ADP in fish is poorly understood. Here, we cloned the cDNA sequence of a ADP homolog (PaADP) gene from ayu. Multiple sequence alignment revealed that PaADP presented typical characteristics of ADPs. Phylogenetic tree analysis showed that PaADP was most closely related to that of rainbow trout. In healthy ayu, the transcripts of PaADP were detected in most of the tested tissues and cells, with the highest level in the adipose tissue. UponV. anguillaruminfection, the mRNA expression of PaADP was significantly up-regulated in the tissues and cells except adipose tissue. Subsequently, the full-length mature PaADP (fPaADP) and the globular domain fragment (gPaADP) were prokaryotically expressed in bacteria and purified, and anti-PaADP antibodies were produced. Western blot analysis revealed that three fragments including fPaADP and gPaADP were existed in ayu serum. The recombinant fPaADP (rfPaADP) had an anti-inflammatory effect on ayu MO/MФ by upregulating anti-inflammatory cytokine expressions, downregulating pro-inflammatory cytokine expressions, inhibiting the phagocytosis and subsequent bacterial killing. In contrast, the recombinant gPaADP (rgPaADP) presented a pro-inflammatory effect on ayu MO/MФ by upregulating pro-inflammatory cytokine expression, downregulating anti-inflammatory cytokine expressions, enhancing the phagocytosis and subsequent bacterial killing. These results suggested that fPaADP and gPaADP have opposite roles in the regulation of MO/MФ functions in ayu.