Laboratory of Biochemistry and Molecular Biology, School of Marine Sciences, Ningbo University, Ningbo 315211, China.
Laboratory of Biochemistry and Molecular Biology, School of Marine Sciences, Ningbo University, Ningbo 315211, China; Key Laboratory of Applied Marine Biotechnology of Ministry of Education, Ningbo University, Ningbo 315211, China.
Fish Shellfish Immunol. 2018 Nov;82:319-329. doi: 10.1016/j.fsi.2018.08.041. Epub 2018 Aug 18.
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. Upon V. anguillarum infection, 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.
脂联素 (ADP) 作为先天免疫系统的调节剂，在哺乳动物的炎症和代谢紊乱的进展中发挥作用。然而，ADP 在鱼类中的作用还知之甚少。在这里，我们从牙鲆克隆了 ADP 同源物 (PaADP) 的 cDNA 序列。多重序列比对显示 PaADP 具有 ADP 的典型特征。系统进化树分析表明 PaADP 与虹鳟鱼的亲缘关系最为密切。在健康的牙鲆中，PaADP 的转录本在大多数检测到的组织和细胞中均有表达，在脂肪组织中表达水平最高。在鳗弧菌感染后，除脂肪组织外，PaADP 的 mRNA 表达在组织和细胞中均显著上调。随后，在细菌中进行了全长成熟 PaADP（fPaADP）和球状结构域片段（gPaADP）的原核表达和纯化，并产生了抗 PaADP 抗体。Western blot 分析表明，在牙鲆血清中存在包括 fPaADP 和 gPaADP 在内的三种片段。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杀菌作用，对牙鲆 MO/MФ 具有抗炎作用。相比之下，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杀菌作用，对牙鲆 MO/MФ 具有促炎作用。这些结果表明，fPaADP 和 gPaADP 在调节牙鲆 MO/MФ 功能方面发挥着相反的作用。
