Abstract：Insulin-like growth factor-1 (IGF-1) is an important endocrine hormone which regulates cell growth and differentiation, and participates in the regulation of immune system.To investigate the mechanisms of IGF-1 in regulating fish immune response to pathogens and explore the regulation mechanism of IGF-1 on fish monocytes/macrophages (MO/MΦ), the present study got Plecoglossus altivelis PaIGF-1 cDNA sequence (GenBank No.MK005188) from transcriptome data.The sequence had 929 bp in length, contained an ORF which encoded a protein of 171 amino acids with the molecular weight of 18.86 kD.The amino acid sequence of PaIGF-1 was relatively conservative, sequence comparison showed that PaIGF-1 shared 77% amino acid sequence identity with Atlantic salmon (Salmo salar).Analysis of qRT-PCR showed that the expression of PaIGF-1 was in a high level in liver, trunk kidney, head kidney and intestine.After infected with Vibrio anguillarum, the expression of PaIGF-1 increased at 8 h in spleen, and increased at 24 h in head kidney, and there was no change in gill and liver.Furthermore, prokaryotic expression system was used to induce the expression and purification of PaIGF-1 recombinant protein (rPaIGF-1), and the mononuclear macrophages were incubated with rPaIGF-1.It was found that rPaIGF-1 increased the phagocytic activity of ayu MO/MΦ and had no effect on bacterial killing.Additionally, the qRT-PCR analysis showed that rPaIGF-1 increased the expression of interleukin-10 (IL-10) and transforming growth factor-β (TGF-β), suppressed the expression of IL-1β and tumor necrosis factor-α (TNF-α) in ayu MO/MФ infected with V.anguillarum.Finally, mononuclear macrophages PaCXCR (C-X-C chemokine receptor) 3.1+ and PaCXCR3.2+ were incubated with rPaIGF-1.It was found that rPaIGF-1 promoted the phosphorylation of PaSTAT3 in PaCXCR3.2+ but had no effect on the phosphorylation levels of PaSTAT1 and PaSTAT3 in PaCXCR3.1+.Inhibition of PaSTAT3 expression by specific siRNA could significantly down-regulate the expression of IL-10 and TGF-β in PaCXCR3.2+ treated with rPaIGF-1 which indicated that rPaIGF-1 might inhibit the inflammatory response of ayu MO/MΦ through activating PaSTAT3.The above research provides a theoretical basis to study PaIGF-1 function and related mechanism in regulating fish immune response to pathogens.
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