Previous studies have found that the expression level of Megalobrama amblycephala intelectin (MaINTL) increased significantly post Aeromonas hydrophila infection, and recombinant MaINTL (rMaINTL) protein could activate macrophages and enhance the phagocytosis and killing activity of macrophages. In order to reveal the immune regulatory mechanisms of MaINTL, primary M. amblycephala macrophages were treated with endotoxin-removed rMaINTL and GST-tag proteins, then total RNA were extracted and used for comparative Digital Gene Expression Profiling (DGE). 1247 differentially expressed genes were identified by comparing rMaINTL and GST-tag treated macrophage groups, including 482 up-regulated unigenes and 765 down-regulated unigenes. In addition, eleven randomly selected differentially expressed genes were verified by qRT-PCR, and most of them shared the similar expression patterns as that of DGE results. GO enrichment revealed that the differentially expressed genes were mainly concentrated in the membrane part and cytoskeleton of cellular component, the binding and signal transducer activity of molecular function, the cellular process, regulation of biological process, signaling and localization of biological process, most of which might related with the phagocytosis and killing activity of macrophages. KEGG analysis revealed the activation and involvement of differentially expressed genes in immune related pathways, such as Tumor necrosis factor (TNF) signaling pathway, Interleukin 17 (IL-17) signaling pathway, Toll-like receptor signaling pathway, and NOD like receptor signaling pathway, etc. In these pathways, TNF-ɑ, Activator protein-1 (AP-1), Myeloid differentiation primary response protein MyD88 (MyD88), NF-kappa-B inhibitor alpha (ikBɑ) and other key signaling factors were significantly up-regulated. These results will be helpful to clarify the immune regulatory mechanisms of fish intelectin on macrophages, thus providing a theoretical basis for the prevention and control of fish bacterial diseases.

以往的研究发现，嗜水气单胞菌感染后Megalobrama amblycephala intelectin（MaINTL）的表达水平显着增加，重组MaINTL（rMaINTL）蛋白可以激活巨噬细胞并增强巨噬细胞的吞噬和杀伤活性。为了揭示 MaINTL 的免疫调节机制，初级M. amblcephala巨噬细胞用去除内毒素的 rMaINTL 和 GST 标签蛋白处理，然后提取总 RNA 并用于比较数字基因表达分析 (DGE)。通过比较 rMaINTL 和 GST-tag 处理的巨噬细胞组，鉴定了 1247 个差异表达的基因，包括 482 个上调的 unigenes 和 765 个下调的 unigenes。此外，随机选择的 11 个差异表达基因通过 qRT-PCR 验证，其中大部分与 DGE 结果具有相似的表达模式。GO富集揭示差异表达基因主要集中在细胞成分的膜部分和细胞骨架、分子功能的结合和信号转导活性、细胞过程、生物过程的调控、生物过程的信号传导和定位、其中大部分可能与巨噬细胞的吞噬和杀伤活性有关。KEGG分析揭示了差异表达基因在免疫相关通路中的激活和参与，如肿瘤坏死因子（TNF）信号通路、白细胞介素17（IL-17）信号通路、Toll样受体信号通路和NOD样受体信号通路等。在这些通路中，TNF-ɑ、Activator protein-1（AP-1）、髓系分化初级反应蛋白MyD88（MyD88）、NF-kappa-B抑制剂α（ikBɑ）等关键信号因子显着升高。受监管。这些结果将有助于阐明鱼类智力素对巨噬细胞的免疫调节机制，从而为鱼类细菌性疾病的防治提供理论依据。