Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, 266003, Qingdao, Shandong, China.
Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, 266003, Qingdao, Shandong, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, 266237, Qingdao, Shandong, China.
Fish Shellfish Immunol. 2018 Apr;75:327-335. doi: 10.1016/j.fsi.2018.02.021. Epub 2018 Feb 9.
Being a key adaptor protein in NOD1/2 and NF-κB signaling pathways, receptor-interacting serine/threonine kinase 2 (RIP2) plays an important role in innate immune response in vertebrates. In this study, we identified and characterized the Paralichthys olivaceus RIP2 gene (PoRIP2). Phylogenetic, alignment, and genomic analysis of PoRIP2 were conducted to determine its conservation and evolutionary relationship with other RIP2 in vertebrates. qRT-PCR results showed that the expression of PoRIP2 was high in the spleen and head kidney. Meanwhile, embryonic development expression profile revealed that it was high in the early developmental stages and hatching stage. In vivo, we examined the expression pattern in different tissues after being challenged with Edwardsiella tarda. PoRIP2 was up-regulated in tissues at different time points. In vitro, the expression of PoRIP2 was also increased after treatment with Poly I:C, PGN, and E. tarda. Transfection and overexpression experiments indicated that PoRIP2 was located in the cytoplasm of the FG-9307 cell line. The pro-inflammatory cytokines, IL-1β, IL-6, and IL-8, could be activated and up-regulated by PGN stimulation in PoRIP2 overexpressed cells. The inhibitory action was obvious in PoRIP2 overexpressed cells, and the quantity of E. tarda decreased. These findings highlight the important role of PoRIP2 in regulating innate immune in P. olivaceus. Our results indicated that PoRIP2 might be involved in immune response and the activation of the NF-κB signaling pathways. Our study can improve the knowledge on the immune system of fish and provide a theoretical basis for the study of prevention and treatment of fish diseases.
作为 NOD1/2 和 NF-κB 信号通路的关键衔接蛋白，受体相互作用丝氨酸/苏氨酸激酶 2（RIP2）在脊椎动物的固有免疫反应中发挥着重要作用。本研究鉴定并表征了牙鲆 RIP2 基因（PoRIP2）。对 PoRIP2 的系统进化、比对和基因组分析，以确定其与其他脊椎动物 RIP2 的保守性和进化关系。qRT-PCR 结果显示，PoRIP2 在脾脏和头肾中表达量较高。同时，胚胎发育表达谱显示其在早期发育阶段和孵化阶段表达量较高。在体内，我们检测了受到迟缓爱德华氏菌刺激后不同组织中的表达模式。PoRIP2 在不同时间点的组织中均上调。在体外，用 Poly I:C、PGN 和 E. tarda 处理后，PoRIP2 的表达也增加。转染和过表达实验表明，PoRIP2 位于 FG-9307 细胞系的细胞质中。PGN 刺激可激活和上调 PoRIP2 过表达细胞中的促炎细胞因子 IL-1β、IL-6 和 IL-8。PoRIP2 过表达细胞中的抑制作用明显，E. tarda 的数量减少。这些发现强调了 PoRIP2 在调控牙鲆固有免疫中的重要作用。我们的结果表明，PoRIP2 可能参与了免疫反应和 NF-κB 信号通路的激活。我们的研究可以提高对鱼类免疫系统的认识，并为鱼类疾病的防治研究提供理论依据。
Zotero 插件