State Key Laboratory of Biocontrol, Guangdong Provincial Key Laboratory for Aquatic Economic Animals and Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, PR China.
State Key Laboratory of Biocontrol, Guangdong Provincial Key Laboratory for Aquatic Economic Animals and Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, PR China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266235, PR China; College of Ocean, Hainan University, Haikou, 570228, PR China.
Fish Shellfish Immunol. 2022 Jan;120:102-110. doi: 10.1016/j.fsi.2021.10.024. Epub 2021 Nov 1.
Vibrio parahaemolyticus is an important marine pathogen that cause inflammation even death in teleost. It has brought huge economic losses to aquaculture and serious threats to the sustainable development of marine fisheries. Here, we isolated the DNA, RNA, and total flagellin from V. parahaemolyticus, and obtained the primary spleen and head kidney cells (including leukocytes) from Tetraodon nigroviridis. V. parahaemolyticus DNA, RNA, and total flagellin were used to treat the T. nigroviridis primary cells described above. The results show that the nitric oxide (NO) production and respiratory burst response were significantly induced after stimulation with V. parahaemolyticus total flagellin in T. nigroviridis head kidney and spleen cells. And total flagellin could promote the gene expression and protein production of IL-1β in T. nigroviridis leukocytes. T. nigroviridis TLR5M (TnTLR5M) and TLR5S (TnTLR5S) ORF sequences were obtained as the main recognition receptor for flagellin. Real-time fluorescent quantitative PCR (qRT-PCR) was performed to detect the expression of pattern recognition receptor TnTLR5M and TnTLR5S, the important signal molecule of inflammatory system TnMyD88 and TnTRAF6, and inflammatory cytokines TnIL-1β and TnIFN-γ2. The results show that there were a significant upregulation after challenge with V. parahaemolyticus total flagellin. We further demonstrated that the total flagellin of V. parahaemolyticus could activate the luciferase activity of the NF-κB reporter gene mediated by TnTLR5M. Overall, our results suggest that V. parahaemolyticus total flagellin activated the NO production, respiratory burst response, and inflammatory cytokines expressions, such as TnIL-1β and TnIFN-γ2, through the TnTLR5M-NF-κB signaling pathway in T. nigroviridis.
副溶血弧菌是一种重要的海洋病原体，可引起鱼类的炎症甚至死亡。它给水产养殖业带来了巨大的经济损失，对海洋渔业的可持续发展构成了严重威胁。在这里，我们从副溶血弧菌中分离出 DNA、RNA 和总鞭毛，从叉尾斗鱼中获得了初级脾脏和头部肾脏细胞（包括白细胞）。用副溶血弧菌 DNA、RNA 和总鞭毛处理上述叉尾斗鱼原代细胞。结果表明，副溶血弧菌总鞭毛刺激后，叉尾斗鱼头部肾脏和脾脏细胞的一氧化氮（NO）产生和呼吸爆发反应明显增强。总鞭毛可促进白细胞中 IL-1β的基因表达和蛋白产生。获得了叉尾斗鱼 TLR5M（TnTLR5M）和 TLR5S（TnTLR5S）ORF 序列作为鞭毛的主要识别受体。采用实时荧光定量 PCR（qRT-PCR）检测模式识别受体 TnTLR5M 和 TnTLR5S、炎症系统 TnMyD88 和 TnTRAF6 的重要信号分子以及炎症细胞因子 TnIL-1β和 TnIFN-γ2的表达。结果表明，副溶血弧菌总鞭毛刺激后表达明显上调。我们进一步证明，副溶血弧菌总鞭毛可激活 TnTLR5M 介导的 NF-κB 报告基因的荧光素酶活性。总的来说，我们的结果表明，副溶血弧菌总鞭毛通过 TnTLR5M-NF-κB 信号通路激活了 NO 产生、呼吸爆发反应和炎症细胞因子 TnIL-1β 和 TnIFN-γ2的表达。
