Key Laboratory of Freshwater Aquatic Genetic Resources Ministry of Agriculture, Shanghai Ocean University, Shanghai, China.
Key Laboratory of Freshwater Aquatic Genetic Resources Ministry of Agriculture, Shanghai Ocean University, Shanghai, China; Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai, China; National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China.
Dev Comp Immunol. 2023 Jun;143:104583. doi: 10.1016/j.dci.2022.104583. Epub 2023 Feb 18.
The current study demonstrated that miR-217 modulates inflammation in grass carp (Ctenopharyngodon Idella) infected with Aeromonas hydrophila. Bacterial infection in grass carp causes high levels septicemia, which arises with systemic inflammatory responses. As a result leading to the development of hyperinflammatory state which causes septic shocks and lethality. Based on the current data, TBK1 was confirmed to be the target gene of miR-217 after a successful gene expression profiling or luciferase experiment and miR-217 expression in CIK cells. Furthermore, TargetscanFish6.2 predicted TBK1 as the target gene of miR-217. Quantitative real-time PCR was performed to measure miR-217 expression levels for six immune-related genes and miR-217 regulation in grass carp after A. hydrophila infection in CIK cells. In grass carp CIK cells, the expression of TBK1 mRNA was up-regulated under poly (I: C) stimulation. The transcriptional analysis of the immune-related genes demonstrated that the expression levels of tumor necrosis factor-α (TNF-α), interferon (ifn), interleukin 6 (il-6), interleukin 8 (il-8), and interleukin 12 (il-12) were altered after a successful transfection into the CIK cells, proposing that miRNA regulates immune responses in grass carp. These results provided a theoretical basis and contribute to further studies on the pathogenesis and host defensive system during A. hydrophila infection.
本研究表明，miR-217 可调节受嗜水气单胞菌感染的草鱼的炎症反应。草鱼的细菌感染会导致败血症，败血症会引起全身性炎症反应。因此，导致过度炎症状态，引起败血症休克和死亡率。基于目前的数据，成功进行了基因表达谱或荧光素酶实验以及 CIK 细胞中的 miR-217 表达后，证实 TBK1 是 miR-217 的靶基因。此外，TargetscanFish6.2 预测 TBK1 是 miR-217 的靶基因。在 CIK 细胞中用嗜水气单胞菌感染草鱼后，通过定量实时 PCR 测量了六个免疫相关基因的 miR-217 表达水平和 miR-217 对草鱼的调节作用。在草鱼 CIK 细胞中，TBK1 mRNA 的表达在 poly (I: C) 刺激下上调。免疫相关基因的转录分析表明，肿瘤坏死因子-α (TNF-α)、干扰素 (IFN)、白细胞介素 6 (IL-6)、白细胞介素 8 (IL-8) 和白细胞介素 12 (IL-12) 的表达水平在成功转染到 CIK 细胞后发生改变，这表明 miRNA 调节草鱼的免疫反应。这些结果为嗜水气单胞菌感染期间的发病机制和宿主防御系统的进一步研究提供了理论依据。
