miR-144 affects the immune response and activation of inflammatory responses in Cynoglossus semilaevis by regulating the expression of CsMAPK6

• Published in: Fish & shellfish immunology Vol. 149; p. 109578
• Main Authors: Zheng, Guiliang, Sun, Siqi, Zhang, Guosong, Liang, Xia
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.06.2024
• Subjects: Animals; Chinese tongue sole; Cytokines - genetics; Cytokines - immunology; Cytokines - metabolism; Fish Diseases - immunology; Fish Proteins - genetics; Fish Proteins - immunology; Flatfishes - genetics; Flatfishes - immunology; Gene Expression Regulation - immunology; Immune regulation; Immunity, Innate - genetics; Inflammation - genetics; Inflammation - immunology; Inflammation - veterinary; MAPK6; MicroRNAs - genetics; MicroRNAs - immunology; miR-144; Vibrio - physiology; Vibrio Infections - immunology; Vibrio Infections - veterinary; Immune regulation; Chinese tongue sole; MAPK6; miR-144
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2024.109578

MicroRNAs are increasingly recognized for their pivotal role in the immune system, yet the specific regulatory functions of fish-derived microRNAs remain largely unexplored. In this research, we discovered a novel miRNA, Cse-miR-144, in the Chinese tongue sole (Cynoglossus semilaevis), characterized by a 73-base pair precursor and a 21-nucleotide mature sequence. Our findings revealed that the expression of Cse-miR-144 was notably inhibited by various Vibrio species. Utilizing bioinformatics and dual-luciferase assay techniques, we established that the pro-inflammatory cytokine gene CsMAPK6 is a direct target of Cse-miR-144. Subsequent in vitro and in vivo western blotting analyses confirmed that Cse-miR-144 can effectively reduce the protein levels of CsMAPK6 post-transcriptionally. Moreover, CsMAPK6 is known to be involved in the activation of the Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-kB). Additional investigations using qPCR and ELISA demonstrated that suppression of Cse-miR-144 leads to an upsurge in the liver mRNA levels of various immune genes (including MYD88, TRAF6, NF-κB, TRAF2, TRAF3, and TNF), alongside a marked increase in the production and secretion of pro-inflammatory cytokines (IL-1β, IL-6, and IL-8) in the bloodstream of C. semilaevis. These findings collectively underscore the potential of Cse-miR-144 as a key inhibitor of CsMAPK and its crucial role in modulating the immune and inflammatory responses in teleost fish. Compared to the siRNA, miRNA is a better tool in controlling the expression of target gene with a lower cost. •A novel Cse-miR-144 with potential regulatory impacts on the infection of Vibrio by targeting CsMAPK6 gene was analysis.•Cse-miR-144 has been shown effect on the production of pro-inflammatory cytokines in by downregulating CsMAPK6.•Cse-miR-144 are more efficient than siRNAs for expression control of CsMAPK6 gene and immune response modulation.