Neuropeptide FF-related gene in fish (Larimichthys polyactis): identification, characterization, and potential anti-inflammatory function

文献类型： 外文期刊

第一作者： Xiao, Ran

作者： Xiao, Ran;Guo, Zuting;Zheng, Li-bing;Huang, Wei;Chi, Chang-feng;Lou, Bao

作者机构：

关键词： Larimichthys polyactis; Neuropeptide FF; G-protein coupled receptor; Nitric oxide

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.742； 五年影响因子：2.702 ）

ISSN： 0301-4851

年卷期： 2022 年 49 卷 7 期

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收录情况： SCI

摘要： Background Neuropeptide FF (NPFF), an octapeptide of the RFamide-related peptides (FaRPs), is involved in regulatory function in various biological processes. The regulatory role of NPFF in the immune and inflammatory response was currently being revealed. Methods Neuropeptide FF-related gene (termed LpNPFF) and its two receptors, NPFF receptor 1 (LpNPFFR1) and NPFF receptor 2 (LpNPFFR2) were identified by PCR and Semi-quantitative RT-PCR assay. Effect of LpNPFF on the production of nitric oxide (NO) in macrophage RAW264.7 cell was divided into PBS group, lipopolysaccharide (LPS) group, LPS treated with LpNPFF group, and LPS treated with LpNPFF and receptor antagonist RF9 group. Then specimens were measured by color reaction at 570 nm absorbance value. Results Sequence analysis showed that LpNPFF cDNA consists of 835 nucleotides with a 5 '- untranslated region (UTR) of 150 base pair (bp), an open reading frame (ORF) of 384 bp and a 3 '-UTR of 300 bp (Accession No. MT012894). The ORF encodes 127 amino acid (aa) residues with a hydrophobic signal peptide at N-terminus and two presumptive peptides with -PQRFa structure, LpNPFF (1) and LpNPFF (2). LpNPFFR1 and LpNPFFR2 encode 427 and 444 aa residues respectively, which both have seven hydrophobic TMDs and identified as G protein coupled receptors (GPCRs). Results of tissue distribution showed that LpNPFF and receptors were highly expressed in the brain and gonad. Furtherly, in vitro assay found LpNPFF could inhibit NO production in RAW 264.7 macrophages under inflammatory stress with LPS, while its receptor antagonist RF9 caused the evoke of NO generation. Conclusions These results contribute to the further study of neuropeptide evolution in marine organisms, and also provide a new research idea for exploring the related functions of NPFF gene.

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