Sun Yulong, Kuang Yuanyuan, Zuo Zhuo, Zhang Jin, Ma Xiaolong, Xing Xiaoyu, Liu Lingyi, Miao Yuchen, Ren Tao, Li Hui, Mei Qibing
Key Laboratory for Space Biosciences & Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi Province, 710072, China.
Key Laboratory for Space Biosciences & Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi Province, 710072, China.
Peptides. 2021 Feb;136:170469. doi: 10.1016/j.peptides.2020.170469. Epub 2020 Dec 9.
Neuropeptide FF (NPFF) is a neuropeptide that modulates various physiological processes. The regulatory role of NPFF in the immune and inflammatory response is currently being revealed. However, the effect of NPFF at the transcriptome level in macrophages has not been fully elucidated. Here, the impact of NPFF on gene expression at the transcriptome level of RAW 264.7 cells was investigated by RNA-seq. RAW 264.7 macrophages were treated with NPFF (1 nM) for 18 h, followed by RNA-seq examination. Differentially expressed genes (DEGs) were acquired, followed by GO, KEGG, and PPI analysis. A total of eight qPCR-verified DEGs were obtained. Next, three-dimensional models of the eight hub proteins were constructed by using homology modeling with Modeller (9v23). Finally, molecular dynamics simulation (300 ns) was performed with GROMACS 2018.2 to investigate the structural characteristics of these hub proteins. NPFF had no detectable effect on the morphology of RAW264.7 cells. A total of 211 DEGs were acquired, and an enrichment study demonstrated that the immune response-related pathway was significantly inhibited by NPFF. Moreover, the molecular dynamics optimized-protein models of the hub proteins were obtained. Collectively, NPFF inhibited the expression of immune-related genes in RAW 264.7 cells at the transcriptome level, which suggested a negative relationship between NPFF and this set of immune-related genes in RAW 264.7 macrophages. Therefore, our data may provide direct evidence of the role of NPFF in peripheral or central inflammatory diseases.
神经肽FF（NPFF）是一种调节多种生理过程的神经肽。NPFF在免疫和炎症反应中的调节作用目前正在被揭示。然而，NPFF在巨噬细胞转录组水平上的作用尚未完全阐明。在此，通过RNA测序研究了NPFF对RAW 264.7细胞转录组水平基因表达的影响。用NPFF（1 nM）处理RAW 264.7巨噬细胞18小时，随后进行RNA测序检查。获得差异表达基因（DEG），然后进行基因本体论（GO）、京都基因与基因组百科全书（KEGG）和蛋白质-蛋白质相互作用（PPI）分析。共获得8个经qPCR验证的DEG。接下来，使用Modeller（9v23）通过同源建模构建这8个枢纽蛋白的三维模型。最后，使用GROMACS 2018.2进行分子动力学模拟（300纳秒）以研究这些枢纽蛋白的结构特征。NPFF对RAW264.7细胞的形态没有可检测到的影响。共获得211个DEG，富集研究表明免疫反应相关途径受到NPFF的显著抑制。此外，还获得了枢纽蛋白的分子动力学优化蛋白模型。总的来说，NPFF在转录组水平上抑制了RAW 264.7细胞中免疫相关基因的表达，这表明NPFF与RAW 264.7巨噬细胞中的这组免疫相关基因之间存在负相关关系。因此，我们的数据可能为NPFF在周围或中枢炎症性疾病中的作用提供直接证据。
