Background: NK cells play an important role in immune response, immune surveillance, and metabolism regulation. Therefore, NK cells are involved in the occurrence and development of various diseases, such as infectious diseases, cancer, obesity, and diabetes. IL-25 is a special member of the IL-17 family with anti-inflammatory function. IL-25 can regulate inflammatory response and metabolism via various immune cells; however, the role and regulatory mechanism of IL-25 in NK cells are still unclear. Method: In this study, we investigate the role of IL-25 in NK-cell protein profile via 4D label-free mass spectrum and validate the differential proteins via PRM analysis. In addition, GO analysis, KEGG analysis, and other bioinformatic analysis methods are used to explore the enriched function and signal pathway of differentially expressed proteins. Result and Discussion: The GO and KEGG analyses suggest that IL-25 may affect the processes, such as metabolism, thermogenesis, and oxidative phosphorylation of NK cells. There are 7 downregulated proteins (NCR1, GZMB, PRF1, KLRC1, NDUFA11, LAMTOR5, and IKBIP) and 1 up- -regulated protein (PSMD7) in IL-25-treated NK cells versus the control group for PRM validation. Our results indicate that IL-25 may regulate metabolism and other biological processes via NK cells, which will be beneficial in revealing the role and regulatory mechanisms of IL-25 in NK cells in various diseases. Conclusion: Proteomics combined with bioinformatic analysis will help to mine more information hidden behind mass spectrometry data and lay the foundation for finding clinical biomarkers and mechanisms of diseases.

中文翻译：

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通过 4D 无标记 MS-PRM 进行蛋白质组分析，深入了解 IL-25 在 NK 细胞中的作用和调节机制

背景：NK细胞在免疫反应、免疫监视和代谢调节中发挥着重要作用。因此，NK细胞参与多种疾病的发生和发展，如传染病、癌症、肥胖、糖尿病等。IL-25是IL-17家族的特殊成员，具有抗炎功能。IL-25可以通过多种免疫细胞调节炎症反应和代谢；然而，IL-25在NK细胞中的作用和调控机制仍不清楚。方法：在本研究中，我们通过 4D 无标记质谱研究 IL-25 在 NK 细胞蛋白谱中的作用，并通过 PRM 分析验证差异蛋白。此外，还利用GO分析、KEGG分析等生物信息学分析方法探索差异表达蛋白的富集功能和信号通路。结果与讨论：GO和KEGG分析表明IL-25可能影响NK细胞的代谢、产热和氧化磷酸化等过程。与 PRM 验证的对照组相比，IL-25 处理的 NK 细胞中有 7 个下调的蛋白（NCR1、GZMB、PRF1、KLRC1、NDUFA11、LAMTOR5 和 IKBIP）和 1 个上调的蛋白（PSMD7）。我们的研究结果表明IL-25可能通过NK细胞调节代谢和其他生物过程，这将有利于揭示IL-25在NK细胞中在多种疾病中的作用和调节机制。结论：蛋白质组学结合生物信息分析将有助于挖掘更多隐藏在质谱数据背后的信息，为寻找临床生物标志物和疾病机制奠定基础。