House Dust Mite (HDM)-induced asthma mice were administered recombinant Fgf10 intranasally during activation. Flow cytometry and ELISA were performed to determine type of inflammatory cells and type 2 cytokines levels in bronchoalveolar lavage fluid (BALF). Hematoxylin and eosin (H&E) and periodic acid - Schiff (PAS) staining of lung sections were conducted to evaluate histopathological assessment. Transcriptome profiling was analyzed using RNA-seq, followed by bioinformatics and network analyses to investigate the potential mechanisms of Fgf10 in asthma. RT-qPCR was also used to search for and validate differentially expressed genes in human Peripheral Blood Mononuclear Cells (PBMCs).
Exogenous administration of Fgf10 alleviated HDM-induced inflammation and mucus secretion in lung tissues of mice. Fgf10 also significantly inhibited the accumulation of eosinophils and type 2 cytokines (IL-4, IL-5, and IL-13) in BALF. The PI3K/AKT/NF-κB pathway may mediate the suppressive impact of Fgf10 on the asthma inflammation. Through RNA-seq analysis, the intersection of 71 differentially expressed genes (DEGs) was found between HDM challenge and Fgf10 treatment. GO and KEGG enrichment analyses indicated a strong correlation between the DEGs and different immune response. Immune infiltration analysis predicted the differential infiltration of five types of immune cells, such as NK cells, dendritic cells, monocytes and M1 macrophages. PPI analysis determined hub genes such as Irf7, Rsad2, Isg15 and Rtp4. Interestingly, above genes were consistently altered in human PBMCs in asthmatic patients.
Asthma airway inflammation could be attenuated by Fgf10 in this study, suggesting that it could be a potential therapeutic target.
方法:在激活期间鼻内给予屋尘螨(HDM)诱导的哮喘小鼠重组Fgf10。采用流式细胞术和ELISA检测支气管肺泡灌洗液(BALF)中炎性细胞类型和2型细胞因子水平。对肺切片进行苏木精和伊红(H&E)和高碘酸-希夫(PAS)染色以评估组织病理学评估。使用RNA-seq分析转录组谱分析,随后通过生物信息学和网络分析来研究Fgf10在哮喘中的潜在机制。RT-qPCR也用于搜索和验证人外周血单核细胞(PBMC)中差异表达的基因。
结果:外源性给予Fgf10减轻了HDM诱导的小鼠肺组织炎症和粘液分泌。Fgf10还显著抑制BALF中嗜酸性粒细胞和2型细胞因子(IL-4、IL-5和IL-13)的积累。PI3K/AKT/NF-κB通路可能介导Fgf10对哮喘炎症的抑制作用。通过RNA-seq分析,在HDM攻击和Fgf10处理之间发现了71个差异表达基因(DEGs)的交叉。GO和KEGG富集分析表明DEG与不同免疫应答之间的强相关性。免疫浸润分析预测了五种免疫细胞的差异浸润,如NK细胞,树突状细胞,单核细胞和M1巨噬细胞。PPI分析确定了hub基因,例如Irf7、Rsad2、Isg15和Rtp4。有趣的是,上述基因在哮喘患者的人PBMC中始终发生改变。
结论:在这项研究中,Fgf10可以减轻哮喘气道炎症,这表明它可能是一个潜在的治疗靶点。