【摘要】 目的 运用机器学习方法鉴定过敏体质诊断标记物和关键免疫细胞,探讨过敏康调节过敏体质的作用机制。方法 分析基因表达综合数据库中多种过敏性疾病患者组成的过敏体质人群相较于正常人群的差异表达基因(DEGs),WGCNA算法用于鉴定过敏体质相关基因,并进行蛋白质-蛋白质相互作用、GSEA、GO与KEGG富集分析。结合机器学习算法(LASSO回归和SVM-RFE)鉴定过敏体质诊断标记物,并运用CIBERSORT鉴定计算22种免疫细胞浸润程度,分析诊断标记物与免疫细胞的相关性。从batmanTCM和TCMSP平台获得过敏康组分中所含活性成分与相关靶点,建立“过敏康-活性成分-过敏体质相关靶点”网络并进行调体机制探索。结果 过敏体质人群相较于正常人群存在360个DEGs,共得到816个过敏体质相关基因(相关系数为-0.47,P<0.001),并集中于上皮细胞功能障碍与补体和凝血级联过程。机器学习得到3个基因可作为过敏体质的诊断标记物,在外部验证中表现出稳定的预测能力,其中以SERPINB2的诊断效能最为稳定。5种免疫细胞在过敏体质中具有差异浸润,其中激活的树突状细胞与肥大细胞浸润上调,差异最为显著(P<0.001),SERPINB2与过敏过程中关键免疫细胞密切相关。过敏康调节过敏体质的核心靶点包括ALDH7A1、EDN1、TGFβ1、TP53、AR等,分子机制主要集中于调节组氨酸代谢、细胞表面受体信号、丝裂原活化蛋白激酶(MAPK)信号通路与过氧化物酶体增殖物激活受体(PPAR)信号通路,关键活性成分(灵芝酸DM、5-O-甲基维斯阿米醇苷)与信号通路的核心靶点(MAPK13、PPARγ)均具有显著结合。结论 SERPINB2为主要的过敏体质诊断标记物,激活的树突状细胞与肥大细胞浸润上调是过敏体质的免疫特征,靶向MAPK与PPAR信号通路以调节免疫细胞通讯是过敏康调体干预过敏体质的潜在机制。更多还原

【Abstract】 Objective To identify diagnostic markers and key immune cells for allergic constitution using machine learning methods,and to explore the mechanism of " Allergy Kang " in regulating allergic constitution.Methods Differentially expressed genes(DEGs) in individuals with allergic constitution—comprising patients with various allergic diseases from the Gene Expression Omnibus(GEO) database—were analyzed compared to normal individuals. Weighted Gene Co-expression Network Analysis(WGCNA) was used to identify genes associated with allergic constitution. Proteinprotein interaction(PPI) analysis,Gene Set Enrichment Analysis(GSEA),and Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses were performed. Machine learning algorithms(LASSO regression and SVM-RFE) were employed to identify diagnostic markers for allergic constitution. The CIBERSORT algorithm was used to assess the infiltration levels of 22 types of immune cells,and the correlation between diagnostic markers and immune cells was analyzed. Active components and related targets of " Allergy Kang" were obtained from the batman-TCM and TCMSP platforms,and an " Allergy Kang-Active Components-Allergic Constitution Targets" network was constructed to explore the mechanism of constitution regulation.Results A total of 360 DEGs were identified in individuals with allergic constitution compared to normal individuals. WGCNA revealed 816 genes significantly associated with allergic constitution(correlation coefficient =-0. 47,P<0. 001),which were enriched in epithelial cell dysfunction and complement and coagulation cascades. Machine learning identified three genes as diagnostic markers for allergic constitution,with SERPINB2 demonstrating the most stable diagnostic performance in external validation. Five immune cells showed differential infiltration in allergic constitution,with activated dendritic cells and mast cells being significantly upregulated(P<0. 001). SERPINB2 was closely correlated with key immune cells involved in allergic processes. Core targets of " Allergy Kang" in regulating allergic constitution included ALDH7A1,EDN1,TGFβ1,TP53,and AR. The molecular mechanisms primarily involved the regulation of histidine metabolism,cell surface receptor signaling,MAPK signaling pathways,and PPAR signaling pathways. Key active components(such as ganoderic acid DM and 5-O-methylvisammioside) showed significant binding to core targets of these signaling pathways(e. g.,MAPK13,PPARγ).Conclusion SERPINB2 is a key diagnostic marker for allergic constitution. Upregulated infiltration of activated dendritic cells and mast cells represents an immune characteristic of allergic constitution. Targeting MAPK and PPAR signaling pathways to modulate immune cell communication is a potential mechanism by which " Allergy Kang" regulates allergic constitution.更多还原