图1 差异表达基因(DEGs)的筛选:免疫浸润和WGCNA

Fig.1 Analysis of the differentially expressed genes (DEGs): Immune Infiltration and WGCNA. A: Stacked bar graph showing immune infiltration of multiple samples from GSE110551. B: Volcano plots showing the differences in gene expressions. C: Heat map showing the connections among different immune cell types. D: Selection of a soft threshold in a weighted gene co-expression network analysis (WGCNA). E: Results of hierarchical clustering based on gene expression data. F: Heat map showing connections of different modules and the specific features of each cell.

图2 差异表达基因(DEGs)的筛选:GO/KEGG富集分析和机器学习

Fig.2 Screening of the DEGs using GO/KEGG enrichment analysis and machine learning. A,B: Results of pathway analysis of genes in the green module using the Metascape website. C: Lasso regression curve diagram. D: Selection of the optimal λ value to balance model complexity and predictive performance. E: Evaluation of feature genes in obesity-asthma using the Support Vector Machine Recursive Feature Elimination (SVM-RFE) algorithm. F: Venn diagram of LASSO and SVM-RFE: two intersecting genes, FAM126B and VNN1, were obtained.

图3 FAM126B和VNN1涉及的免疫因子、GSVA分析

Fig.3 Immunological factors associated with FAM126B and VNN1 and GSVA analysis. A-E: Bubble plots showing the correlation between different chemokines, immunosuppressive factors, immunostimulatory factors, MHC, chemokine receptors, and the two specific genes (FAM126B and VNN1). F-G: GSVA bar charts for FAM126B and VNN1. Patients with high FAM126B and VNN1 expressions exhibit enrichment in different signaling pathways.

图4 FAM126B和VNN1的GSEA分析、多个转录因子共同调控富集分析以及mRNA-miRNA关系

Fig.4 GSEA analysis of FAM126B and VNN1, co-regulation enrichment analysis of multiple transcription factors, and mRNA-miRNA relationships. A,B: GSEA results and network diagram of FAM126B, enriched in the IL-17 signaling pathway, inositol phosphate metabolism, and TNF signaling pathway. C, D: GSEA results and network diagram for VNN1, enriched in the p53 signaling pathway, pyrimidine metabolism, and TNF signaling pathway. E: Enrichment analysis and motif-TF annotation of transcription factors. F: miRNA network diagram for FAM1268 and VNN1 using Cytoscape.

图5 肥胖哮喘小鼠模型及小鼠体质量随时间的变化

Fig.5 Obese asthma modeling in mice and weekly changes of the related parameters of the mice. A: Flow chart of animal experiment. B: Location of mice acupoints. C: Line chart of body weight changes of the mice. D: Line chart of lee's index in mice. E: Line chart of RF test of the mice. △P<0.05 vs COH. CON: Normal feeding group; FA: High-fat diet feeding group; ACU: High-fat feeding group with OVA sensitization and treatment with acupuncture; DEX: High-fat feeding group with OVA sensitization and 2 mg/kg dexamethasone gavage; FAA: High-fat feeding and OVA sensitization group.

图6 动物实验HE染色和血常规结果

Fig.6 HE staining of the lung tissues and blood routine results in each group. A-F: Pathological inflammatory cell infiltration in mice, including lymphocytes, eosinophils, and neutrophils (×100). The inflammation score was 0 in CON group, 1 in FA group, 3 in ACU group, 4 in DEX group, and 5 in FAA group. G: Mouse blood routine test results for neutrophils, lymphocytes, monocyte and eosinophils. △P<0.05 vs CON group; ○P<0.05 vs FA group; &P<0.05 vs ACU group; *P<0.05 vs DXE group.

图7 针刺与地塞米松治疗效果

Fig.7 Therapeutic effects of acupuncture and dexamethasone in the mouse models. A: ELISA detection results of bronchoalveolar lavage fluid for IL-4, IL-5, IL-17, IL-1β, and TNF-α. B, C: Western blotting for detecting IL-1β expressions in each group. D-F: Flow cytometry for analysis of Treg cells. △P<0.05 vs CON group; ○P<0.05 vs FA group; &P<0.05 vs ACU group; *P<0.05 vs DXE group.

图8 HIF-1α、FAM126B和Vnn1在肥胖哮喘中的表达情况

Fig.8 Expression of HIF-1α, FAM126B and Vnn1 in mice with obese asthma. A-C: Immunofluorescence staining of Vnn1, FAM126B, and HIF-1α in the lung tissues (×100). D: Bar charts showing mRNA expression levels of HIF-1α, Vnn-1 and FAM126B in the lung tissues. △P<0.05 vs CON group; ○P<0.05 vs FA group; &P<0.05 vs ACU group; *P<0.05 vs DXE group.