Acupuncture alleviates chronic airway inflammation in obese asthmatic mice by downregulating Vnn1 and FAM126B

doi:10.12122/j.issn.1673-4254.2025.12.05

Abstract

Abstract:

Objective To identify the differentially expressed genes in obese asthma versus non-obese asthma and evaluate the effect of acupuncture on chronic airway inflammation in obese mice with asthma. Methods The key genes of obesity-related asthma were screened using GSE110551 dataset from the GEO database, and the characteristic genes were selected from the genes with the highest correlation with T cells using Lasso regression and SVM feature selection algorithms. Fifty C57BL/6J mice (5-6 weeks old) were randomized equally into 5 groups, including a normal feeding (control) group, a high-fat feeding group, and 3 high-fat feeding and ovalbumin sensitization groups with intraperitoneal injections with saline or dexamethasone (DEX), or treated with acupuncture. Western blotting, qPCR, and flow cytometry were used to analyze the changes in the expressions of the key genes and inflammation in the airway of the mice. Results FAM126B and VNN1 were identified as the characteristic genes in obesity-related asthma for subsequent analysis. The mice with high-fat feeding and ovalbumin sensitization showed the highest expression levels of Vnn1 and FAM126B among the 5 groups, with also significantly decreased Treg cell percentage and obvious inflammatory cell infiltration in the lungs. Treatment with DEX and acupuncture both significantly decreased the number of infiltrating inflammatory cells and increased the percentage of Treg cells in airway of the mouse models of obesity-related asthma. HIF-1α was identified as a key regulatory factor for asthmatic inflammation, and its expression level was significantly increased in the asthmatic mouse models but obviously lowered after acupuncture treatment or dexamethasone therapy. Conclusion Vnn1 and FAM126B may serve as the key therapeutic targets for treatment of obese asthma patients. Acupuncture treatment may downregulate airway HIF-1α by reducing the expressions of Vnn1 and FAM126B and increasing the number of Treg cells.

Key words: obesity, asthma, Vnn1, FAM126B, acupuncture

Xiaofeng LI, Taochun YE, Lu XI, Chunqiao LI, Huihui LIU. Acupuncture alleviates chronic airway inflammation in obese asthmatic mice by downregulating Vnn1 and FAM126B[J]. Journal of Southern Medical University, 2025, 45(12): 2573-2584.

Figures/Tables 8

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.

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.

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.

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.

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.

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.

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.

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.

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