Potential Mechanisms of Acupuncture Treatment for Rheumatoid Arthritis: A Study Based on Network Topology and Machine Learning

Objective: Rheumatoid arthritis (RA) is a systemic autoimmune disease requiring multi-target therapeutic strategies. Acupuncture, a holistic therapy in traditional Chinese medicine (TCM), has shown clinical efficacy in RA, yet its molecular mechanisms remain elusive. By integrating network topology and machine learning methods, decode the systemic regulatory effects of acupuncture on RA of acupuncture on RA. Methods: Data on the interactions between acupuncture-affected endogenous compounds and RA-related targets were extracted from databases, built a multi-dimensional interaction network to map the interactions between acupuncture and RA. screened RA differentially expressed genes (DEGs) from GEO, intersecting with acupuncture-responsive genes. Utilize clusterProfiler for KEGG/GO enrichment analysis of these DEGs, Analyze the immune microenvironment using CIBERSORTx and xCell algorithms. Utilize ConsensusClusterPlus (R package) for unsupervised clustering to obtain DEGs. Subsequently, identify key genes using an ensemble machine learning model (GLM/SVM/XGB/RF) and create nomograms. Apply TwoSample MR and colocalization analysis to validate the causal relationship between core acupuncture-affected DEGs and RA risk. Results: This study identified 10 acupuncture-regulated metabolites and 49 RA-related DEGs. KEGG analysis showed DEGs enriched in immune pathways including the JAK/STAT pathway mediating inflammatory responses, the T-cell receptor signaling pathway involved in T cell differentiation and the TNF signaling pathway. Immunome profiling based on the CIBERSORT algorithm indicated that DEGs were primarily enriched in key immune cell subpopulations such as M1 macrophages, activated CD4⁺ T cells, Tregs, and B lymphocytes. Machine learning identified five key genes associated with immune infiltration (STAT1, GAPDH, JAK2, PTGS2, MDM2). MR/colocalization confirmed acupuncture-regulated STAT1 expression positively correlated with RA genetic susceptibility, highlighting STAT1-mediated JAK/STAT pathway in immune remodeling. Conclusion: STAT1, GAPDH, JAK2, PTGS2, and MDM2 may be potential targets for acupuncture treatment of RA. Acupuncture may achieve systemic immune regulation by synergistically targeting multiple pathways (JAK/STAT, TNF) and immune cells (M1 macrophages, CD4⁺ T cells), This initiative integrates the holistic philosophy of TCM with the precision of AI-driven medical science.