Zishuo Cheng ^1,2 Shun Huang ^1,2 Qiya Tang ^1,2 Danlan Zhang ^1,2 Lan Huang ^1,2*

• ¹ Chongqing Medical University, Chongqing, China
• ² Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Stomatological Hospital of Chongqing Medical University, Chongqing, China

Objective: Mitochondrial metabolic reprogramming in macrophages is crucial in the development and progression of inflammation. Given vitamin A's antioxidant properties and its therapeutic effects on inflammation, this study aims to elucidate how vitamin A influences mitochondrial metabolic reprogramming in inflammatory states, specifically in periodontitis, through genetic bioinformatics and experimental methods.Method: The study utilized the GSE16134 dataset from the Gene Expression Omnibus (GEO) database, focusing on human periodontitis. Vitamin A-targeted genes (ATGs) were identified and analyzed using CIBERSORT to explore their role in inflammation. Cluster analysis revealed two phenotypes associated with ATGs, showing differential expression of genes like COX1, IL-1β, and STAT3, and immune activation patterns. Weighted Gene Co-expression Network Analysis (WGCNA) identified 145 markers correlated with ATG-guided phenotypes and inflammation. Machine learning models, combined with Gene Set Variation Analysis (GSVA), identified five key genes (RGS1, ACAT2, KDR, TUBB2A, TDO2) linked to periodontitis. Cell Type-Specific Enrichment Analysis (CSEA) highlighted macrophages as critical in metabolic reprogramming, validated by external datasets with an AUC of 0.856 in GSE10334 and 0.750 in GSE1730678856. Experimental validation showed vitamin A's role in suppressing endoplasmic reticulum stress and altering mitochondrial dynamics, as well as metabolic reprogramming influencing inflammation via the STAT3 pathway in RAW 264.7 cells.Results: The study identified 13 differentially expressed ATGs in periodontitis, showing strong correlations with inflammation, particularly in plasma cells, macrophages, dendritic cells, neutrophils, and mast cells. Two ATG-guided phenotypes were identified, differing in gene expression and immune activation. WGCNA and machine learning models identified 145 markers and five key genes associated with periodontitis. GSVA and CSEA analyses highlighted the JAK- STAT pathway and macrophage involvement in metabolic reprogramming. Experimental data confirmed vitamin A's effects on mitochondrial dynamics and metabolic reprogramming through the STAT3 pathway.The study demonstrates that vitamin A's therapeutic effect on periodontitis is mediated through JAK-STAT pathway-guided mitochondrial metabolic reprogramming in macrophages. It identifies two genetic and immune-related phenotypes and five genetic identifiers associated with periodontitis risk.