Abstract
BACKGROUND: Type 2 diabetes mellitus (T2DM) is linked to elevated blood glucose due to insulin resistance. Empagliflozin has demonstrated efficacy in T2DM management, with potential immune-modulatory effects. This study aimed to investigate the immune-related mechanisms of empagliflozin in T2DM treatment. METHODS: Gene expression profiles were obtained from common databases, and immune-related differentially expressed genes (IRDEGs) were identified using the "limma" R package. The STRING database and Cytoscape software were utilized to construct a protein-protein interaction network and identify key IRDEGs. Molecular docking was performed to predict the interaction between empagliflozin and PIK3CA. Pathways related to PIK3CA were explored using GSEA, and relationship of PIK3CA with immune cells was analyzed using single-cell RNA sequencing analysis. The effects of empagliflozin on high glucose-induced RAW264.7 macrophages and PI3K/AKT signaling were assessed using CCK-8, fluorescence detection, qRT-PCR, and Western blotting. RESULTS: We identified 109 IRDEGs in T2DM, with PIK3CA as a key gene. Empagliflozin showed binding affinity to PIK3CA, which was linked to immune cell interactions and inflammatory responses. Single-cell RNA sequencing analysis revealed the interaction of PIK3CA with macrophages. In high glucose-induced RAW264.7 macrophages, PIK3CA expression was elevated. Empagliflozin ameliorated the high glucose-induced cell injury and inhibited the expression of PIK3CA in macrophages. Additionally, empagliflozin treatment reduced the expression of CD44 and ITGAV, IL-6, and TNF-α, and increased the p-PI3K/PI3K ratio. CONCLUSION: Empagliflozin's therapeutic effects in T2DM may be mediated through the modulation of immune pathways, particularly by targeting PIK3CA within the PI3K/AKT signaling pathway.