Identification and multi-layered experimental validation of feature genes related to butyrate metabolism and oxidative stress in sepsis

The mechanisms underlying sepsis remain poorly understood and specific therapeutic options remain lacking. Accumulating evidence suggests that butyrate metabolism and oxidative stress (OS) are involved in the pathogenesis of sepsis. This study aims to elucidate molecular characteristics of butyrate metabolism and OS-related genes in sepsis and find potential therapeutic targets. Using sepsis-related datasets, 876 differential expression genes (DEGs) were screened and intersected with butyrate metabolism and OS-related genes to yield 8 differentially expressed butyrate metabolism and OS-related genes (DEBORGs). Machine learning algorithms finally identified 3 feature genes (ALDH1A1, CYP1B1, and GADD45A) with favourable diagnostic values for sepsis. CIBERSORT immune infiltration analysis demonstrated strong associations between feature genes and the infiltration proportions of various immune cell types, such as monocyte, macrophage, and neutrophil. Cellular experiment and prospectively collected clinical samples were utilized to validate the expression levels and diagnostic efficacy of feature genes. The scRNA-seq analysis indicated that monocyte plays a crucial role in mediating the effect of feature genes on sepsis. This study elucidates that ALDH1A1, CYP1B1, and GADD45A are feature genes linked to butyrate metabolism and OS in sepsis, with monocytes being the primary cells involved, providing novel insights into the sepsis pathogenesis and potential therapeutic targets.