A Systematic Exploration of Gut Microbiota–Driven Blood Metabolites in Sepsis: An Integrated Bioinformatics and Genetic Association Study

Yanhuo Zhang¹Jun Liu¹Yaling Peng²Xingyu Li^3*Peng Zhu^1*

• ¹Department of Gastrointestinal Anorectal Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China
• ²The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi, China
• ³The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China

Alterations in the blood metabolome are closely associated with sepsis, while the gut microbiota (GM) plays a crucial role in modulating both sepsis progression and circulating metabolites. To determine whether the effects of the GM on sepsis are mediated through blood metabolites, we performed a two-sample Mendelian randomization (MR) analysis combined with a two-step MR framework to identify potential metabolic mediators. Comprehensive bioinformatics analyses were integrated to construct interaction networks using Cytoscape, and pharmacodynamic experiments were conducted in a murine sepsis model. We identified 23 GM taxa and 169 blood metabolites significantly associated with sepsis. Two-step MR analysis revealed that 15 metabolites mediated the causal relationships between 12 GM taxa and sepsis, with mediation proportions ranging from 3.70% to 13.70%. A total of 131 potential molecular targets were predicted for these metabolites, and network analysis highlighted five key metabolites and seven central targets. Molecular docking demonstrated strong binding affinities between these metabolites and their targets. Notably, gulonic acid (GA) and 4-hydroxyphenylacetic acid (4-HPA), driven by Lentisphaerae, Lentisphaeria, and Victivallales, significantly improved survival and attenuated organ injury and inflammation in septic mice. Collectively, this study provides evidence supporting a causal role of the gut microbiota in sepsis, mediated in part by blood metabolites. These findings highlight the therapeutic potential of targeting both the gut microbiota and GM-driven metabolites as novel interventions for sepsis.