Salvianolic Acid B Inhibits Ferroptosis Through ACSL4 Pathway Regulation in sepsis-associated encephalopathy Mice

Sepsis frequently leads to multi-organ injury, with the highly metabolically active nervous system being particularly vulnerable, and ferroptosis has been implicated in driving disease progression. Although Salvianolic acid B (SalB), the most abundant water-soluble active component of Salvia miltiorrhiza Bunge., has demonstrated antioxidant and anti-inflammatory properties, its specific mechanisms in sepsis-associated hippocampal injury remain unclear. To investigate SalB's therapeutic potential against sepsis and its role in mitigating neural damage via ACSL4-mediated ferroptosis, a murine sepsis model was established by cecal ligation and puncture (CLP). SalB's efficacy was evaluated using 7-day survival rates, multi-organ biochemical markers, and critical treatment windows. Inflammatory cytokines were measured by ELISA, and hippocampal morphology was examined histologically. Mechanistic studies included Fe²⁺ staining and lipid peroxidation assays, while protein arrays and Western blotting clarified SalB's interaction with ACSL4, confirming its anti-ferroptotic role. Our results show that SalB significantly improved survival in CLP-induced septic mice, reduced levels of inflammatory factors, alleviated hippocampal neuronal damage, and preserved blood-brain barrier integrity. Data from biochemical assays and Western blot analysis indicated that SalB suppresses ferroptosis by modulating the ACSL4/GPX4 pathway, supporting its therapeutic role in septic hippocampal injury. Additionally, protein array and molecular docking studies provided evidence that SalB likely exerts its pharmacological activity by competitively inhibiting the substrate CoA binding to ACSL4 at amino acid residues LYS-572, LEU-574, and SER-607. In conclusion, SalB protects against sepsis-induced hippocampal injury by targeting ACSL4-mediated ferroptosis, offering a novel herbal-based strategic direction for sepsis treatment.