Formononetin targets TLR4/MyD88 signaling to attenuate Escherichia coli-induced sepsis: Integration of in vivo, network pharmacology and experimental validation.

Abstract

Sepsis, a life-threatening systemic inflammatory syndrome caused by dysregulated host responses to infection, frequently originates from Gram-negative pathogens such as Escherichia coli (E. coli). While antibiotics remain the mainstay treatment, rising antimicrobial resistance necessitates alternative therapeutic strategies. Formononetin (FMN), a bioactive isoflavone derived from traditional Chinese medicine, exhibit potent anti-inflammatory and antioxidant properties. This study employed an integrative approach combining in vivo pharmacodynamic evaluation, network pharmacology, and experimental validation to systematically investigate FMN's therapeutic potential against E. coli induced sepsis. In vivo studies in a murine sepsis model showed FMN significantly reduced E. coli sepsis induced body weight loss, clinical manifestations, histopathological changes, and suppressed proinflammatory cytokines (e.g. IL-6 and TNF-α). Network pharmacology identified 28 putative targets interacted with the E. coli sepsis-related site and molecular docking suggested possible interactions between FMN and key inflammatory mediators such as Toll-like receptor 4 (TLR4) and tumor necrosis factor (TNF). qRT-PCR indicated that FMN modulated the transcription of TLR4/MyD88-related genes, with downstream reductions in the mRNA levels of TLR4, MyD88, iNOS and IL-6. These findings suggest that FMN may be a promising candidate for further investigation as a multi-target modulator in sepsis.