Farrerol alleviates hepatic ischemia-reperfusion injury by regulating the CD14/MyD88/NF-κB signaling pathway

Hepatic ischemia-reperfusion injury (HIRI) remains a critical complication in liver surgery with limited pharmacological options, and while Farrerol (FA), a natural dihydroflavone, has shown protective effects in other organ IR models, its role in HIRI has never been explored. In this study, we established a murine model of 70% HIRI models and found that intravenous administration of FA (10 and 20 mg/kg) at the onset of reperfusion significantly attenuated HIRI, as evidenced by reduced serum ALT/AST levels, improved histopathology, decreased hepatocyte apoptosis, mitigated oxidative stress, and downregulated inflammatory cytokine expression. To uncover the underlying mechanism, we performed transcriptomic analysis of liver tissues, which unexpectedly identified CD14 as a key target and revealed that FA markedly suppressed the MyD88/NF-κB signaling pathway. Mechanistic validation in LPS-stimulated RAW264.7 macrophages confirmed that FA dose-dependently inhibited M1 polarization and inflammatory responses. Crucially, CD14 overexpression in macrophages largely abrogated the anti‑inflammatory effects of FA. By contrast, CD14 silencing mimicked FA‑mediated suppression of NF‑κB activation and cytokine release, and FA treatment exerted no further inhibitory effects in CD14‑silenced macrophages. Collectively, our findings demonstrate for the first time that FA alleviates HIRI by downregulating macrophage CD14 expression, thereby inhibiting the downstream MyD88/NF-κB pathway and its associated inflammatory cascade, positioning FA as a promising therapeutic candidate and highlighting CD14 as a viable target for HIRI intervention.