Targeting hepatocyte-derived LRG1 with dothiortone alleviates hepatic ischemia–reperfusion injury and remote renal damage via suppressing NETosis

Abstract Hepatic ischemia–reperfusion injury (HIRI), a major complication following liver surgery, involves neutrophil infiltration and hepatocyte apoptosis that contribute to graft failure and multiorgan injury; however, the role of neutrophil–hepatocyte crosstalk remains unclear. Through proteomic profiling, leucine-rich α-2 glycoprotein 1 (LRG1) was identified as a markedly upregulated protein in mouse HIRI models, a finding corroborated by elevated plasma LRG1 levels in patients who underwent hepatectomy. In mice, hepatocyte-derived LRG1 precedes neutrophil extracellular trap (NET) formation and hepatocyte apoptosis. Hepatocyte-specific Lrg1 knockout attenuated liver injury, inflammation, neutrophil infiltration, NET formation, and apoptosis. Mechanistically, LRG1 secreted from hepatocytes binds to TGFβ receptor 1 (TGFβR1) on neutrophils, activating SMAD3 to promote Pad4 transcription and NETosis. Subsequent NETs initiate AIM2-dependent hepatocyte apoptosis and propagate injury to the kidneys, resulting in AIM2-mediated renal tubular damage—effects ameliorated by DNase I treatment or AIM2 deficiency. Importantly, molecular docking and surface plasmon resonance analyses identified dothiortone, a mangrove-derived polyketide, as a high-affinity LRG1 inhibitor. Dothiortone effectively suppressed NETosis and mitigated HIRI-induced hepatic and renal injury in vivo. These findings demonstrate that hepatocyte-derived LRG1 exacerbates HIRI and remote kidney injury via the TGFβR1–SMAD3–PAD4 axis and NET-driven AIM2-dependent apoptosis, highlighting LRG1 inhibition by dothiortone as a promising therapeutic strategy for HIRI and its systemic complications.