Abstract
Acute liver failure (ALF) is a fatal clinical syndrome with no special drug. Recent evidence shows that modulation of macrophage to inhibit inflammation may be a promising strategy for ALF treatment. In this study we investigated the potential therapeutic effects of melittin, a major peptide component of bee venom both in mice model of ALF and in LPS-stimulated macrophages in vitro, and elucidated the underlying mechanisms. ALF was induced in mice by intraperitoneal injection of d-galactosamine/LPS. Then the mice were treated with melittin (2, 4, and 8 mg/kg, ip). We showed that melittin treatment markedly improved mortality, attenuated severe symptoms and signs, and alleviated hepatic inflammation in d-galactosamine/LPS-induced ALF mice with the optimal dose being 4 mg/kg. In addition, melittin within the effective doses did not cause significant in vivo toxicity. In LPS-stimulated RAW264.7 macrophages, melittin (0.7 μM) exerted anti-oxidation and anti-inflammation effects. We showed that LPS stimulation promoted aerobic glycolysis of macrophages through increasing glycolytic rate, upregulated the levels of Warburg effect-related enzymes and metabolites including lactate, LDHA, LDH, and GLUT-1, and activated Akt/mTOR/PKM2/HIF-1α signaling. Melittin treatment suppressed M2 isoform of pyruvate kinase (PKM2), thus disrupted the Warburg effect to alleviate inflammation. Molecular docking analysis confirmed that melittin targeted PKM2. In LPS-stimulated RAW264.7 macrophages, knockdown of PKM2 caused similar anti-inflammation effects as melittin did. In d-galactosamine/LPS-induced ALF mice, melittin treatment markedly decreased the expression levels of PKM2 and HIF-1α in liver. This work demonstrates that melittin inhibits macrophage activation-mediated inflammation via inhibition of aerobic glycolysis by targeting PKM2, which highlights a novel strategy of using melittin for ALF treatment.
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Acknowledgements
This work was supported by the project funded by China Postdoctoral Science Foundation (no’s. 2016M600639 and 2017T100614), the National Natural Science Foundation of China (no’s. 81673719, 81970550, 81700561 and 81873574), and Natural Science Foundation of Hunan Province (no’s. 2019JJ30042 and 11JJ6072).
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NL performed animal experiments and wrote the manuscript; DZ, SYP, and PCZ performed the cell experiments; XWH and TL performed the molecular docking analysis; ZBH and YH wrote part of the manuscript; XGF designed the experiments and edited the manuscript; YW and XGF designed the experiments, analyzed the data, XGF and SYP revised all figures, and revised the manuscript.
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Fan, Xg., Pei, Sy., Zhou, D. et al. Melittin ameliorates inflammation in mouse acute liver failure via inhibition of PKM2-mediated Warburg effect. Acta Pharmacol Sin 42, 1256–1266 (2021). https://doi.org/10.1038/s41401-020-00516-0
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DOI: https://doi.org/10.1038/s41401-020-00516-0
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