Melittin ameliorates inflammation in mouse acute liver failure via inhibition of PKM2-mediated Warburg effect
Melittin ameliorates inflammation in mouse acute liver failure via inhibition of PKM2-mediated Warburg effect
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Melittin 通过抑制 PKM2 介导的 Warburg 效应改善小鼠急性肝衰竭的炎症
DOI：
10.1038/s41401-020-00516-0
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发表时间：
2020-09-16
影响因子：
8.2
通讯作者：
Li, Ning
中科院分区：
文献类型：
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作者：
Fan, Xue-gong;Pei, Si-ya;Li, Ning
急性肝衰竭（ALF）是一种致命的临床综合征，无特效药物。最近的证据表明，调节巨噬细胞抑制炎症可能是 ALF 治疗的一种有前景的策略。在这项研究中，我们研究了蜂毒肽（蜂毒的主要肽成分）在 ALF 小鼠模型和体外 LPS 刺激的巨噬细胞中的潜在治疗作用，并阐明了潜在的机制。通过腹腔注射d-半乳糖胺/LPS在小鼠中诱导ALF。然后用蜂毒肽（2、4和8mg/kg，腹膜内注射）治疗小鼠。我们发现，蜂毒肽治疗显着提高了死亡率，减轻了严重症状和体征，并减轻了 ind-半乳糖胺/LPS 诱导的 ALF 小鼠的肝脏炎症，最佳剂量为 4mg/kg。此外，蜂毒肽在有效剂量内并没有引起明显的体内毒性。在 LPS 刺激的 RAW264.7 巨噬细胞中，蜂毒肽 (0.7μM) 发挥抗氧化和抗炎作用。我们发现 LPS 刺激通过增加糖酵解速率促进巨噬细胞的有氧糖酵解，上调 Warburg 效应相关酶和代谢物（包括乳酸、LDHA、LDH 和 GLUT-1）的水平，并激活 Akt/mTOR/PKM2/HIF-1α 信号传导。蜂毒肽治疗可抑制丙酮酸激酶 (PKM2) 的 M2 亚型，从而破坏 Warburg 效应以减轻炎症。分子对接分析证实蜂毒肽靶向 PKM2。在 LPS 刺激的 RAW264.7 巨噬细胞中，敲除 PKM2 会产生与蜂毒肽类似的抗炎作用。 Ind-半乳糖胺/LPS诱导的ALF小鼠，蜂毒肽治疗显着降低肝脏中PKM2和HIF-1α的表达水平。这项工作表明，蜂毒肽通过靶向 PKM2 抑制有氧糖酵解，从而抑制巨噬细胞活化介导的炎症，这凸显了使用蜂毒肽治疗 ALF 的新策略。
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 ofd-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 ind-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. Ind-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.