Abstract：
PANoptosis is manifested with simultaneous activation of biomarkers for both pyroptotic, apoptotic and necroptotic signaling via the molecular platform PANoptosome and it is involved in pathologies of various inflammatory diseases including hemophagocytic lymphohistiocytosis (HLH). Scutellarin is a flavonoid isolated from herbal Erigeron breviscapus (Vant.) Hand.-Mazz. and has been shown to possess multiple pharmacological effects, but it is unknown whether scutellarin has any effects on PANoptosis and related inflammatory diseases. In this study, we found that scutellarin inhibited cell death in bone marrow-derived macrophages (BMDMs) and J774A.1 cells treated with TGF-β-activated kinase 1 (TAK1) inhibitor 5Z-7-oxozeaenol (OXO) plus lipopolysaccharide (LPS), which has been commonly used to induce PANoptosis. Western blotting showed that scutellarin dose-dependently inhibited the activation biomarkers for pyroptotic (Caspase-1p10 and GSDMD-NT), apoptotic (cleaved Casp3/8/9 and GSDME-NT), and necroptotic (phosphorylated MLKL) signaling. The inhibitory effect of scutellarin was unaffected by NLRP3 or Caspase-1 deletion. Interestingly, scutellarin blocked the assembly of PANoptosome that encompasses ASC, RIPK3, Caspase-8 and ZBP1, suggesting its action on upstream signaling. Consistent with this, scutellarin inhibited mitochondrial damage and mitochondrial reactive oxygen species (mtROS) generation in cells treated with OXO+LPS. Further, mito-TEMPO that can scavenge mtROS significantly inhibited OXO+LPS-induced PANoptotic cell death. In line with the in vitro results, scutellarin markedly alleviated systemic inflammation, multiple organ injury, and activation of PANoptotic biomarkers in mice with HLH. Collectively, our data suggest that scutellarin can inhibit PANoptosis by suppressing mitochondrial damage and mtROS generation and thereby mitigating multiple organ injury in mice with inflammatory disorders.
摘要：
PANoptosis表现为同时激活生物标志物，通过分子平台PANoptosome的凋亡和坏死信号传导，它参与各种炎性疾病的病理，包括噬血细胞性淋巴组织细胞增多症（HLH）。Scutellarin是从草药灯盏花（Vant。）手。-爵士.并已被证明具有多种药理作用，但目前尚不清楚灯盏乙素是否对PANoptosis和相关的炎症性疾病有影响。在这项研究中,我们发现灯盏乙素抑制了用TGF-β激活的激酶1（TAK1）抑制剂5Z-7-氧代玉米烯醇（OXO）加脂多糖（LPS）处理的骨髓源性巨噬细胞（BMDMs）和J774A.1细胞的细胞死亡，通常用于诱导PANoptosis。Westernblotting显示，灯盏乙素剂量依赖性地抑制了热变性的激活生物标志物（Caspase-1p10和GSDMD-NT），凋亡（裂解的Casp3/8/9和GSDME-NT），和坏死（磷酸化MLKL）信号。灯盏乙素的抑制作用不受NLRP3或Caspase-1缺失的影响。有趣的是,scutellarin阻止了包含ASC的PANoptosome的组装，RIPK3，Caspase-8和ZBP1，表明其对上游信号传导的作用。与此一致，灯盏乙素抑制OXO+LPS处理细胞的线粒体损伤和线粒体活性氧（mtROS）的生成。Further,可以清除mtROS的mito-TEMPO显着抑制OXOLPS诱导的PANopotic细胞死亡。根据体外结果，灯盏乙素可显着缓解全身炎症，多器官损伤,以及HLH小鼠中PANopottic生物标志物的激活。总的来说，我们的数据表明,灯盏乙素可以通过抑制线粒体损伤和mtROS的生成,从而减轻炎症性疾病小鼠的多器官损伤,从而抑制PANoptosis.
