Nur77 是核受体超家族的孤儿成员，可调节炎症性疾病，是治疗炎症的治疗靶点。当归中的苯酞具有抗炎活性。本研究旨在筛选当归提取物中能够通过靶向Nur77发挥抗炎活性的化合物。为更好地阐明中药靶向线粒体以实现多种临床疗效提供新的理论支持。采用蛋白质印迹法评估了苯酞在肿瘤坏死因子-α（TNF-α）刺激的HepG2细胞中的抗炎能力。通过分子对接、表面等离子体共振和细胞热位移测定验证了苯酞和 Nur77 之间的相互作用。进行免疫共沉淀、蛋白质印迹和免疫染色以确定分子机制。在脂多糖（LPS）/d-半乳糖胺（d-GalN）诱导的急性肝炎和肝损伤小鼠急性肝炎和肝损伤模型中评估了苯酞的体内抗炎活性。最后，在斑马鱼实验中评估了苯酞的毒性。在从中华黄芪中分离的27种苯酞化合物中，tokinolide B (TB)表现出最好的Nur77结合能力和最好的抗炎活性，并且不诱导细胞凋亡。体内和体外实验表明，TB促进Nur77从细胞核易位到线粒体，并与肿瘤坏死因子受体相关因子2（TRAF2）和隔离体1（p62）相互作用，诱导线粒体自噬，从而发挥抗炎功能。 TB 显着抑制 LPS/d-GalN 诱导的小鼠急性肝炎和肝损伤。在斑马鱼实验中，TB 还表现出明显低于雷公藤红素的毒性。这些发现表明，TB 通过促进 Nur77 与 TRAF2 和 p62 相互作用，从而诱导线粒体自噬来抑制炎症。这些发现为开发新型抗炎药物提供了有希望的方向，增强了对苯酞化合物的了解，并突出了传统中草药的治疗潜力。版权所有 © 2024 Elsevier GmbH。版权所有。

Nur77, an orphan member of the nuclear receptor superfamily, regulates inflammatory diseases and is a therapeutic target for treating inflammation. Phthalides in Angelica sinensis exhibit anti-inflammatory activity.This study aimed to screen compounds from A. sinensis phthalide extract that could exert anti-inflammatory activity by targeting Nur77. To provide new theoretical support for better elucidation of Chinese medicine targeting mitochondria to achieve multiple clinical efficacies.The anti-inflammatory capacity of phthalides was assessed in tumor necrosis factor-alpha (TNF-α)-stimulated HepG2 cells using western blotting. The interaction between phthalides and Nur77 was verified by molecular docking, surface plasmon resonance, and cellular thermal shift assay. Co-immunoprecipitation, western blotting, and immunostaining were performed to determine the molecular mechanisms. The in vivo anti-inflammatory activity of the phthalides was evaluated in a lipopolysaccharide (LPS)/d-galactosamine (d-GalN)-induced acute hepatitis and liver injury mouse model of acute hepatitis and liver injury. Finally, the toxicity of phthalide toxicity was assessed in zebrafish experiments.Among the 27 phthalide compounds isolated from A. sinensis, tokinolide B (TB) showed the best Nur77 binding capacity and, the best anti-inflammatory activity, which was induced without apoptosis. In vivo and in vitro experiments showed that TB promoted Nur77 translocation from the nucleus to the mitochondria and interacted with tumor necrosis factor receptor-associated factor 2 (TRAF2) and sequestosome 1 (p62) to induce mitophagy for anti-inflammatory functions. TB substantially inhibited LPS/d-GalN-induced acute hepatitis and liver injury in mice. TB also exhibited significantly lower toxicity than celastrol in zebrafish experiments.These findings suggested that TB inhibits inflammation by promoting Nur77 interaction with TRAF2 and p62, thereby inducing mitophagy. These findings offer promising directions for developing novel anti-inflammatory agents, enhance the understanding of phthalide compounds, and highlight the therapeutic potential of traditional Chinese herbs.Copyright © 2024 Elsevier GmbH. All rights reserved.