脑胶质瘤是最常见的原发性恶性脑肿瘤，生存率低且治疗选择有限。自然苯并芘生物碱黄连碱能够在多种癌细胞中表现出抗肿瘤作用，但其在脑胶质瘤中的分子靶标和信号转导过程仍不清楚。本研究探究了黄连碱在脑胶质瘤细胞系和小鼠移植瘤模型中的作用机制。结果表明，黄连碱诱导细胞死亡与RIP1/RIP3依赖性的坏死程序有关，而非在早期的凋亡程序。机制研究揭示了坏死过程与线粒体功能障碍之间的相互作用，黄连碱诱导线粒体ROS的产生、线粒体去极化、ATP水平降低和线粒体碎片化，这是RIP1依赖性坏死程序激活的重要触发机制。同时，PINK1和parkin依赖性线粒体自噬促进了CHE处理的脑胶质瘤细胞中受损线粒体的清除，而使用CQ抑制线粒体自噬则有助于选择性促进CHE诱导的坏死程序。此外，CHE引导的胞质钙离子也是诱导线粒体损伤和坏死程序重要的“启动信号”。抑制线粒体ROS有助于打断线粒体受损和RIPK1/RIPK3坏死亡团的正反馈作用。最后，使用CHE抑制U87移植瘤的皮下肿瘤生长并不会导致明显的体重下降和多器官毒性。总之，本研究有助于阐明CHE通过线粒体ROS介导的RIP1-RIP3-Drp1复合物形成来促进Drp1线粒体转位从而诱导坏死程序。研究结果表明，CHE有望成为治疗脑胶质瘤的一种新型治疗策略。版权所有 © 2023 Elsevier Inc. 发表。

Glioma is the most common primary malignant brain tumor with poor survival and limited therapeutic options. Chelerythrine (CHE), a natural benzophenanthridine alkaloid, has been reported to exhibit the anti-tumor effects in a variety of cancer cells. However, the molecular target and the signaling process of CHE in glioma remain elusive. Here we investigated the underlying mechanisms of CHE in glioma cell lines and glioma xenograft mice model. Our results found that CHE-induced cell death is associated with RIP1/RIP3-dependent necroptosis rather than apoptotic cell death in glioma cells at the early time. Mechanism investigation revealed the cross-talking between necroptosis and mitochondria dysfunction that CHE triggered generation of mitochondrial ROS, mitochondrial depolarization, reduction of ATP level and mitochondrial fragmentation, which was the important trigger for RIP1-dependent necroptosis activation. Meanwhile, PINK1 and parkin-dependent mitophagy promoted clearance of impaired mitochondria in CHE-incubated glioma cells, and inhibition of mitophagy with CQ selectively enhanced CHE-induced necroptosis. Furthermore, early cytosolic calcium from the influx of extracellular Ca2+ induced by CHE acted as important "priming signals" for impairment of mitochondrial dysfunction and necroptosis. Suppression of mitochondrial ROS contributed to interrupting positive feedback between mitochondrial damage and RIPK1/RIPK3 necrosome. Lastly, subcutaneous tumor growth in U87 xenograft was suppressed by CHE without significant body weight loss and multi-organ toxicities. In summary, the present study helped to elucidate necroptosis was induced by CHE via mtROS-mediated formation of the RIP1-RIP3-Drp1 complex that promoted Drp1 mitochondrial translocation to enhance necroptosis. Our findings indicated that CHE could potentially be further developed as a novel therapeutic strategy for treatment of glioma.Copyright © 2023. Published by Elsevier Inc.