Hernandezine (Her) 是一种从唐松草中提取的双苄基异喹啉生物碱，因其草药固有的一系列生物活性而受到认可。尽管其具有显着的特性，但其抗癌作用在很大程度上仍未被探索。在这项研究中，我们阐明了 Her 通过激活细胞凋亡和坏死性凋亡机制显着诱导癌细胞的细胞毒性。此外，Her 通过激活 AMPK 和 ATG5 接合系统触发自噬体形成，导致 LC3 脂化。我们的研究结果表明，Her 对线粒体膜造成损伤，受损的线粒体发生线粒体自噬，线粒体自噬标记物表达升高证明了这一点。相反，Her 破坏了自噬流，这通过 p62 的上调和自溶酶体的积累来证明，如 RFP-GFP-LC3 报告基因测定中所观察到的。最初，我们确定 Her 并没有阻止自噬体和溶酶体的融合。然而，它抑制组织蛋白酶 D 的成熟并增加溶酶体 pH 值，表明溶酶体功能受损。使用早期自噬抑制剂 3-甲基腺嘌呤 (3-MA) 不会抑制 LC3II，这表明 Her 还在自噬体形成中诱导非典型自噬。 Bafilomycin A1（一种非经典自噬抑制剂）的应用减少了 Her 对 ATG16L1 的募集和 LC3II 的积累，从而增加了 Her 诱导的细胞死亡。这些观察结果表明，虽然自噬最初发挥保护作用，但 Her 对自噬过程的破坏会促进程序性细胞死亡。这项研究提供了第一个证据，证明 Her 在诱导细胞凋亡和坏死性凋亡中发挥双重作用，同时还启动并随后损害自噬以促进细胞凋亡。这些见解有助于更深入地了解程序性细胞死亡的机制，为增强癌症预防和治疗策略提供潜在途径。版权所有 © 2024 中国药科大学。由 Elsevier B.V. 出版。保留所有权利。

Hernandezine (Her), a bisbenzylisoquinoline alkaloid extracted from Thalictrum flavum, is recognized for its range of biological activities inherent to this herbal medicine. Despite its notable properties, the anti-cancer effects of Her have remained largely unexplored. In this study, we elucidated that Her significantly induced cytotoxicity in cancer cells through the activation of apoptosis and necroptosis mechanisms. Furthermore, Her triggered autophagosome formation by activating the AMPK and ATG5 conjugation systems, leading to LC3 lipidation. Our findings revealed that Her caused damage to the mitochondrial membrane, with the damaged mitochondria undergoing mitophagy, as evidenced by the elevated expression of mitophagy markers. Conversely, Her disrupted autophagic flux, demonstrated by the upregulation of p62 and accumulation of autolysosomes, as observed in the RFP-GFP-LC3 reporter assay. Initially, we determined that Her did not prevent the fusion of autophagosomes and lysosomes. However, it inhibited the maturation of cathepsin D and increased lysosomal pH, indicating an impairment of lysosomal function. The use of the early-stage autophagy inhibitor, 3-methyladenine (3-MA), did not suppress LC3II, suggesting that Her also induces noncanonical autophagy in autophagosome formation. The application of Bafilomycin A1, an inhibitor of noncanonical autophagy, diminished the recruitment of ATG16L1 and the accumulation of LC3II by Her, thereby augmenting Her-induced cell death. These observations imply that while autophagy initially plays a protective role, the disruption of the autophagic process by Her promotes programmed cell death. This study provides the first evidence of Her's dual role in inducing apoptosis and necroptosis while also initiating and subsequently impairing autophagy to promote apoptotic cell death. These insights contribute to a deeper understanding of the mechanisms underlying programmed cell death, offering potential avenues for enhancing cancer prevention and therapeutic strategies.Copyright © 2024 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.