β,β-二甲基丙烯酰基烷宁 (DMAKN) 是中药紫草中发现的天然萘醌，是中国药典指定的定量标记物。尽管 DMAKN 在评估紫草质量方面发挥着既定的作用，但其生物潜力在研究中仍未得到充分探索。我们结合 HPLC 含量分析和综合生物信息学研究了 DMAKN 在紫草抗肝细胞癌 (HCC) 特性中的作用。随后，进行了体外和体内实验来评估DMAKN对抗HCC的功效。机制研究的重点是阐明 DMAKN 对细胞周期调节和诱导细胞死亡的影响。综合 HPLC 分析和生物信息学确定 DMAKN 是紫草抗 HCC 活性的主要活性化合物。体外和体内研究证实了 DMAKN 对抗 HCC 的强大功效。值得注意的是，DMAKN 对 HCC 细胞具有双重作用：低剂量抑制增殖，高剂量诱导细胞快速死亡。机制研究表明，低剂量 DMAKN 通过调节 CDK1 和 Cdc25C 磷酸化诱导 G2/M 期细胞周期停滞，而高剂量 DMAKN 则引发细胞坏死。重要的是，高剂量DMAKN在短时间内引起细胞内ROS水平急剧增加，而低剂量DMAKN在较长时间内逐渐增加ROS水平。此外，低剂量 DMAKN 诱导的 ROS 激活了 JNK 通路，这对细胞周期停滞至关重要，而高剂量 DMAKN 诱导的坏死是 ROS 依赖性的，但不依赖于 JNK。这项研究强调了 DMAKN 作为主要抗 HCC 化合物的关键作用在《子草》中，描述了其不同的影响和潜在机制。这些结果证明了 DMAKN 作为治疗 HCC 的治疗剂的潜力，为癌症治疗的进一步研究和进步提供了重要信息。版权所有 © 2024 作者。由 Elsevier GmbH 出版。保留所有权利。

β,β-Dimethylacrylalkannin (DMAKN), a natural naphthoquinone found in Zicao, a traditional Chinese medicine (TCM), serves as the designated quantitative marker in the Chinese Pharmacopoeia. Despite its established role in assessing Zicao quality, DMAKN's biological potential remains underexplored in research.We investigated DMAKN's involvement in Zicao's anti-hepatocellular carcinoma (HCC) properties using a combination of HPLC content analysis and comprehensive bioinformatics. Subsequently, both in vitro and in vivo experiments were conducted to evaluate DMAKN's efficacy against HCC. Mechanistic investigations focused on elucidating DMAKN's impact on cell cycle regulation and induction of cell death.Integrated HPLC analysis and bioinformatics identified DMAKN as the primary active compound responsible for Zicao's anti-HCC activity. In vitro and in vivo studies confirmed DMAKN's potent efficacy against HCC. Notably, DMAKN demonstrated dual effects on HCC cells: inhibiting proliferation at lower doses and inducing rapid cell death at higher doses. Mechanistic insights revealed that low-dose DMAKN induced G2/M phase cell cycle arrest through modulation of CDK1 and Cdc25C phosphorylation, while high-dose DMAKN triggered necrosis. Importantly, high-dose DMAKN caused a sharp increase in intracellular ROS levels in a short time, while low-dose DMAKN gradually increased ROS levels over a long period. Additionally, low-dose DMAKN-induced ROS activated the JNK pathway, crucial for cell cycle arrest, whereas high-dose DMAKN-induced necrosis was ROS-dependent but JNK-independent.This study underscores DMAKN's pivotal role as the principal anti-HCC compound in Zicao, delineating its differential effects and underlying mechanisms. These results demonstrate the potential of DMAKN as a therapeutic agent for the treatment of HCC, providing important information for further study and advancement in cancer therapy.Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.