Chongqing Academy of Chinese Materia Medica, Chongqing 400065, PR China; Sichuan-Chongqing Joint Key Laboratory of Innovation of New Drugs of Traditional Chinese Medicine, Chongqing 400065, PR China.
State Key Laboratory of Chinese Medicine and Molecular Pharmacology (Incubation), The Hong Kong Polytechnic University Shenzhen Research Institute, Shenzhen 518057, PR China; Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, PR China.
Phytomedicine. 2024 Nov;134:155959. doi: 10.1016/j.phymed.2024.155959. Epub 2024 Aug 19.
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.
β,β-二甲基丙烯酰阿卡宁（DMAKN）是一种天然萘醌，存在于中药紫草中，是《中国药典》中指定的定量标志物。尽管它在评估紫草质量方面发挥了作用，但 DMAKN 的生物学潜力在研究中仍未得到充分探索。
我们采用 HPLC 含量分析和综合生物信息学相结合的方法，研究了 DMAKN 在紫草抗肝癌（HCC）特性中的作用。随后，我们进行了体内外实验来评估 DMAKN 对 HCC 的疗效。机制研究集中在阐明 DMAKN 对细胞周期调控和诱导细胞死亡的影响。
综合 HPLC 分析和生物信息学鉴定出 DMAKN 是紫草抗 HCC 活性的主要活性化合物。体内外研究证实了 DMAKN 对 HCC 的强大疗效。值得注意的是，DMAKN 对 HCC 细胞表现出双重作用：低剂量时抑制增殖，高剂量时快速诱导细胞死亡。机制研究揭示，低剂量 DMAKN 通过调节 CDK1 和 Cdc25C 磷酸化诱导 G2/M 期细胞周期阻滞，而高剂量 DMAKN 则触发细胞坏死。重要的是，高剂量 DMAKN 在短时间内急剧增加细胞内 ROS 水平，而低剂量 DMAKN 则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，这对细胞周期阻滞至关重要，而高剂量 DMAKN 诱导的坏死则依赖于 ROS，但不依赖于 JNK。
本研究强调了 DMAKN 作为紫草中主要抗 HCC 化合物的关键作用，阐述了其不同的作用和潜在机制。这些结果表明 DMAKN 作为 HCC 治疗的治疗剂具有潜力，为癌症治疗的进一步研究和进展提供了重要信息。
