Department of Pediatrics, The Affiliated Hospital of Southwest Medical University, Sichuan Clinical Research Center for Birth Defects, Luzhou, Sichuan, China.
State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China.
PLoS One. 2023 Oct 10;18(10):e0289697. doi: 10.1371/journal.pone.0289697. eCollection 2023.
The unmet needs in treating acute myeloid leukemia(AML) promote us to look for more effective and less toxic therapies. In this study, we discovered that Yinzhihuang injection(YZHI), a traditional Chinese patent medicine for hepatitis treatment, suppressed the growth of AML cells.
Anti-proliferative activities of YZHI were measured by CCK-8 assay. Cell cycle arrest was evaluated by PI staining, and apoptosis was evaluated by annexin V/PI staining. To explore the cell cycle arrest and cell death mechanism induced by YZHI, we assessed a series of assays, including measurements of the protein expression and cellular ATP. The anti-tumor activity was further demonstrated in nude mice.
Flow cytometric and biochemical analysis revealed that YZHI caused cell cycle arrest and induced apoptosis in the AML HL-60 cells. Mechanistically, YZHI activated AMPK by promoting phosphorylation of the kinase. The active AMPK negatively regulated the downstream target mTORC1, leading to the inhibition of cell proliferation and induction of apoptosis. Pretreatment with the AMPK inhibitor compound C rescued YZHI induced apoptosis and partially restored cell proliferation of HL-60. Consistent with the data in vitro, YZHI obviously suppressed subcutaneous xenograft growth in nude mice.
In a word, our data suggest that YZHI can be repurposed for the treatment of AML, which is worthy of further clinical evaluation.
治疗急性髓系白血病（AML）的未满足需求促使我们寻找更有效和更少毒性的治疗方法。在这项研究中，我们发现茵栀黄注射液（YZHI），一种治疗肝炎的中药专利药物，能抑制 AML 细胞的生长。
通过 CCK-8 法测定 YZHI 的抗增殖活性。通过 PI 染色评估细胞周期停滞，通过 Annexin V/PI 染色评估细胞凋亡。为了探索 YZHI 诱导的细胞周期停滞和细胞死亡机制，我们评估了一系列实验，包括蛋白表达和细胞内 ATP 的测量。在裸鼠中进一步证明了抗肿瘤活性。
流式细胞术和生化分析显示，YZHI 导致 AML HL-60 细胞周期停滞并诱导细胞凋亡。在机制上，YZHI 通过促进激酶磷酸化激活 AMPK。活性 AMPK 负调控下游靶标 mTORC1，从而抑制细胞增殖并诱导细胞凋亡。用 AMPK 抑制剂化合物 C 预处理可挽救 YZHI 诱导的凋亡，并部分恢复 HL-60 的细胞增殖。与体外数据一致，YZHI 明显抑制裸鼠皮下异种移植瘤的生长。
总之，我们的数据表明 YZHI 可重新用于治疗 AML，值得进一步临床评估。
