Thioredoxin1（TRX1）是一种关键蛋白质，调节氧化还原反应，被认为是癌症治疗的关键靶点。黄酮类化合物已被证明具有较好的抗氧化和抗癌活性。本研究旨在探究黄酮类化合物缬草苷-7-葡萄糖苷（CG）通过靶向TRX1是否能够发挥抗肝癌作用。采用不同剂量的CG处理Huh-7和HepG2细胞系，计算IC50值，然后研究低、中、高剂量的CG对HCC细胞的细胞存活率、细胞凋亡、氧化应激和TRX1表达的影响。此外，采用HepG2移植瘤小鼠模型评估CG对体内HCC生长的影响。通过分子对接探讨CG和TRX1的结合模式。然后使用si-TRX1进一步探索TRX1对CG抑制HCC的效应。结果发现，CG剂量依赖性地降低了Huh-7和HepG2细胞的增殖活性，诱导细胞凋亡，显著激活了氧化应激，并抑制了TRX1的表达。体内实验也显示，CG剂量依赖性地调节了氧化应激和TRX1的表达，并促进了凋亡蛋白的表达，从而抑制了HCC生长。分子对接证实CG与TRX1有良好的结合效果。干预TRX1能够显著抑制HCC细胞的增殖，促进细胞凋亡，并进一步促进CG对细胞活性的影响。此外，CG显著增加了ROS的产生量，降低了线粒体膜电位，调节了Bax、Bcl-2和cleaved-caspase-3的表达，并激活了线粒体介导的凋亡。si-TRX1增强了CG对HCC线粒体功能和凋亡的影响，提示TRX1参与了CG抑制HCC线粒体介导凋亡的作用。综上所述，CG通过靶向TRX1调节氧化应激和促进线粒体介导的凋亡来发挥抗HCC活性。Copyright © 2023 Elsevier B.V. All rights reserved.

Thioredoxin1 (TRX1) is a key protein that regulates redox and is considered to be a key target for cancer therapy. Flavonoids have been proven to have good antioxidant and anticancer activities. This study aimed to investigate whether the flavonoid calycosin-7-glucoside (CG) exerts an anti-hepatocellular carcinoma (HCC) role by targeting TRX1. Different doses of CG were used to treat HCC cell lines Huh-7 and HepG2 to calculate the IC50. On this basis, the effects of low, medium and high doses of CG on cell viability, apoptosis, oxidative stress and TRX1 expression of HCC cells were investigated in vitro. Also, HepG2 xenograft mice were used to evaluate the role of CG on HCC growth in vivo. The binding mode of CG and TRX1 was explored by molecular docking. Then si-TRX1 was used to further discover the effects of TRX1 on CG inhibition of HCC. Results found that CG dose-dependent decreased the proliferation activity of Huh-7 and HepG2 cells, induced apoptosis, significantly activated oxidative stress and inhibited TRX1 expression. In vivo experiments also showed that CG dose-dependent regulated oxidative stress and TRX1 expression, and promoted the expression of apoptotic proteins to inhibit HCC growth. Molecular docking confirmed that CG had a good binding effect with TRX1. Intervention with TRX1 significantly inhibited the proliferation of HCC cells, promoted apoptosis, and further promoted the effect of CG on the activity of HCC cells. In addition, CG significantly increased ROS production, reduced mitochondrial membrane potential, regulated the expression of Bax, Bcl-2 and cleaved-caspase-3, and activated mitochondria-mediated apoptosis. And si-TRX1 enhanced the effects of CG on mitochondrial function and apoptosis of HCC, suggesting that TRX1 participated in the inhibitory effect of CG on mitochondria-mediated apoptosis of HCC. In conclusion, CG exerts anti-HCC activity by targeting TRX1 to regulate oxidative stress and promote mitochondria-mediated apoptosis.Copyright © 2023 Elsevier B.V. All rights reserved.