李骏飞 郑小蓉 姜建伟 章红燕

中國科学院大学附属肿瘤医院（浙江省肿瘤医院）  中国科学院基础医学与肿瘤研究所，浙江杭州   310022

[摘要] 目的 研究芍药软肝方（SRF）调控Bax/Bcl-2-Caspase-3通路抑制肝癌生长的作用及机制。 方法 建立HepG2荷瘤裸鼠模型，随机分为空白对照组、高、中、低浓度芍药软肝方（SRF）组，灌胃3周后称取瘤重，TUNEL法观察肿瘤细胞凋亡情况，Western-blot法检测对组织中NF-κB-p65、VEGF、Bcl-2、Bax、Caspase-3蛋白质表达。 结果 （1）不同浓度芍药软肝方（SRF）均可抑制HepG2荷瘤裸鼠体内肝癌肿瘤的生长，并能诱导肿瘤细胞凋亡;（2）SRF可降低裸鼠肝癌组织中NF-κB-p65、VEGF、Bcl-2蛋白表达，上调Bax、Caspase-3蛋白表达。上述作用均呈浓度依赖。 结论 SRF可显著抑制HepG2荷瘤裸鼠肿瘤生长并促进肿瘤细胞凋亡，且作用呈浓度依赖。其机制可能通过调控Bax/Bcl-2-Caspase-3通路与抑制信号通路上游NF-κB-p65及VEGF等因子的蛋白质表达有关。

[关键词] 芍药软肝方;肝癌;HepG2;分子机制

[中图分类号] R735.7          [文献标识码] A          [文章编号] 1673-9701（2021）07-0038-04

Study of the inhibitory effect of Shaoyao Ruangan Formula on the growth of liver cancer in nude mice based on Bax/Bcl-2-Caspase-3 pathway and its mechanism

LI Junfei   ZHENG Xiaorong   JIANG Jianwei   ZHANG Hongyan

The Cancer Hospital of the University of Chinese Academy of Sciences（Zhejiang Cancer Hospital）， Institute of Basic Medicine and Cancer （IBMC）， Chinese Academy of Sciences， Hangzhou  310022， China

[Abstract] Objective To study the effect and mechanism of Shaoyao Ruangan Formula（SRF） regulating Bax/Bcl-2-Caspase-3 pathway to inhibit the growth of liver cancer. Methods HepG2 tumor-bearing nude mouse models were established and randomly divided into the blank control group， the high， medium and low concentration SRF group. After three weeks of gastric perfusion， the tumors were weighed， the apoptosis of tumor cells was observed through the TUNEL method， and the protein expressions of NF-κB-p65， VEGF， Bcl-2， Bax and Caspase-3 in the tissues of different groups were detected by Western-blot method. Results （1）All SRF at variant concentrations can inhibit the growth of liver cancer in HepG2 tumor-bearing nude mice and induce the apoptosis of tumor cells; （2）SRF can decrease the protein expressions of NF-κB-p65， VEGF and Bcl-2 in liver cancer tissues of nude mice， and up-regulate the protein expressions of Bax and Caspase-3. The above-mentioned effects were all in a concentration-dependent manner. Conclusion SRF can significantly inhibit the growth of tumors in HepG2 tumor-bearing nude mice and promote the apoptosis of tumor cells in a concentration-dependent manner. The mechanism may be related to the inhibition of protein expressions of NF-κB-p65 and VEGF， etc. in the upstream of signaling pathway by regulating Bax/Bcl-2-Caspase-3 pathway.

[Key words] Shaoyao Ruangan Formula; Liver cancer; HepG2; Molecular mechanism

肝细胞癌（以下简称肝癌）是一种发病率与死亡率均较高的恶性肿瘤疾病。据统计，2018年全球肝癌患者新发约84.4万，死亡约78.2万，其新发与死亡比接近1∶1，分列36种常见癌症中的第6位及第4位[1]。我国是肝病大国，肝癌的新发率与死亡率更是常年居高不下，2015年我国肝癌患者新發39.2万余，死亡36.8万余，均位居第3位[2-4]。因此，肝癌的预防、治疗始终是当今医学界重点关注的关键问题之一。

中医中药治疗是公认的有效肝癌防治方式，亦是目前临床抗肝癌药物研发的热点之一。芍药软肝方（Shaoyao Ruangan Formula，SRF）是由我院多位中医师依据临床治疗经验共同总结、研发的中药抗肝癌协定方（当前已开发为院内中药制剂，批准文号：浙药制Z20050184。……