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Volume 42 Issue 1

Jan. 2026

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Article Navigation > Journal of Clinical Hepatology > 2026 > 42(1): 125-133

Effect and mechanism of Biejiajian Pill on subcutaneous xenograft tumor model of hepatocellular carcinoma Huh7 cells

DOI: 10.12449/JCH260115

1.
Graduate School of Guangxi University of Chinese Medicine，Nanning 530022，China
2.
Department of Spleen and Stomach Hepatology，Xianhu Branch of The First Affiliated Hospital of Guangxi University of Chinese Medicine，Nanning 530022，China
3.
Guangxi Key Laboratory of Translational Medicine for Treating High-Incidence Infectious Diseases with Integrative Medicine，Nanning 530022，China
4.
Department of Chinese Internal Medicine，The First Affiliated Hospital of Guangxi University of Chinese Medicine，Nanning 530022，China
5.
Department of Hepatology，The First Affiliated Hospital of Guangxi University of Chinese Medicine，Nanning 530022，China

Research funding:

National Natural Science Foundation of China (8246150205);

Guangxi Natural Science Foundation (2022GXNSFAA035460);

Guangxi Natural Science Foundation (2024GXNSFDA010005)

More Information

Corresponding author: HUANG Jingjing， 55869563@qq.com （ORCID： 0000-0002-4932-0838）
Received Date: 2025-07-12
Accepted Date: 2025-10-15
Published Date: 2026-01-25

Abstract

Objective To investigate the inhibitory effect of Biejiajian Pills （BJJW） on the growth of liver cancer， as well as its potential mechanism in mediating the AMP-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） pathway through mitochondrial energy metabolism. Methods Human hepatoma Huh7 cells were used to establish a nude mouse model of subcutaneous xenograft tumor. A total of 18 tumor-bearing nude mice were randomly divided into model group， BJJW group （2.2 g/kg）， and metformin group （250 mg/kg）， and the corresponding drug was given by gavage for 14 consecutive days. Tumor volume and weight were monitored during the experiment； HE staining was used to observe histopathological changes； the levels of reactive oxygen species （ROS） and adenosine triphosphate （ATP） in tumor tissue were measured； immunohistochemistry and Western blotting were used to measure the expression levels of proteins associated with the AMPK/mTOR pathway. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups， and the Tukey’s test was used for further comparison between two groups； the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups， and the Dunn’s test was used for further comparison between two groups. Results Compared with the model group， the BJJW group had a tumor inhibition rate of 45.73%， with significant reductions in both tumor volume and weight （P<0.01）. Pathological examination showed that compared with the model group， the BJJW group had a significant reduction in the number of tumor cells and the presence of extensive necrosis. Mechanistic studies showed that compared with the model group， the BJJW group had a significant increase in ROS level （P<0.001） and a significant reduction in ATP level （P<0.001）， as well as significant increases in p-AMPK/AMPK ratio （0.81±0.20 vs 0.13±0.04， P<0.01） and p-ULK1/ULK1 ratio （0.69±0.17 vs 0.18±0.13， P<0.01） and a significant reduction in p-mTOR/mTOR ratio （1.34±0.16 vs 3.20±0.62， P<0.01）. Conclusion BJJW may inhibit the growth of liver cancer by inducing mitochondrial energy metabolism dysfunction， increasing the level of ROS， reducing the level of ATP， and activating the AMPK/mTOR signaling pathway.
- Biejiajian Pills,
- Liver Neoplasms,
- AMP-activated Protein Kinase,
- Mammalian Target of Rapamycin

References

[1]	DENG XX, LI H, ZHONG YR, et al. Burden of liver cancer attributable to hepatitis B and alcohol globally, in China, and for five sociodemographic index regions from 1990 to 2021: A population-based study[J]. J Clin Transl Hepatol, 2025, 13( 1): 1- 14. DOI: 10.14218/JCTH.2024.00351.
[2]	ZHOU Y, DING JM, QIN ZY, et al. Predicting the survival rate of patients with hepatocellular carcinoma after thermal ablation by nomograms[J]. Ann Transl Med, 2020, 8( 18): 1159. DOI: 10.21037/atm-20-6116.
[3]	CAO MD, XIA CF, CAO MM, et al. Attributable liver cancer deaths and disability-adjusted life years in China and worldwide: Profiles and changing trends[J]. Cancer Biol Med, 2024, 21( 8): 679- 691. DOI: 10.20892/j.issn.2095-3941.2024.0149.
[4]	LIU Y, ZHENG JX, HAO JL, et al. Global burden of primary liver cancer by five etiologies and global prediction by 2035 based on global burden of disease study 2019[J]. Cancer Med, 2022, 11( 5): 1310- 1323. DOI: 10.1002/cam4.4551.
[5]	YU Z, MAN XW, JIANG Y, et al. Analysis of hospitalization expenses and influencing factors of 5 487 patients with liver cancer in Beijing[J]. Chin J Health Stat, 2020, 37( 5): 748- 751. DOI: 10.3969/j.issn.1002-3674.2020.05.029. 于哲, 满晓玮, 蒋艳, 等. 北京市5 487例肝癌患者住院费用及影响因素分析[J]. 中国卫生统计, 2020, 37( 5): 748- 751. DOI: 10.3969/j.issn.1002-3674.2020.05.029.
[6]	RANKOVIĆ B, HAUPTMAN N. Circulating microRNA panels for detection of liver cancers and liver-metastasizing primary cancers[J]. Int J Mol Sci, 2023, 24( 20): 15451. DOI: 10.3390/ijms242015451.
[7]	ZENG LL, ZHU LT, FU SS, et al. Mitochondrial dysfunction-molecular mechanisms and potential treatment approaches of hepatocellular carcinoma[J]. Mol Cell Biochem, 2025, 480( 4): 2131- 2142. DOI: 10.1007/s11010-024-05144-4.
[8]	FAUBERT B, SOLMONSON A, DEBERARDINIS RJ. Metabolic reprogramming and cancer progression[J]. Science, 2020, 368( 6487): eaaw5473. DOI: 10.1126/science.aaw5473.
[9]	LI YF, XU JJ, LAN T, et al. Biejiajian Pill inhibits the malignant biological behavior of Hep3B cells through CMTM6[J]. J Xi’an Jiaotong Univ Med Sci, 2025, 46( 3): 522- 531. DOI: 10.7652/jdyxb202503021. 李尹凡, 徐隽婕, 兰涛, 等. 鳖甲煎丸通过CMTM6抑制Hep3B细胞的恶性生物学行为[J]. 西安交通大学学报(医学版), 2025, 46( 3): 522- 531. DOI: 10.7652/jdyxb202503021.
[10]	SHAO FL, CHEN QP, BI Q, et al. Intervention mechanism of Biejiajian Wan on primary liver cancer by regulating lncRNA SNHG5/miRNA-26a-5p/GSK-3β signal axis[J]. Chin J Exp Tradit Med Formulae, 2024, 30( 4): 107- 113. DOI: 10.13422/j.cnki.syfjx.20230730. 邵范雷, 陈秋平, 毕倩, 等. 鳖甲煎丸调控lncRNA SNHG5/miRNA-26a-5p/GSK-3β信号轴干预原发性肝癌的作用机制[J]. 中国实验方剂学杂志, 2024, 30( 4): 107- 113. DOI: 10.13422/j.cnki.syfjx.20230730.
[11]	FENG MH, HE SQ, HUANG SZ, et al. Inhibitory effect of Biejiajian pills against diethylnitrosamine-induced hepatocarcinogenesis in rats[J]. J South Med Univ, 2020, 40( 8): 1148- 1154. DOI: 10.12122/j.issn.1673-4254.2020.08.12. 冯明辉, 贺松其, 黄松泽, 等. 鳖甲煎丸对二乙基亚硝胺诱导的大鼠肝癌的抑制作用及机制[J]. 南方医科大学学报, 2020, 40( 8): 1148- 1154. DOI: 10.12122/j.issn.1673-4254.2020.08.12.
[12]	YU XP, WANG SY, MENG QG, et al. Exploring regulatory mechanism of Biejiajian pill on hepatocellular carcinoma based on bioinformatics[J]. Chin Arch Tradit Chin Med, 2025, 43( 4): 82- 89. DOI: 10.13193/j.issn.1673-7717.2025.04.015. 于欣萍, 王诗尧, 孟庆刚, 等. 基于生物信息学探究鳖甲煎丸对肝细胞癌的调控机制[J]. 中华中医药学刊, 2025, 43( 4): 82- 89. DOI: 10.13193/j.issn.1673-7717.2025.04.015.
[13]	National Institutes of Health Office of Laboratory Animal Welfare. Animal use and management[EB/OL].( 2024-11-25)[ 2025-04-22]. https://olaw.nih.gov/guidance/topic-index/animal-use.htm. https://olaw.nih.gov/guidance/topic-index/animal-use.htm
[14]	KUAI Z, CHAO X, HE YT, et al. Metformin attenuates inflammation and boosts autophagy in the liver and intestine of chronologically aged rats[J]. Exp Gerontol, 2023, 184: 112331. DOI: 10.1016/j.exger.2023.112331.
[15]	WANG GF, GAO H, DAI SN, et al. Metformin inhibits neutrophil extracellular traps-promoted pancreatic carcinogenesis in obese mice[J]. Cancer Lett, 2023, 562: 216155. DOI: 10.1016/j.canlet.2023.216155.
[16]	YENMIŞ G, BEŞLI N, YAPRAK SARAÇ E, et al. Metformin promotes apoptosis in primary breast cancer cells by downregulation of cyclin D1 and upregulation of P53 through an AMPK-alpha independent mechanism[J]. Turk J Med Sci, 2021, 51( 2): 826- 834. DOI: 10.3906/sag-1908-112.
[17]	DAS BK, KNOTT RM, GADAD PC. Metformin and asarone inhibit HepG2 cell proliferation in a high glucose environment by regulating AMPK and Akt signaling pathway[J]. Future J Pharm Sci, 2021, 7( 1): 43. DOI: 10.1186/s43094-021-00193-8.
[18]	LUO S, SCHOOLING CM, WONG ICK, et al. Evaluating the impact of AMPK activation, a target of metformin, on risk of cardiovascular diseases and cancer in the UK Biobank: A Mendelian randomisation study[J]. Diabetologia, 2020, 63( 11): 2349- 2358. DOI: 10.1007/s00125-020-05243-z.
[19]	TAN QW, XU J, ZHU RH, et al. Effects of Biejia Jianwan on the tumorigenicity and dryness of liver cancer stem cells by regulating miR-140[J]. Jilin J Chin Med, 2024, 44( 1): 90- 95. DOI: 10.13463/j.cnki.jlzyy.2024.01.021. 谭钦文, 徐健, 朱荣火, 等. 鳖甲煎丸调控miR-140对肝癌干细胞致瘤性及干性的影响[J]. 吉林中医药, 2024, 44( 1): 90- 95. DOI: 10.13463/j.cnki.jlzyy.2024.01.021.
[20]	TAN QW, HUANG JJ, ZHONG RX, et al. Effect of Biejia Decoction Pill on aerobic glycolysis in hepatocellular carcinoma by regulating the protein kinase B/mammalian target of rapamycin signaling pathway[J]. J Clin Hepatol, 2025, 41( 2): 300- 306. DOI: 10.12449/JCH250216. 谭钦文, 黄晶晶, 钟瑞熙, 等. 鳖甲煎丸调控AKT/mTOR信号通路在肝癌细胞有氧糖酵解中的作用[J]. 临床肝胆病杂志, 2025, 41( 2): 300- 306. DOI: 10.12449/JCH250216.
[21]	LIN HS, TAN JN, FANG QL, et al. Analysis of the circRNA expression profile in hepatocellular carcinoma cells inhibited by Biejia Jianwan[J]. Genom Appl Biol, 2025, 44( 5): 511- 519. DOI: 10.13417/j.gab.044.000511. 林洪升, 谭金娜, 方巧玲, 等. 鳖甲煎丸抑制肝癌细胞的环状RNA表达谱分析[J]. 基因组学与应用生物学, 2025, 44( 5): 511- 519. DOI: 10.13417/j.gab.044.000511.
[22]	CHEN WG, HE CY, WEN B, et al. Biejiajian pill regulates ferroptosis in hepatocellular carcinoma cells via p62/Keap1/NRF2 signaling pathway: A mechanism study[J]. J Sichuan Univ Med Sci, 2025, 56( 1): 51- 58. DOI: 10.12182/20250160502. 陈伟光, 何春雨, 文彬, 等. 鳖甲煎丸通过p62/Keap1/NRF2信号通路调控肝癌细胞铁死亡的作用机制研究[J]. 四川大学学报(医学版), 2025, 56( 1): 51- 58. DOI: 10.12182/20250160502.
[23]	SUN JL, WEN B, YANG XM, et al. Mechanism research of regulation of proliferation and metastasis of hepatoma cells Hep3B by Biejiajian Pill based on PI3K/AKT/GSK-3β signaling pathway[J]. China J Tradit Chin Med Pharm, 2021, 36( 3): 1361- 1365. 孙嘉玲, 文彬, 杨雪梅, 等. 基于PI3K/AKT/GSK-3β信号通路探讨鳖甲煎丸调控肝癌细胞Hep3B增殖转移的机制[J]. 中华中医药杂志, 2021, 36( 3): 1361- 1365.
[24]	HUANG JJ, HUANG HN, WANG XJ, et al. Bie Jia Jian pill enhances the amelioration of bone mesenchymal stem cells on hepatocellular carcinoma progression[J]. J Nat Med, 2022, 76( 1): 49- 58. DOI: 10.1007/s11418-021-01548-4.
[25]	WANG WL, SUN Q, WU ZH, et al. Mitochondrial dysfunction-related genes in hepatocellular carcinoma[J]. Front Biosci, 2013, 18( 3): 1141- 1149. DOI: 10.2741/4169.
[26]	LEE HY, NGA HT, TIAN JW, et al. Mitochondrial metabolic signatures in hepatocellular carcinoma[J]. Cells, 2021, 10( 8): 1901. DOI: 10.3390/cells10081901.
[27]	van der BLIEK AM, SEDENSKY MM, MORGAN PG. Cell biology of the mitochondrion[J]. Genetics, 2017, 207( 3): 843- 871. DOI: 10.1534/genetics.117.300262.
[28]	HU CX, HUANG Y, LI LJ. Drp1-dependent mitochondrial fission plays critical roles in physiological and pathological progresses in mammals[J]. Int J Mol Sci, 2017, 18( 1): 144. DOI: 10.3390/ijms18010144.
[29]	SHADEL GS, HORVATH TL. Mitochondrial ROS signaling in organismal homeostasis[J]. Cell, 2015, 163( 3): 560- 569. DOI: 10.1016/j.cell.2015.10.001.
[30]	TIAN CW, LIU YF, LI ZS, et al. Mitochondria related cell death modalities and disease[J]. Front Cell Dev Biol, 2022, 10: 832356. DOI: 10.3389/fcell.2022.832356.
[31]	HSU CC, PENG DN, CAI Z, et al. AMPK signaling and its targeting in cancer progression and treatment[J]. Semin Cancer Biol, 2022, 85: 52- 68. DOI: 10.1016/j.semcancer.2021.04.006.
[32]	TREFTS E, SHAW RJ. AMPK: Restoring metabolic homeostasis over space and time[J]. Mol Cell, 2021, 81( 18): 3677- 3690. DOI: 10.1016/j.molcel.2021.08.015.
[33]	FANG GX, ZHANG PL, LIU JF, et al. Inhibition of GSK-3β activity suppresses HCC malignant phenotype by inhibiting glycolysis via activating AMPK/mTOR signaling[J]. Cancer Lett, 2019, 463: 11- 26. DOI: 10.1016/j.canlet.2019.08.003.
[34]	LIU YF, XU YY, WANG F, et al. Inhibition of AMPK activity by TRIM11 facilitates cell survival of hepatocellular carcinoma under metabolic stress[J]. Clin Transl Med, 2021, 11( 12): e617. DOI: 10.1002/ctm2.617.
[35]	MENG SS, GU HW, ZHANG T, et al. Gradual deterioration of fatty liver disease to liver cancer via inhibition of AMPK signaling pathways involved in energy-dependent disorders, cellular aging, and chronic inflammation[J]. Front Oncol, 2023, 13: 1099624. DOI: 10.3389/fonc.2023.1099624.
[36]	JUNG TY, RYU JE, JANG MM, et al. Naa20, the catalytic subunit of NatB complex, contributes to hepatocellular carcinoma by regulating the LKB1-AMPK-mTOR axis[J]. Exp Mol Med, 2020, 52( 11): 1831- 1844. DOI: 10.1038/s12276-020-00525-3.
[37]	ZHOU XJ, CHEN Y, WANG FF, et al. Artesunate induces autophagy dependent apoptosis through upregulating ROS and activating AMPK-mTOR-ULK1 axis in human bladder cancer cells[J]. Chem Biol Interact, 2020, 331: 109273. DOI: 10.1016/j.cbi.2020.109273.
[38]	JIA L, LIN XR, GUO WY, et al. Salvia chinensia Benth induces autophagy in esophageal cancer cells via AMPK/ULK1 signaling pathway[J]. Front Pharmacol, 2022, 13: 995344. DOI: 10.3389/fphar.2022.995344.
[39]	NAIK PP, MUKHOPADHYAY S, PRAHARAJ PP, et al. Secretory clusterin promotes oral cancer cell survival via inhibiting apoptosis by activation of autophagy in AMPK/mTOR/ULK1 dependent pathway[J]. Life Sci, 2021, 264: 118722. DOI: 10.1016/j.lfs.2020.118722.
[40]	WANG CM, CIGLIANO A, JIANG LJ, et al. 4EBP1/eIF4E and p70S6K/RPS6 axes play critical and distinct roles in hepatocarcinogenesis driven by AKT and N-Ras proto-oncogenes in mice[J]. Hepatology, 2015, 61( 1): 200- 213. DOI: 10.1002/hep.27396.
[41]	XU M, WANG ZJ, WU ZS, et al. Autophagy activated by the AMPK/mTOR/ULK1 pathway involves AURKB-mediated microgliosis in neuropathic pain[J]. Brain Behav Immun, 2025, 129: 948- 959. DOI: 10.1016/j.bbi.2025.07.019.

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