Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 7, 2024; 30(13): 1911-1925
Published online Apr 7, 2024. doi: 10.3748/wjg.v30.i13.1911
Inhibition of hepatitis B virus via selective apoptosis modulation by Chinese patent medicine Liuweiwuling Tablet
Fei-Lin Ge, Yan Yang, Lan-Lan Si, Yuan-Hua Li, Meng-Zhen Cao, Jun Wang, Zhao-Fang Bai, Zhi-Gang Ren, Xiao-He Xiao, Yan Liu
Fei-Lin Ge, Department of Chinese Medicine, State Key Laboratory of Antiviral Drugs, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
Fei-Lin Ge, Lan-Lan Si, Yuan-Hua Li, Meng-Zhen Cao, Zhao-Fang Bai, Xiao-He Xiao, Yan Liu, The Fifth Medical Center of Chinese PLA General Hospital, Beijing 100039, China
Yan Yang, College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing 400010, China
Jun Wang, Beijing Key Laboratory of Emerging Infectious Diseases, Peking University Ditan Teaching Hospital, Beijing 100015, China
Zhi-Gang Ren, Department of Infectious Diseases, State Key Laboratory of Antiviral Drugs, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
Co-first authors: Fei-Lin Ge and Yan Yang.
Co-corresponding authors: Yan Liu and Xiao-He Xiao.
Author contributions: Liu Y and Xiao XH made substantial contributions to the conception or design of the study; Ge FL and Yang Y were in charge of the acquisition, analysis and interpretation of data, and wrote the manuscript; Li YH, Cao MZ and Wang J participated in experiments related to cell and animal models; Bai ZF, Ren ZG and Si LL were responsible for revision of the manuscript. All authors finally read and approved the version to be published. Ge FL and Yang Y contributed equally to this work as co-first authors. The reasons are the following. First, the research was performed as a collaborative effort, and the designation of co-first authors authorship accurately reflects the distribution of responsibilities and burdens associated with the time and effort required to complete the study and the resultant paper. Second, co-first authors contributed efforts of equal substance throughout the research process. Liu Y and Xiao XH contributed equally to this work as co-corresponding authors. The reasons are the following. First, they played a key role in coordinating the research team. Second, they made a great contribution to the original innovation of the article. In summary, we believe that designating Ge FL and Yang Y as co-first authors, Liu Y and Xiao XH as co-corresponding authors is fitting for our manuscript as it accurately reflects our team’s collaborative spirit, equal contributions, and diversity.
Supported by National Natural Science Foundation of China, No. 81930110; The National Funded Postdoctoral Researcher Program of China, No. GZC20232406; Henan Province Traditional Chinese Medicine Science Research Project, No. 2023ZY3040; Henan Province Medical Science and Technology Research Plan Joint Construction Project, No. LHGJ20230233; and National Key Research and Development Program of China, No. 2022YFC2303103.
Institutional animal care and use committee statement: Our research had been approved by the Committee on the Ethics of Animal Experiments of the Fifth Medical Center of Chinese PLA General Hospital (Permit number: IACUC-2021-0009).
Conflict-of-interest statement: The authors of this manuscript having no conflicts of interest to disclose.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Yan Liu, PhD, The Fifth Medical Center of Chinese PLA General Hospital, No. 100 Xisihuan Middle Road, Beijing 100039, China. liuyan5360@163.com
Received: October 10, 2023
Peer-review started: October 10, 2023
First decision: December 27, 2023
Revised: January 7, 2024
Accepted: February 25, 2024
Article in press: February 25, 2024
Published online: April 7, 2024
Processing time: 176 Days and 4.8 Hours
Abstract
BACKGROUND

Liuweiwuling Tablet (LWWL) is a Chinese patent medicine approved for the treatment of chronic inflammation caused by hepatitis B virus (HBV) infection. Previous studies have indicated an anti-HBV effect of LWWL, specifically in terms of antigen inhibition, but the underlying mechanism remains unclear.

AIM

To investigate the potential mechanism of action of LWWL against HBV.

METHODS

In vitro experiments utilized three HBV-replicating and three non-HBV-replicating cell lines. The in vivo experiment involved a hydrodynamic injection-mediated mouse model with HBV replication. Transcriptomics and metabolomics were used to investigate the underlying mechanisms of action of LWWL.

RESULTS

In HepG2.1403F cells, LWWL (0.8 mg/mL) exhibited inhibitory effects on HBV DNA, hepatitis B surface antigen and pregenomic RNA (pgRNA) at rates of 51.36%, 24.74% and 50.74%, respectively. The inhibition rates of LWWL (0.8 mg/mL) on pgRNA/covalently closed circular DNA in HepG2.1403F, HepG2.2.15 and HepG2.A64 cells were 47.78%, 39.51% and 46.74%, respectively. Integration of transcriptomics and metabolomics showed that the anti-HBV effect of LWWL was primarily linked to pathways related to apoptosis (PI3K-AKT, CASP8-CASP3 and P53 pathways). Apoptosis flow analysis revealed that the apoptosis rate in the LWWL-treated group was significantly higher than in the control group (CG) among HBV-replicating cell lines, including HepG2.2.15 (2.92% ± 1.01% vs 6.68% ± 2.04%, P < 0.05), HepG2.A64 (4.89% ± 1.28% vs 8.52% ± 0.50%, P < 0.05) and HepG2.1403F (3.76% ± 1.40% vs 7.57% ± 1.35%, P < 0.05) (CG vs LWWL-treated group). However, there were no significant differences in apoptosis rates between the non-HBV-replicating HepG2 cells (5.04% ± 0.74% vs 5.51% ± 1.57%, P > 0.05), L02 cells (5.49% ± 0.80% vs 5.48% ± 1.01%, P > 0.05) and LX2 cells (6.29% ± 1.54% vs 6.29% ± 0.88%, P > 0.05). TUNEL staining revealed a significantly higher apoptosis rate in the LWWL-treated group than in the CG in the HBV-replicating mouse model, while no noticeable difference in apoptosis rates between the two groups was observed in the non-HBV-replicating mouse model.

CONCLUSION

Preliminary results suggest that LWWL exerts a potent inhibitory effect on wild-type and drug-resistant HBV, potentially involving selective regulation of apoptosis. These findings offer novel insights into the anti-HBV activities of LWWL and present a novel mechanism for the development of anti-HBV medications.

Keywords: Hepatitis B virus; Chinese patent medicine; Antiviral activity; Antiviral mechanism; Selective apoptosis

Core Tip: Liuweiwuling Tablet (LWWL) exerts a potent inhibitory effect on both wild-type and drug-resistant hepatitis B virus (HBV) models. This effect appears to hinge on selective apoptosis regulation-an unprecedented discovery in the realm of anti-HBV drug mechanisms. These revelatory findings not only deepen our understanding of LWWL’s anti-HBV properties but also offer an innovative mechanism to inspire the development of new anti-HBV drugs.