Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 28, 2024; 30(24): 3086-3105
Published online Jun 28, 2024. doi: 10.3748/wjg.v30.i24.3086
Effect and mechanism of Qingre Huashi decoction on drug-resistant Helicobacter pylori
Miao-Miao Lin, Shan-Shan Yang, Qiu-Yue Huang, Guang-Hui Cui, Xiao-Fen Jia, Yao Yang, Zong-Ming Shi, Hui Ye, Xue-Zhi Zhang
Miao-Miao Lin, Shan-Shan Yang, Qiu-Yue Huang, Guang-Hui Cui, Xiao-Fen Jia, Yao Yang, Zong-Ming Shi, Hui Ye, Xue-Zhi Zhang, Department of Integrated Traditional Chinese and Western Medicine, Peking University First Hospital, Beijing 100034, China
Miao-Miao Lin, Shan-Shan Yang, Qiu-Yue Huang, Guang-Hui Cui, Xiao-Fen Jia, Yao Yang, Zong-Ming Shi, Hui Ye, Xue-Zhi Zhang, Institute of Integrated Traditional Chinese and Western Medicine, Peking University, Beijing 100034, China
Co-corresponding authors: Hui Ye and Xue-Zhi Zhang.
Author contributions: Ye H and Zhang XZ designed the study, provided financial support, and revised the manuscript, making equal contributions to this work as co-corresponding authors; Lin MM performed the experiments, conducted data analysis, and drafted the manuscript; Jia XF, Yang Y, and Shi ZM interpreted the data; Yang SS, Huang QY, and Cui GH corrected the paper; and all authors reviewed/edited the manuscript and approved the final version.
Supported by the National Natural Science Foundation of China, No. 81973615 and No. 82304930; Natural Science Foundation of Beijing, No. 7332323; and Capital’ s Funds for Health Improvement and Research, No. CF2022-2-40711.
Institutional review board statement: The study was reviewed and approved by the Medical Ethics Committee of Peking University First Hospital (Approval No. 2022Yan130-002).
Conflict-of-interest statement: All the authors declare that they have no competing interest to disclose.
Data sharing statement: The relevant experimental data regarding humans and bacteria are available from the corresponding authors on reasonable request.
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: Xue-Zhi Zhang, MD, PhD, Chief Physician, Professor, Department of Integrated Traditional Chinese and Western Medicine, Peking University First Hospital, No. 8 Xishiku Street, Xicheng District, Beijing 100034, China. zhang.xuezhi@263.net
Received: February 25, 2024
Revised: May 5, 2024
Accepted: May 29, 2024
Published online: June 28, 2024
Processing time: 121 Days and 4.8 Hours
Abstract
BACKGROUND

Helicobacter pylori (HP), the most common pathogenic microorganism in the stomach, can induce inflammatory reactions in the gastric mucosa, causing chronic gastritis and even gastric cancer. HP infection affects over 4.4 billion people globally, with a worldwide infection rate of up to 50%. The multidrug resistance of HP poses a serious challenge to eradication. It has been de-monstrated that compared to bismuth quadruple therapy, Qingre Huashi decoction (QHD) combined with triple therapy exhibits comparable eradication rates but with a lower incidence of adverse reactions; in addition, QHD can directly inhibit and kill HP in vitro.

AIM

To explore the effect and mechanism of QHD on clinically multidrug-resistant and strong biofilm-forming HP.

METHODS

In this study, 12 HP strains were isolated in vitro after biopsy during gastroscopy of HP-infected patients. In vitro, the minimum inhibitory concentration (MIC) values for clinical HP strains and biofilm quantification were determined through the E-test method and crystal violet staining, respectively. The most robust biofilm-forming strain of HP was selected, and QHD was evaluated for its inhibitory and bactericidal effects on the strain with strong biofilm formation. This assessment was performed using agar dilution, E-test, killing dynamics, and transmission electron microscopy (TEM). The study also explored the impact of QHD on antibiotic resistance in these HP strains with strong biofilm formation. Crystalline violet method, scanning electron microscopy, laser confocal scanning microscopy, and (p)ppGpp chromatographic identification were employed to evaluate the effect of QHD on biofilm in strong biofilm-forming HP strains. The effect of QHD on biofilm and efflux pump-related gene expression was evaluated by quantitative polymerase chain reaction. Non-targeted metabolomics with UHPLC-MS/MS was used to identify potential metabolic pathways and biomarkers which were different between the NC and QHD groups.

RESULTS

HP could form biofilms of different degrees in vitro, and the intensity of formation was associated with the drug resistance of the strain. QHD had strong bacteriostatic and bactericidal effects on HP, with MICs of 32-64 mg/mL. QHD could inhibit the biofilm formation of the strong biofilm-forming HP strains, disrupt the biofilm structure, lower the accumulation of (p)ppGpp, decrease the expression of biofilm-related genes including LuxS, Spot, glup (HP1174), NapA, and CagE, and reduce the expression of efflux pump-related genes such as HP0605, HP0971, HP1327, and HP1489. Based on metabolomic analysis, QHD induced oxidative stress in HP, enhanced metabolism, and potentially inhibited relevant signaling pathways by upregulating adenosine monophosphate (AMP), thereby affecting HP growth, metabolism, and protein synthesis.

CONCLUSION

QHD exerts bacteriostatic and bactericidal effects on HP, and reduces HP drug resistance by inhibiting HP biofilm formation, destroying its biofilm structure, inhibiting the expression of biofilm-related genes and efflux pump-related genes, enhancing HP metabolism, and activating AMP in HP.

Keywords: Qingre Huashi decoction, Helicobacter pylori, Drug resistance, Biofilm, Metabolomics

Core Tip:Helicobacter pylori (HP) infection affects over 4.4 billion people worldwide, with its multidrug resistance posing a serious challenge. Twelve strains of HP were isolated in vitro following biopsy during gastroscopy of HP-infected patients, with the most robust biofilm-forming strain of HP being selected to investigate the effect and mechanism of Qingre Huashi decoction (QHD) on drug-resistant HP. QHD has bacteriostatic and bactericidal effects on HP, reduces HP drug-resistance by inhibiting biofilm formation, destroying biofilm structure, inhibiting biofilm-related and efflux pump-related gene expression, enhancing HP metabolism, and activating adenosine monophosphate in HP.