In vitro anti- Helicobacter pylori activity and the underlining mechanism of an empirical herbal formula - Hezi Qingyou

Zhong Feng; Hui Li; Yajie Hao; Chang Peng; Ling Ou; Junwei Jia; Mingjin Xun; Yuanjing Zou; Meiyun Chen; Guimin Zhang; Meicun Yao

doi:10.3389/fmicb.2024.1355460

In vitro anti- Helicobacter pylori activity and the underlining mechanism of an empirical herbal formula - Hezi Qingyou

Front Microbiol. 2024 Feb 19:15:1355460. doi: 10.3389/fmicb.2024.1355460. eCollection 2024.

Authors

Zhong Feng^#^{1

2

3}, Hui Li^#^{2

3}, Yajie Hao^{2

3}, Chang Peng¹, Ling Ou¹, Junwei Jia^{2

3}, Mingjin Xun^{2

3}, Yuanjing Zou¹, Meiyun Chen¹, Guimin Zhang^{2

3}, Meicun Yao^{1

4}

Affiliations

¹ School of Pharmaceutical Sciences, Sun Yat-sen University, Shenzhen, China.
² State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine, Lunan Pharmaceutical Group Co., Ltd., Linyi, China.
³ International Pharmaceutical Engineering Laboratory in Shandong Province, Shandong New Time Pharmaceutical Co., Ltd., Linyi, China.
⁴ Nanchang Research Institute, Sun Yat-sen University, Jiangxi, China.

^# Contributed equally.

Abstract

Background: Helicobacter pylori (H. pylori) is thought to primarily colonize the human stomach and lead to various gastrointestinal disorders, such as gastritis and gastric cancer. Currently, main eradication treatment is triple or quadruple therapy centered on antibiotics. Due to antibiotic resistance, the eradication rate of H. pylori is decreasing gradually. Therefore, searching for anti-H. pylori drugs from herbal sources has become a strategy for the treatment. Our team proposed a Hezi Qingyou Formula (HZQYF), composed of Chebulae Fructus, Ficus hirta Vahl and Cloves, and studied its anti-H. pylori activity and mechanism.

Methods: Chemical components of HZQYF were studied using UHPLC-MS/MS and HPLC. Broth microdilution method and agar dilution method were used to evaluate HZQYF's antibacterial activity. The effects of HZQYF on expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF), and flagellar genes (flaA, flaB) were explored using Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) technology. Effects on morphology and permeability of the extracellular membrane were studied using scanning electron microscopy (SEM) and N-phenylnaphthalen-1-amine (NPN) uptake. Effect on urease activity was studied using a urease kinetics analysis in vitro. Immunofluorescence staining method was used to examine the effect on adhesion. Western blot was used to examine the effect on cagA protein.

Results: Minimum inhibitory concentration (MIC) values of the formula against H. pylori clinical strains and standard strains were 80-160 μg/mL, and minimum bactericidal concentration (MBC) values were 160-320 μg/mL. The formula could down-regulate the expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF) and flagellar genes (flaA, flaB), change the morphology of H. pylori, increase its extracellular membrane permeability, and decrease its urease activity.

Conclusion: Present studies confirmed that HZQYF had promising in vitro anti-H. pylori activities and demonstrated its possible mechanism of action by down-regulating the bacterial adhesion, urease, and flagellar gene expression, which provided scientific bases for further clinical investigations.

Keywords: Helicobacter pylori; Hezi Qingyou Formula; antibacterial activity; in vitro; mechanism of action.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work has been supported by the National Natural Science Foundation of China (grants 81973552) and the Key R&D Program of Shandong Province, China (2020CXGC010506).