1,3,6-Trigalloylglucose is a natural compound that can be extracted from the aqueous extracts of ripe fruit of Terminalia chebula Retz, commonly known as “Haritaki”. The potential anti-Helicobacter pylori (HP) activity of this compound has not been extensively studied or confirmed in scientific research. This compound was isolated using a semi-preparative liquid chromatography (LC) system and identified through Ultra-high-performance liquid chromatography–MS/MS (UPLC-MS/MS) and Nuclear Magnetic Resonance (NMR). Its role was evaluated using Minimum inhibitory concentration (MIC) assay and minimum bactericidal concentration (MBC) assay, scanning electron microscope (SEM), inhibiting kinetics curves, urea fast test, Cell Counting Kit-8 (CCK-8) assay, Western blot, and Griess Reagent System. Results showed that this compound effectively inhibits the growth of HP strain ATCC 700392, damages the HP structure, and suppresses the Cytotoxin-associated gene A (Cag A) protein, a crucial factor in HP infection. Importantly, it exhibits selective antimicrobial activity without impacting normal epithelial cells GES-1. In vitro studies have revealed that 1,3,6-Trigalloylglucose acts as an anti-adhesive agent, disrupting the adhesion of HP to host cells, a critical step in HP infection. These findings underscore the potential of 1,3,6-Trigalloylglucose as a targeted therapeutic agent against HP infections.

Shouhui Tongbian capsule (SHTB), a commercial Chinese patent medicine, has been shown to modulate gut microbiota dysbiosis and contains antibacterial active ingredients against Helicobacter pylori (H. pylori). H. pylori is a Gram-negative bacterium that colonizes the gastric mucosal and disrupt gastrointestinal microbiota balances. However, the precise mechanism underlying the anti-H. pylori effects of SHTB remains unclear. To investigate the in vitro anti-H. pylori effects and underlying mechanisms of SHTB. High-performance liquid chromatography (HPLC), ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), and network pharmacology analyses were used to identify the active compounds and potential targets of SHTB associated with H. pylori infection. The anti-H. pylori activity of SHTB was determined using the broth dilution method. Mechanistic studies were conducted through techniques such as electron microscopy observation, immunofluorescence staining, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blotting analyses. A total of 42 compounds were identified in SHTB. Database screening yielded 365 target genes related to SHTB constituents and 1035 genes associated with H. pylori infection. In vitro assays demonstrated that the minimum inhibitory concentration (MIC) of SHTB against both susceptible and resistant H. pylori strains ranged from 160 to 320 μg/mL, with the minimum bactericidal concentration (MBC) being twice the MIC. No antagonistic effects were observed when SHTB was combined with clarithromycin, metronidazole, levofloxacin, or amoxicillin. SHTB disrupted H. pylori morphology, reduced urease activity, and downregulated virulence-associated gene expression. Cellular assay further indicated that SHTB enhanced the anti-adhesion capability of host cells. Furthermore, SHTB alleviated H. pylori infection-induced damage by inhibiting the abnormal activation of the PI3K/AKT, MAPK, and NF-κB signaling pathway. These findings provide a theoretical basis for elucidating the mechanism of SHTB against H. pylori infection and confirm its protective role in combating H. pylori infection through the regulation of the PI3K/AKT, MAPK, and NF-κB signaling pathway network.