Amelioration of Helicobacter pylori-induced gastric injury by Artemisia argyi Folium total extract, its total flavonoids, and eupatilin through suppression of NF-κB signaling and virulence factors.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia argyi Folium has been used to treat gastrointestinal disorders like gastritis. Based on centuries-old folk practices, its aerial parts are made into decoctions, infusions, or moxibustion agents to relieve epigastric pain, reduce inflammation, and regulate digestive functions.

AIM OF THE STUDY: This study aimed to enrich and identify the flavonoid components, along with the active monomer eupatilin, from Artemisia argyi Folium. We aimed to compare their anti-inflammatory and antioxidant properties, assess their in vitro impact on Helicobacter pylori, and clarify their potential in vivo effects on the inhibition of virulence factors and NF-κB signaling activity.

MATERIALS AND

METHODS

The flavonoid profiles of two Artemisia argyi Folium extracts (AT and AF) were analyzed using UPLC-QE-MS. The antioxidant activities were evaluated in vitro using ABTS and DPPH assays; the anti-inflammatory effects were assessed employing RAW264.7 and GES-1 cell lines, and the MIC and MBC were determined. The therapeutic efficacy of AT, AF, and EUP against infection and gastric mucosal injury was investigated in H.pylori 43504-infected SD rat model. To probe the mechanisms of action, Western blotting and qPCR analyses were performed.

RESULTS

Comprehensive flavonoid profiling has revealed that enriched A. argyi flavonoids (AF, 86.6 %) possess significantly higher levels than total extracts (AT, 18.7 %) and demonstrate superior in vitro antioxidant activity, as evidenced by DPPH (IC50 = 133.8 ± 2.739 μg/ml) and ABTS (IC50 = 50.83 ± 2.071 μg/ml) assays, as well as anti-H. pylori activity (MIC = 320 μg/mL). The active monomer eupatilin (EUP) exhibits significant bactericidal activity (MIC = 16-32 μg/mL). In an H. pylori-infected SD rat model, all treatments (AT, AF, EUP) significantly reduced gastric bacterial load and ulcer area (e.g., EUP high-dose ulcer inhibition rate: 87.50 %, p < 0.001). They also markedly lowered plasma levels of virulence factors (CagA, VacA) and pro-inflammatory cytokines (IL-1β, IL-6, IL-8, TNF-α). Treatments also ameliorated gastric damage, enhanced antioxidant enzyme (SOD) activity, and reduced oxidative stress (MPO, MDA). Mechanistically, protection against H. pylori-induced inflammation and ulceration occurred via suppression of the NF-κB pathway.

CONCLUSIONS

This study provides in vivo and vitro confirmation that AT, AF, and EUP can effectively inhibit H. pylori in the gastric mucosa. By regulating the NF-κB signaling pathway and modulating virulence factors, it significantly reduces tissue damage in inflamed stomach areas, showing potential therapeutic efficacy for H. pylori-associated gastritis.