Abstract / Summary
Background/Objectives: HF33 is a polyherbal formulation developed from Thai folk medicinal knowledge for the management of dyspepsia and gastric ulcer-like symptoms. Its gastroprotective activity and underlying mechanisms have not been systematically investigated. This study aimed to characterize the HF33 formulation and evaluate the gastroprotective activity, possible mechanisms of action, and acute oral toxicity of its ethanolic extract. Methods: HF33 formulation powder was characterized according to the Thai Herbal Pharmacopoeia using pharmacognostic, physicochemical, and HPLC-PDA analyses, and the extract was chemically fingerprinted by compact mass spectrometry. Antioxidant activity and total phenolic and flavonoid contents were determined. Gastroprotective activity of the extract (150, 300, and 600 mg/kg, p.o.) was evaluated in male Sprague–Dawley rats using restraint water immersion stress-, indomethacin-, and acidified ethanol (EtOH/HCl)-induced gastric ulcer models. Histopathology, MDA and SOD, gastric mucus content, and gastric secretion parameters were assessed. Acute oral toxicity was evaluated in female Sprague–Dawley rats according to OECD Test Guideline 420. Results: Quality characterization established the identity and quality attributes of the HF33 formulation and its extract prior to biological evaluation. The HF33 extract exhibited in vitro antioxidant activity and significantly reduced gastric lesion formation in all three ulcer models. Gastroprotection was associated with preservation of gastric mucosal architecture, increased gastric mucus content, reduced MDA levels, and restored SOD activity, without significant alteration of gastric secretion parameters. No treatment-related toxicity was observed following a single oral administration of 2000 mg/kg. Conclusions: The HF33 ethanolic extract exhibited gastroprotective activity across multiple experimental gastric ulcer models. The protective activity was associated with preservation of mucosal integrity, enhancement of the mucus barrier, and attenuation of oxidative damage without affecting gastric acid secretion. These findings support further preclinical investigation of HF33 as a quality-characterized polyherbal formulation with potential gastroprotective properties.