Heme induces oxidative stress and inflammation in gastric epithelial cells via NADPH oxidase activation

Published: 16 March 2026| Version 1 | DOI: 10.17632/sr7jjpmd7w.1
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Description

Peptic ulcer disease is a prevalent condition that may give rise to several complications, including gastrointestinal bleeding, perforation, and gastric outlet obstruction. Vascular injury during ulceration promotes erythrocyte lysis, resulting in the release of elevated levels of free heme, which can directly interact with the gastric mucosa. Our group has previously demonstrated that heme acts as a pro-inflammatory and pro-oxidative molecule. In this context, oxidative stress has been identified as a central pathogenic mechanism in ulcer progression, leading to cellular dysfunction and tissue injury. In the present study, we investigated the effects of heme on gastric epithelial cell function.

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Human gastric epithelial cells (HGE3) were preincubated with diphenyleneiodonium (DPI) (10 µM), a selective NADPH oxidase inhibitor, and stimulated with Heme (10 µM). Reactive oxygen species (ROS) production (DCF and MitoSOX probes), protein expression (Western Blotting), and cytokine levels in the culture supernatant were analyzed (ELISA). The activity of nuclear factor kappa B (NFκB) (Luciferase Assay) and nitric oxide (NO) production (DAF probe) were also evaluated, as well as changes in cell permeability (Dextran-FITC extravasation). Male Swiss mice were pretreated with 100 μl via oral gavage with one of the following: DPI (10 µM), Heme (30 µM), or a combination of DPI and Heme at the same concentrations. One hour after pretreatment, gastric ulcers were induced by oral administration of 50% ethanol (0.5 mL/25 g).

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Pharmacology, Cell Biology, Inflammation, Oxidative Stress

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