Correlates of MAPK/Nrf2/STAT‐3 Pathway Protein Expression With Morin Treatment in Helicobacter pylori ‐Infected Gastric Epithelial Cells‐1 Cells: An Exploratory In Vitro Study

Helicobacter pylori infection is a major risk factor for gastric cancer (GC), but the molecular mechanisms remain incompletely understood. This exploratory in vitro study investigated whether treatment with the flavonoid morin correlates with altered expression of MAPK/Nrf2/STAT-3 pathway proteins in H. pylori-infected human gastric epithelial cells (GES-1). GES-1 cells were infected with H. pylori (ATCC 49503, MOI 100:1) and treated with 40 µM morin for 24 h. Outcomes included cell viability (MTT assay), ROS production (DCFH-DA), oxidative DNA damage (comet assay), glutathione and malondialdehyde levels (biochemical assays), protein expression (Western blot), and gene expression (RT-PCR). Molecular docking predicted binding affinity between morin and pathway proteins. This study used n = 3 biological replicates per group and a lenient false discovery rate threshold (Q = 0.25) appropriate for hypothesis generation; findings require independent replication. In this exploratory study, morin treatment (40 µM) was associated with: Higher MTT-detectable viability in H. pylori-infected cultures compared to infected untreated controls (85% vs. 45%; q = 0.042) Lower intracellular ROS levels (q = 0.021) and reduced oxidative DNA damage (q = 0.018) compared to infected untreated controls Higher GSH and lower MDA levels compared to infected untreated controls Lower phosphorylation levels of MAPK family members (ERK, JNK, p38), PI3K/AKT, and STAT3/EGFR proteins compared to infected untreated controls Higher total Nrf2 protein expression and upregulation of HMOX1 and NQO1 mRNA compared to infected untreated controls Molecular docking predicted binding between morin and MAPK, STAT3, and NRF2 pathway proteins, but these computational findings require experimental validation. These correlative findings are consistent with the hypothesis that morin treatment engages MAPK/Nrf2/STAT-3 pathways in H. pylori-infected GES-1 cells. However, this study does not establish causality, Nrf2 activation (nuclear translocation not demonstrated), or therapeutic efficacy (no positive controls). The observed associations should be interpreted as hypothesis-generating and require independent replication, mechanistic validation (including Nrf2 loss-of-function experiments), and comparative studies with standard agents before any translational inference. Under the specific in vitro conditions tested (GES-1 cells, H. pylori ATCC 49503, 40 µM morin, 24-h exposure, n = 3 biological replicates), morin treatment was associated with lower ROS levels, reduced MAPK/STAT3 phosphorylation, and higher Nrf2 protein expression compared to infected untreated controls. These correlative findings are hypothesis-generating and do not establish therapeutic potential, safety, efficacy, or clinical relevance. Independent replication, comprehensive toxicological characterization (including normal cell lines and in vivo models), direct comparison with standard agents (clarithromycin, sulforaphane), and mechanistic validation (including Nrf2 loss-of-function experiments) are required before any consideration of morin for further development. This study provides no evidence to support morin as a therapeutic, adjuvant, or alternative agent.