Luteolin as a urease inhibitor: A novel therapeutic strategy against <i>Helicobacter pylori</i>

The rising prevalence of antibiotic-resistant Helicobacter pylori (H. pylori) underscores the urgent need for alternative treatment strategies. By producing ammonia to neutralize gastric acid, the key virulence factor urease is essential for H. pylori acid tolerance, thereby representing a promising therapeutic target. In this study, we identified luteolin as a potent urease inhibitor (IC50 = 30.50 μg/mL) from a screening of over 200 natural compounds. Further investigation through molecular docking, dynamics simulations, cellular thermal shift assay, and enzyme kinetics studies confirmed its competitive binding to the Ni2+-centered catalytic site of H. pylori urease (HPU). Luteolin exhibited potent anti-H. pylori activity under standard and simulated gastric conditions, and showed low propensity for resistance development over 14 serial passages. Proteomic and metabolomic analyses revealed that luteolin inhibited HPU activity, and the consequent ammonia restriction triggered severe metabolic dysfunction, characterized by disruptions in nucleotide, amino acid biosynthesis and TCA cycle. In GES-1 cells, luteolin protected against H. pylori-induced damage. In a mouse model of H. pylori-induced peptic ulcer, luteolin treatment significantly reduced the bacterial load, with concomitant alleviation of gastric mucosal pathology and suppression of inflammatory responses. In contrast to antibiotic-induced gastric microbial dysbiosis, microbial diversity analysis indicated that luteolin treatment had minimal impact on the resident gastric microbiota. In summary, as a urease inhibitor, luteolin suppresses H. pylori by blocking ammonia production, thereby disrupting acid neutralization and inducing metabolic dysfunction, which collectively alleviates gastric damage and inflammation while minimizing microbiota disruption and resistance risk.