Abstract
Background: Helicobacter pylori is a widespread gastric pathogen associated with chronic gastritis, peptic ulcer disease, and gastric adenocarcinoma. The increasing antibiotic resistance and treatment failure with standard triple therapy necessitate the exploration of alternative or adjunctive therapeutic agents with immunomodulatory properties.the objective of this study aimed to evaluate the therapeutic efficacy of bovine lactoferrin (bLf) alone and in combination with standard triple therapy (TT) (Amoxicillin (AMX), Clarithromycin (CLR), and proton pump inhibitors PPIs) against H. pylori infection in a BALB/c mouse model, with assessment of bacterial clearance and suppression of virulence factor gene expression. The methods found sixty BALB/c mice were randomly divided into five groups (n=12/group). Groups 1-4 were infected intragastrically with H. pylori clinical isolate (1×10⁹ CFU/mL), while Group 5 served as uninfected negative control. One-week post-infection, two-week daily treatment was administered as follows: Group 1 (positive control, infected untreated), Group 2 (triple therapy alone), Group 3 (bovine lactoferrin alone, 100 mg/kg), Group 4 (triple therapy plus bovine lactoferrin), and Group 5 (negative control, PBS only). Gastric tissue samples were collected for bacterial colony counting and quantitative real-time PCR analysis of cagA and vacA virulence gene expression. The results found a bacterial load was significantly reduced in all treatment groups compared to positive control (P < 0.0001). The greatest reduction was observed in Group 4 (triple therapy plus bLf; 5.457 ± 0.024 log₁₀ CFU/mL), representing an 86.9% reduction compared to Group 1 (6.340 ± 0.015 log₁₀ CFU/mL). Gene expression analysis revealed significant suppression of cagA and vacA in treated groups, with maximal suppression in Group 4 (83.5% reduction for cagA; 79.0% reduction for vacA; P < 0.05). In conclusion the study found combination therapy comprising standard triple therapy and bovine lactoferrin demonstrated superior antibacterial efficacy and virulence gene suppression compared to either treatment alone. These findings suggest that bovine lactoferrin may serve as an effective adjunctive agent in H. pylori eradication therapy, potentially through both direct antimicrobial and immunomodulatory mechanisms.