Evaluation of Helicobacter pylori-derived outer membrane vesicles on the expression of inflammatory cytokines

Maryam Mohammadbeigi; Amir Peymani; Shahin Bolori; Simin Sotoudeh; Rasoul Samimi; Anahita Bakhtiari; Saina Shegefti; Mehdi Bakht; Milad Badri; Amir Javadi; Farhad Nikkhahi

doi:10.18502/ijm.v17i6.20365

Maryam Mohammadbeigi Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
Amir Peymani Department of Microbiology, Mazandaran University of Medical Sciences, Ramsar, Iran
Shahin Bolori Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
Simin Sotoudeh Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
Rasoul Samimi Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
Anahita Bakhtiari Department of Microbiology, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Saina Shegefti Department of Microbiology, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Mehdi Bakht Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
Milad Badri Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
Amir Javadi Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
Farhad Nikkhahi Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran

DOI: https://doi.org/10.18502/ijm.v17i6.20365

Keywords: Helicobacter pylori; Gene expression; Gastric cancer; Real-time polymerase chain reaction; Virulence factor

Abstract

Background and Objectives: Helicobacter pylori infection has been increasingly linked to extra-gastric diseases. Outer membrane vesicles are a key virulence factor of H. pylori. This study investigates the influence of H. pylori-derived outer membrane vesicles on inflammatory marker expression in human hepatoma cells (HepG2).

Materials and Methods: Outer membrane vesicles were isolated through ultracentrifugation and characterized using dy- namic light scattering technique (DLS) and a Field Emission Scanning Electron Microscope (FE-SEM). Protein concen- trations were measured via the Bradford assay. HepG2 cells treated with outer membrane vesicles were analyzed for IL-6, TNF-α, TLR-4, TGF-β, and PPAR-γ mRNA expression by RT-qPCR. Cell viability was assessed through an MTT assay. The prevalence of H. pylori virulence-associated genes (babA2, sabA, and oipA) was determined by PCR.

Results: The results showed a high prevalence of sabA (91.7%), babA2 (75%), and oipA (66.7%). FE-SEM and DLS anal- yses confirmed the presence of bleb-shaped nanovesicles ranging in size from 50 to 450 nm. H. pylori-derived outer mem- brane vesicles significantly upregulated the expression of pro-inflammatory markers (TLR-4, PPAR-γ, TNF-α, and IL-6), while downregulating TGF-β expression.

Conclusion: These findings underscore the potential role of nanoparticles in driving inflammatory responses and influencing host cell signaling, which may play a key role in liver-related pathologies.