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Updated: Jan 13, 2026
Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity
Published on: March 31, 2021
Evaluation of Helicobacter pylori-derived outer membrane vesicles on the expression of inflammatory cytokines
Maryam Mohammadbeigi1, Amir Peymani2, Shahin Bolori1
1Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
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 dynamic light scattering technique (DLS) and a Field Emission Scanning Electron Microscope (FE-SEM). Protein concentrations 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 analyses confirmed the presence of bleb-shaped nanovesicles ranging in size from 50 to 450 nm. H. pylori-derived outer membrane 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.
Insights
Helicobacter pylori outer membrane vesicles, nanoparticles from the bacteria, significantly increase inflammatory markers in liver cells. This suggests a role for these bacterial nanoparticles in liver disease development.
Area of Science:
- Microbiology
- Cell Biology
- Nanotechnology
Background:
- Helicobacter pylori infection is linked to diseases beyond the stomach.
- Outer membrane vesicles (OMVs) are critical virulence factors of H. pylori.
Purpose of the Study:
- To investigate the impact of H. pylori-derived OMVs on inflammatory marker expression in human hepatoma (HepG2) cells.
Main Methods:
- OMVs isolated via ultracentrifugation, characterized by DLS and FE-SEM.
- HepG2 cells treated with OMVs; gene expression (IL-6, TNF-α, TLR-4, TGF-β, PPAR-γ) analyzed by RT-qPCR.
- Cell viability assessed by MTT assay; virulence genes (babA2, sabA, oipA) detected by PCR.
Main Results:
- High prevalence of H. pylori virulence genes: sabA (91.7%), babA2 (75%), oipA (66.7%).
- FE-SEM/DLS confirmed nanovesicle presence (50-450 nm).
- H. pylori OMVs upregulated pro-inflammatory markers (TLR-4, PPAR-γ, TNF-α, IL-6) and downregulated TGF-β.
Conclusions:
- H. pylori OMVs act as nanoparticles that drive inflammatory responses in liver cells.
- These bacterial nanoparticles may contribute to the pathogenesis of liver-related diseases.
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