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Inducing Apical Periodontitis in Mice
Published on: August 6, 2019
Elevated MicroRNA-128 in Periodontitis Mitigates Tumor Necrosis Factor-α Response via p38 Signaling Pathway in
Hee Sam Na1, Mi Hee Park1, Yu Ri Song1
1Department of Oral Microbiology, School of Dentistry, Pusan National University, Yangsan, Korea.
Background:
Periodontitis is a chronic inflammatory disease resulting from an inflammatory response to subgingival plaque bacteria, including Porphyromonas gingivalis. MicroRNA (miRNA) is a current focus in regulating the inflammatory processes. In this study, the inflammatory miRNA expression in gingival tissues of patients with periodontitis and of healthy individuals is compared, and its role in regulating the inflammatory response is examined.
Methods:
Gingival tissues from patients with periodontitis and healthy individuals were collected for miRNA microarray. THP-1 and CA9-22 cells were challenged with P. gingivalis, and miRNA expression was determined by real-time polymerase chain reaction. Target genes for miRNA were predicted using TargetScanHuman database, and miRNA gene expressions were reviewed using public databases. For the functional study, THP-1 cells were transfected with a miRNA-128 mimic, and target gene expression was compared with THP-1 cells challenged with P. gingivalis. For the tolerance test, THP-1 cells transfected with miRNA-128 mimic were treated with phorbol 12-myristate 13-acetate (PMA) or paraformaldehyde (PFA)-fixed Escherichia coli. Tumor necrosis factor (TNF)-α production was determined by enzyme-linked immunosorbent assay, and mitogen-activated protein kinase (MAPK) protein phosphorylation was determined by Western blot.
Results:
Gingival tissues from patients with periodontitis showed increased expression of miRNA-128, miRNA-34a, and miRNA-381 and decreased expression of miRNA-15b, miRNA-211, miRNA-372, and miRNA-656. THP-1 cells and CA9-22 cells challenged with P. gingivalis showed increased miRNA-128 expression. Among the predicted miRNA-128 target genes, several genes that are involved in MAPK signaling pathway showed similar gene expression pattern between P. gingivalis challenge and miRNA-128 mimic transfection. In THP-1 cells transfected with miRNA-128 mimic, TNF-α production was lower, and phosphorylation of p38 was inhibited when challenged with PMA or PFA-fixed E. coli.
Conclusion:
miRNA-128 may be involved in mitigating the inflammatory response induced by P. gingivalis in periodontitis.
Insights
MicroRNA-128 may reduce inflammation in periodontitis. This study found increased miRNA-128 in patients and showed it lowers inflammatory responses to Porphyromonas gingivalis in cell models.
Area of Science:
- Oral biology
- Molecular biology
- Immunology
Background:
- Periodontitis is a chronic inflammatory disease linked to oral bacteria like Porphyromonas gingivalis.
- MicroRNAs (miRNAs) are key regulators of inflammatory processes, making them a focus for research.
Purpose of the Study:
- To compare miRNA expression in gingival tissues of periodontitis patients and healthy individuals.
- To investigate the role of specific miRNAs, particularly miRNA-128, in regulating inflammatory responses to P. gingivalis.
Main Methods:
- miRNA microarray analysis of gingival tissues.
- Real-time PCR to measure miRNA expression in cell lines stimulated with P. gingivalis.
- In vitro functional studies using miRNA mimics and Western blot to assess inflammatory pathways (MAPK) and cytokine production (TNF-α).
Main Results:
- Periodontitis tissues showed altered expression of several miRNAs, including increased miRNA-128.
- P. gingivalis challenge upregulated miRNA-128 in THP-1 and CA9-22 cells.
- miRNA-128 mimic transfection reduced TNF-α production and inhibited p38 MAPK phosphorylation in response to inflammatory stimuli.
Conclusions:
- miRNA-128 expression is dysregulated in periodontitis.
- miRNA-128 plays a role in mitigating the inflammatory response to P. gingivalis.
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