Periodontopathogen LPSs Regulate MicroRNA Expression in Human Gingival Epithelial Cells
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초록
Periodontitis results from the activation of host immune and inflammatory defense responses to subgingival plaque bacteria, most of which are gram-negative rods with lipopolysaccharides (LPSs) in their cell walls. LPSs have been known to induce proinflammatory responses and recently it was reported also that they induce the expression of microRNAs (miRNAs) in host cells. In our current study therefore, we aimed to examine and compare the miRNA expression patterns induced by the LPSs of major periodontopathogens in the human gingival epithelial cell line, Ca9-22. The cells were treated with 1 μg/ml of E. coli (Ec) LPS or 5 μg/ml of an LPS preparations from four periodontopathogens Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn) for 24 h. After small RNA extraction from the treated cells, miRNA microarray analysis was carried out and characteristic expression profiles were observed. Fn LPS most actively induced miRNAs related to inflammation,followed by Aa LPS, Pi LPS, and Ec LPS. In contrast, Pg LPS only weakly activated miRNAs related to inflammation. Among the miRNAs induced by each LPS, miR-875-3p,miR-449b, and miR-520d-3p were found to be commonly up-regulated by all five LPS preparations, although at different levels. When we further compared the miRNA expression patterns induced by each LPS, Ec LPS and Pi LPS were the most similar although Fn LPS and Aa LPS also induced a similar miRNA expression pattern. In contrast,the miRNA profile induced by Pg LPS was quite distinctive compared with the other bacteria. In conclusion, miR-875-3p, miR-449b, and miR-520d-3p miRNAs are potential targets for the diagnosis and treatment of periodontal inflammation induced by subgingival plaque biofilms. Furthermore, the observations in our current study provide new insights into the inflammatory miRNA response to periodontitis.
Abstract
Periodontitis results from the activation of host immune and inflammatory defense responses to subgingival plaque bacteria, most of which are gram-negative rods with lipopolysaccharides (LPSs) in their cell walls. LPSs have been known to induce proinflammatory responses and recently it was reported also that they induce the expression of microRNAs (miRNAs) in host cells. In our current study therefore, we aimed to examine and compare the miRNA expression patterns induced by the LPSs of major periodontopathogens in the human gingival epithelial cell line, Ca9-22. The cells were treated with 1 μg/ml of E. coli (Ec) LPS or 5 μg/ml of an LPS preparations from four periodontopathogens Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Aggregatibacter actinomycetemcomitans (Aa), and Fusobacterium nucleatum (Fn) for 24 h. After small RNA extraction from the treated cells, miRNA microarray analysis was carried out and characteristic expression profiles were observed. Fn LPS most actively induced miRNAs related to inflammation,followed by Aa LPS, Pi LPS, and Ec LPS. In contrast, Pg LPS only weakly activated miRNAs related to inflammation.
참고문헌 (33)
- Bartel DP, "target recognition and regulatory functions", Cell, 136, pp.215~233 (2009)
- Birkedal-Hansen H, "Role of cytokines and inflammatory mediators in tissue destruction", J. Periodontal Res, 28, pp.500~510 (1993)
- Bramanti, T. E., "Chemical characterization and biologic properties of lipopolysaccharide from Bacteroides gingivalis strains W50, W83, and ATCC 33277", Oral Microbiol. Immunol, 4, pp.183~192 (1989)
- Chang TC, "Transactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosis", Mol Cell, 26, pp.745~752 (2009)
- Chiang CY, "Effects of CD14 receptors on tissue reactions induced by local injection of two Gramnegative bacterial lipopolysaccharides", J Periodontal Res, 38, pp.36~43 (2003)
- Diya Zhang, "Lipopolysaccharide (LPS) of Porphyromonas gingivalis induces IL-1beta, TNF-alpha and IL-6 production by THP-1 cells in a way different from that of Escherichia coli LPS", Innate Immun, 14(2), pp.99~107 (2008)
- Fujiwara T, "Serological properties and immunobiological activities of lipopolysaccharides from black-pigmented and related oral Bacteroides species", J Gen Microbiol, 134, pp.2867~2876 (1988)
- Garrison SW, "Lipopolysaccharide-stimulated PGE2 release from human monocytes. Comparison of lipopolysaccharides prepared from suspected periodontal pathogens", J Periodontol, 59, pp.684~687 (1988)
- Gemmell, E., "Cytokines and prostaglandins in immune homeostasis and tissue destruction in periodontal disease", Periodontol, 14, pp.112~143 (1997)
- Guarnieri DJ, "a new class of gene regulators", Ann Med, 40, pp.197~208 (2008)
- Haffajee AD, "Microbial etiological agents of destructive periodontal diseases", Periodontol, 5, pp.78~111 (1994)
- Hashimoto M, "Chemical structure and immunobiological activity of lipid A from Prevotella intermedia ATCC 25611 lipopolysaccharide", FEBS Lett, 543(1), pp.98~102 (2003)
- Hofstad, T., "Chemical composition, serologic reactivity and endotoxicity of lipopolysaccharides extracted in different ways from Bacteroides fragilis, Bacteroides melaninogenicus and Bacteroides oralis", Acta Pathol. Microbiol. Scand. Sect. B, 85, pp.262~270 (1977)
- Jotwani R, "Nuclear factorkappaB p50 subunits in chronic periodontitis and Porphyromonas gingivalis lipopolysaccharide-pulsed dendritic cells", Ann N Y Acad Sci, 1192, pp.278~275 (2010)
- Koga, T., "Biochemical and immunobiological properties of lipopolysaccharide (LPS) from Bacteroides gingivalis and comparison with LPS from Escherichia coli", Infect. Immun, 47, pp.638~647 (1985)
- Koga T, "Biochemical and immunobiological properties of lipopolysaccharide (LPS) from Bacteroides gingivalis and comparison with LPS from Escherichia coli", Infect Immun, 47(3), pp.638~647 (1985)
- Kumada H, "Structural study on the free lipid A isolated from lipopolysaccharide of Porphyromonas gingivalis", J Bacteriol, 177(8), pp.2098~2106 (1995)
- Kumada, H, "Structural study on the free lipid A isolated from lipopolysaccharide of Porphyromonas gingivalis", J. Bacteriol, 177, pp.2098~2106 (1995)
- Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method", Methods, 25, pp.402~408 (2001)
- Mansheim, B. J, "Immunochemical and biologic studies of the lipopolysaccharide of Bacteroides melaninogenicius subspecies asaccharolyticus", J. Immunol, 120, pp.72~78 (1978)
- Matsushima K, "MicroRNA signatures in Helicobacter pylori-infected gastric mucosa", Int J Cancer, 128(2), pp.361~370 (2011)
- Nair, B. C, "Biological effects of a purified lipopolysaccharide from Bacteroides gingivalis", J. Periodontal Res, 18, pp.40~49 (1983)
- Oglesby IK, "MicroRNAs in inflammatory lung disease--master regulators or target practice", Respir Res, 28, pp.11~148 (2010)
- Perry MM, "Rapid changes in microRNA-146a expression negatively regulate the IL-1beta-induced inflammatory response in human lung alveolar epithelial cells", J. Immunol, 180, pp.5689~5698 (2008)
- Shenouda SK, "MicroRNA function in cancer: oncogene or a tumor suppressor", Cancer Metastasis Rev, 28, pp.369~378 (2009)
- Simon BL, "The role of endotoxin in periodontal disease. II. Correlation of the amount of endotoxin in human gingival exudate with the clinical degree of inflammation", J Periodontol, 41, pp.81~86 (1970)
- Sun Y, "Gram-negative periodontal bacteria induce the activation of Toll-like receptors 2 and 4, and cytokine production in human periodontal ligament cells", J Periodontol, 81(10), pp.1488~1496 (2010)
- Taganov KD, "NF-B-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc", Natl. Acad. Sci. U S. A, 103, pp.12481~12486 (2006)
- Tobias, P. S., "Lipopolysaccharide dependent cellular activation", J. Periodontal Res, 32, pp.99~103 (1997)
- van Winkelhoff AJ, "Porphyromonas gingivalis, Bacteroides forsythus and other putative periodontal pathogens in subjects with and without periodontal destruction", J Clin Periodontol, 29, pp.1023~1028 (2002)
- Westphal, O, "Bacterial lipopolysaccharides: extraction with phenol water and further application of the procedure", Methods Carbohydr. Chem. R, 83 (1965)
- Yang X, "miR-449a and miR-449b are direct transcriptional targets of E2F1 and negatively regulate pRb-E2F1 activity through a feedback loop by targeting CDK6 and CDC25A", Genes Dev, 23(20), pp.2388~2393 (2009)
- Yoshimura A, "Secretion of IL-1 beta, TNF-alpha, IL-8 and IL-1ra by human polymorphonuclear leukocytes in response to lipopolysaccharides from periodontopathic bacteria", J Periodontal Res, 32(3), pp.279~286 (1997)