Nuclear Factor-κB Dependent Induction of TNF-α and IL-1β by the Aggregatibacter actinomycetemcomitans Lipopolysaccharide in RAW 264.7 Cells
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초록
Aggregatibacter actinomycetemcomitans is an importantpathogen in the development of localized aggressiveperiodontitis. Lipopolysaccharide (LPS) is a virulent factorof periodontal pathogens that contributes to alveolar boneloss and connective tissue degradation in periodontaldisease. Our present study was designed to investigate thecytokine expression and signaling pathways regulated by A. actinomycetemcomitans LPS (Aa LPS). Cytokine geneexpression profiling in RAW 264.7 cells was performed bymicroarray analyses. The cytokine mRNA and proteinlevels and related signaling pathways induced by Aa LPSwere measured by RT-PCR, ELISA and western blotting. Microarray results showed that Aa LPS strongly inducedthe expression of NF-κB, NF-κB-related genes, inflammatorycytokines, TNF-α and IL-1β in RAW 264.7 cells. NF-κBinhibitor pretreatment significantly reduced the levels ofTNF-α and IL-1β mRNA and protein. In addition, the AaLPS-induced TNF-α and IL-1β expression was inhibited byp38/JNK MAP kinase inhibitor pretreatment. These resultsshow that Aa LPS stimulates TNF-α and IL-1β expressionthrough NF-κB and p38/JNK activation in RAW 264.7cells, suggesting the essential role of this pathway in the pathogenesis of localized aggressive periodontitis.
Abstract
Aggregatibacter actinomycetemcomitans is an importantpathogen in the development of localized aggressiveperiodontitis. Lipopolysaccharide (LPS) is a virulent factorof periodontal pathogens that contributes to alveolar boneloss and connective tissue degradation in periodontaldisease. Our present study was designed to investigate thecytokine expression and signaling pathways regulated by A. actinomycetemcomitans LPS (Aa LPS). Cytokine geneexpression profiling in RAW 264.7 cells was performed bymicroarray analyses. The cytokine mRNA and proteinlevels and related signaling pathways induced by Aa LPSwere measured by RT-PCR, ELISA and western blotting. Microarray results showed that Aa LPS strongly inducedthe expression of NF-κB, NF-κB-related genes, inflammatorycytokines, TNF-α and IL-1β in RAW 264.7 cells. NF-κBinhibitor pretreatment significantly reduced the levels ofTNF-α and IL-1β mRNA and protein. In addition, the AaLPS-induced TNF-α and IL-1β expression was inhibited byp38/JNK MAP kinase inhibitor pretreatment. These resultsshow that Aa LPS stimulates TNF-α and IL-1β expressionthrough NF-κB and p38/JNK activation in RAW 264.7cells, suggesting the essential role of this pathway in the pathogenesis of localized aggressive periodontitis.
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