Actinobacillus actinomycetemcomitans lipopolysaccharide induces interleukin-6 expression through multiple mitogen-activated protein kinase pathways in periodontal ligament fibroblasts

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorPatil, C.
dc.creatorRossa, C.
dc.creatorKirkwood, K. L.
dc.date2014-02-26T17:27:02Z
dc.date2014-05-20T13:45:06Z
dc.date2016-10-25T16:59:11Z
dc.date2014-02-26T17:27:02Z
dc.date2014-05-20T13:45:06Z
dc.date2016-10-25T16:59:11Z
dc.date2006-12-01
dc.date.accessioned2017-04-05T20:51:12Z
dc.date.available2017-04-05T20:51:12Z
dc.identifierOral Microbiology and Immunology. Oxford: Blackwell Publishing, v. 21, n. 6, p. 392-398, 2006.
dc.identifier0902-0055
dc.identifierhttp://hdl.handle.net/11449/15843
dc.identifierhttp://acervodigital.unesp.br/handle/11449/15843
dc.identifier10.1111/j.1399-302X.2006.00314.x
dc.identifierWOS:000241500600009
dc.identifierhttp://dx.doi.org/10.1111/j.1399-302X.2006.00314.x
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/862765
dc.descriptionActinobacillus actinomycetemcomitans plays a major role in the pathogenesis of aggressive periodontitis. Lipopolysaccharide (LPS) derived from A. actinomycetemcomitans is a key factor in inflammatory cytokine generation within periodontal tissues. In this study, we identify major mitogen-activated protein kinase (MAPK) signaling pathways induced by A. actinomycetemcomitans LPS, Escherichia coli LPS and interleukin-1 beta (IL-1 beta) in a murine periodontal ligament (mPDL) fibroblast cell line. Immunoblot analysis was used to assess the phosphorylated forms of p38, extracellular-regulated kinase (ERK) and c-jun N-terminal kinase (JNK) MAPK following stimulation with A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta. IL-6 mRNA induction was detected via reverse transcription-polymerase chain reaction, while protein levels were quantified via enzyme-linked immunosorbent assays (ELISA). We utilized biochemical inhibitors of p38, ERK and JNK MAPK to identify the MAPK signaling pathways needed for IL-6 expression. Additional use of stable mPDL cell lines containing dominant negative mutant constructs of MAPK kinase-3 and -6 (MKK-3/6) and p38 null mutant mouse embryonic fibroblast (MEF) cells were used to substantiate the biochemical inhibitor data. Blocking p38 MAPK with SB203580 reduced the induction of IL-6 mRNA by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by > 70%, > 95% and similar to 60%, respectively. IL-6 ELISA indicated that blocking p38 MAPK reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by similar to 60%, similar to 50% and similar to 70%, respectively. All MAPK inhibitors significantly reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta whereas only p38 inhibitors consistently reduced the A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta induction of IL-6 mRNA steady-state levels. The contribution of p38 MAPK LPS-induced IL-6 expression was confirmed using MKK-3/6 dominant negative stable mPDL cell lines. Wild-type and p38 alpha(-/-) MEF cells provided additional evidence to support the role of p38 alpha MAPK in A. actinomycetemcomitans LPS-stimulated IL-6. Our results indicate that induction of IL-6 by E. coli LPS, IL-1 beta and A. actinomycetemcomitans LPS requires signaling through MKK-3-p38 alpha ERK, JNK and p38 MAPK in mPDL cells.
dc.languageeng
dc.publisherBlackwell Publishing
dc.relationOral Microbiology and Immunology
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectActinobacillus actinomycetemcomitans
dc.subjectinterleukin-6
dc.subjectLPS
dc.subjectMAPkinases
dc.subjectperiodontal disease
dc.subjectperiodontal ligament fibroblast
dc.titleActinobacillus actinomycetemcomitans lipopolysaccharide induces interleukin-6 expression through multiple mitogen-activated protein kinase pathways in periodontal ligament fibroblasts
dc.typeOtro


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