JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
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Regulatory effects of interleukin-1beta and prostaglandin E2 on expression of receptor activator of nuclear factor-kappaB ligand in human periodontal ligament cells.

BACKGROUND: Receptor activator of nuclear factor-kappaB ligand (RANKL), which is expressed on the cell membrane of osteoblasts/stromal cells, stimulates osteoclastogenesis. We investigated the regulatory effects of interleukin-1beta (IL-1beta) and prostaglandin E2 (PGE2) on expression of RANKL in human periodontal ligament (HPDL) cells and the mechanisms involved in the PGE2 effect.

METHODS: The HPDL cells were treated with IL-1beta, alone or in combination with indomethacin (INDO) or NS398, a cyclooxygenase-2 (COX-2) inhibitor. The HPDL cells were also pretreated with H89, a protein kinase A (PKA) inhibitor or GF109203X, a protein kinase C (PKC) inhibitor and subsequently treated with PGE2, PGE receptor (EP)2 agonist, EP4 agonist, forskolin, dibutyryl cAMP (db-cAMP), or 3-(isobutyl)-1-methylxantine (IBMX). After each treatment, expression of EP2, EP4, or RANKL mRNA was analyzed by reverse transcription-polymerase chain reaction and Southern hybridization. Expression of RANKL protein was detected by Western blotting, and cAMP accumulation was determined using a cAMP enzyme immunoassay kit.

RESULTS: IL-1beta stimulated the expression of RANKL at messenger RNA (mRNA) and protein levels in HPDL cells. Endogenous PGE2 partially mediated the IL-1beta-induced RANKL mRNA expression. Exogenously added PGE2 also stimulated RANKL expression at mRNA and protein levels in the cells. The PGE2-stimulated RANKL expression was mediated by EP2/4 and cAMP-dependent PKA, while PKC was possibly involved in the PGE2 action.

CONCLUSION: Human periodontal ligament cells activated with inflammatory factors such as IL-1beta and PGE2 may directly stimulate osteoclastogenesis through RANKL, which is stimulated to express by these factors.

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