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.
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.
Authors: B S Herrera; T Ohira; L Gao; K Omori; R Yang; M Zhu; M N Muscara; C N Serhan; T E Van Dyke; R Gyurko Journal: Br J Pharmacol Date: 2008-09-22 Impact factor: 8.739
Authors: M Kiji; T Nagasawa; D Hormdee; R Yashiro; H Kobayashi; K Noguchi; H Nitta; Y Izumi; I Ishikawa Journal: Clin Exp Immunol Date: 2007-06-05 Impact factor: 4.330