title: IL-17A stimulates the expression of inflammatory cytokines via celecoxib-blocked prostaglandin in MC3T3-E1 cells.
abstract: OBJECTIVE
  The prostaglandins (PGs) released from osteoblasts can alter the process of bone remodelling. Recently, we showed that compressive force induced the expression of pro-inflammatory cytokine interleukin (IL)-17s and their receptors in osteoblastic MC3T3-E1 cells and that IL-17A was expressed most highly. Consequently, in the current study we examined the effect of IL-17A and/or celecoxib on PGE(2) production and the expression of cyclooxygenases (COXs) and inflammatory cytokines in MC3T3-E1 cells. We also examined the effects of PGE(2) and cyclohexamide on the expression of inflammatory cytokines.
  
  
  METHODS
  Cells were cultured with or without IL-17A (0.1, 1.0, or 10 ng/ml) in the presence or absence of 10 microM celecoxib, a specific inhibitor of COX-2, for up to 72 h. Cells were pretreated with or without 10 microg/ml cycloheximide, protein synthesis inhibitor, for 30 min, and then cultured with 10 ng/ml IL-17A for 24 h. Cells were also cultured with or without 1.5 ng/ml PGE(2) for 24 h. PGE(2) production was determined by ELISA. The expression of COX-1, COX-2, IL-1alpha, IL-6, IL-8, IL-11, and TNF-alpha mRNAs and proteins was determined by real-time PCR and ELISA, respectively.
  
  
  RESULTS
  The expression of COX-2, IL-1alpha, IL-6, IL-8, IL-11, and TNF-alpha, as well as PGE(2) production increased in the presence of IL-17A, whereas COX-1 expression did not change. Celecoxib blocked the stimulatory effect of IL-17A on the expression of COX-2, IL-1alpha, IL-6, IL-8, and IL-11 as well as PGE(2) production, whereas it did not block TNF-alpha expression. Cycloheximide pretreatment suppressed the expression of IL-17-induced inflammatory cytokines. The expression of IL-1alpha, IL-6, IL-8, and IL-11 increased by the addition of PGE(2), whereas TNF-alpha expression was not affected.
  
  
  CONCLUSIONS
  These results suggest that IL-17A stimulates the expression of bone resorption-related inflammatory cytokines through an autocrine mechanism involving celecoxib-blocked PGs, mainly PGE(2), in osteoblasts.
authors.affiliation: Department of Orthodontics, Shandong University School of Dentistry, Jinan, Shandong Province, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.forename: Yuki
authors.initials: Y
authors.lastname: Koyama
authors.forename: Rina
authors.initials: R
authors.lastname: Sanuki
authors.forename: Narihiro
authors.initials: N
authors.lastname: Mitsui
authors.forename: Naoto
authors.initials: N
authors.lastname: Suzuki
authors.forename: Akemi
authors.initials: A
authors.lastname: Kimura
authors.forename: Akira
authors.initials: A
authors.lastname: Nakajima
authors.forename: Noriyoshi
authors.initials: N
authors.lastname: Shimizu
authors.forename: Masao
authors.initials: M
authors.lastname: Maeno
mesh_terms: D016475:3T3 Cells; D000818:Animals; D016723:Bone Remodeling / Q000187:drug effects / Q000502:physiology*; D000068579:Celecoxib; D003513:Cycloheximide / Q000494:pharmacology; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology*; D016207:Cytokines / Q000096:biosynthesis*; D003799:Dental Stress Analysis; D015232:Dinoprostone / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D018836:Inflammation Mediators / Q000378:metabolism; D020381:Interleukin-17 / Q000494:pharmacology / Q000502:physiology*; D007378:Interleukins / Q000096:biosynthesis; D051379:Mice; D010006:Osteoblasts / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D013449:Sulfonamides / Q000494:pharmacology
chemical_list: D052246:Cyclooxygenase 2 Inhibitors; D016207:Cytokines; D018836:Inflammation Mediators; D020381:Interleukin-17; D007378:Interleukins; D011500:Protein Synthesis Inhibitors; D011720:Pyrazoles; D012333:RNA, Messenger; D013449:Sulfonamides; D003513:Cycloheximide; D011451:Prostaglandin-Endoperoxide Synthases; D000068579:Celecoxib; D015232:Dinoprostone