PGE2 induces IL-1beta gene expression in mouse osteoblasts through a cAMP-PKA signaling pathway.
Prostaglandin E2 (PGE2), an abundant eicosanoid in bone, has been implicated in a number of pathological states associated with bone loss and is also known to stimulate matrix metalloproteinase-1 synthesis and secretion in rat and human osteoblast cells, although the intracellular reactions responsible for this remain unclear. Interleukin-1beta (IL-1beta) is a cytokine that plays a critical role in bone remodeling and appears to act as a downstream effector of most bone-resorbing agents. However, the issue of whether PGE2 regulates the expression of IL-1beta in mouse osteoblasts has not been resolved. In this work, we demonstrate that PGE2 is a potent inducer of IL-1beta production by fetal osteoblasts and show that PGE2 stimulates the activity of the IL-1beta promoter in osteoblasts, suggesting that PGE2 controls IL-1beta gene expression at least at the transcriptional level. PGE2 was found to induce IL-1beta mRNA expression in the cells within 4 h and the level of expression was maintained for 36 h. A dose-related increase in IL-1beta production was found with 0.1-2.0 microM PGE2. The induction of IL-1beta protein in the medium paralleled the induction of IL-1beta mRNA levels. The role of cAMP activation in PGE2-mediated IL-1beta production was examined by the effects of forskolin, an adenylyl cyclase (AC) activator and dideoxyadenosine (DDA), an AC inhibitor. Forskolin enhanced and DDA blocked the production of IL-1beta by PGE2. In addition, PGE2-mediated IL-1beta induction was completely inhibited by the cAMP antagonist, Rp-cAMP, and protein kinase A (PKA) inhibitors of KT5720 and H89. The PGE2-induced production of IL-1beta was also blocked by the PKA inhibitor PKI14-22. However, a specific inhibitor of protein kinase C, calphostin C, had no affect on PGE2-induced IL-1beta gene expression. Among the potential agonists, forskolin was a potent inducer of IL-1beta expression, while phorbol myristate acetate and serum had little effect. These findings indicate that PGE2 involves the cAMP-PKA signaling pathway in regulating IL-1beta gene expression in osteoblasts.
Park
Young-Guk
YG
Department of Orthodondritics, Kyung-Hee University College of Dental Medicine, Dongdaemun-Gu, Seoul 130-701, South Korea. ygpark@khu.ac.kr
Kang
Sung-Koo
SK
Noh
Sang-Ho
SH
Park
Kwan-Kyu
KK
Chang
Young-Chae
YC
Lee
Young-Choon
YC
Kim
Cheorl-Ho
CH
D000067956:Adenylyl Cyclase Inhibitors; D000818:Animals; D000831:Animals, Newborn; D015152:Blotting, Northern; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D016048:Dideoxyadenosine / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects*; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000067956:Adenylyl Cyclase Inhibitors; D004791:Enzyme Inhibitors; D007375:Interleukin-1; D012333:RNA, Messenger; D005576:Colforsin; D016048:Dideoxyadenosine; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate