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Inhibitors of mitogen-activated protein kinases downregulate COX-2 expression in human chondrocytes
Riina Nieminen1, Sari Leinonen, Aleksi Lahti
1The Immunopharmacology Research Group, Medical School, University of Tampere, and Tampere University Hospital, Research Unit, Finland.
Abstract:
Inducible prostaglandin synthase (cyclooxygenase-2, COX-2) is expressed in rheumatoid and osteoarthritic cartilage and produces high amounts of proinflammatory prostanoids in the joint. In the present study we investigated the effects of the inhibitors of mitogen-activated protein kinase (MAPK) pathways Erk1/2, p38, and JNK on COX-2 expression and prostaglandin E2 (PGE2) production in human chondrocytes. Proinflammatory cytokine IL-1beta caused a transient activation of Erk1/2, p38, and JNK in immortalized human T/C28a2 chondrocytes and that was followed by enhanced COX-2 expression and PGE2 production. PD98059 (an inhibitor of Erk1/2 pathway) suppressed IL-1-induced COX-2 expression and PGE2 production in a dose-dependent manner, and seemed to have an inhibitory effect on COX-2 activity. SB203580 (an inhibitor of p38 pathway) but not its negative control compound SB202474 inhibited COX-2 protein and mRNA expression and subsequent PGE2 synthesis at micromolar drug concentrations. SP600125 (a recently developed JNK inhibitor) but not its negative control compound N1-methyl-1,9-pyrazolanthrone downregulated COX-2 expression and PGE2 formation in a dose-dependent manner. SP600125 did not downregulate IL-1-induced COX-2 mRNA expression when measured 2 h after addition of IL-1beta but suppressed mRNA levels in the later time points suggesting post-transcriptional regulation. Our results suggest that activation of Erk1/2, p38, and JNK pathways belongs to the signaling cascades that mediate the upregulation of COX-2 expression and PGE2 production in human chondrocytes exposed to proinflammatory cytokine IL-1beta.
Insights
Mitogen-activated protein kinase (MAPK) pathways Erk1/2, p38, and JNK activation increases cyclooxygenase-2 (COX-2) expression and prostaglandin E2 (PGE2) production in human chondrocytes. Inhibiting these pathways reduces inflammation markers in joint cells.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Rheumatoid and osteoarthritis involve cyclooxygenase-2 (COX-2) expression in cartilage, leading to pro-inflammatory prostanoid production.
- Understanding the regulation of COX-2 in chondrocytes is crucial for developing targeted therapies for joint inflammation.
Purpose of the Study:
- To investigate the impact of mitogen-activated protein kinase (MAPK) pathway inhibitors (Erk1/2, p38, JNK) on COX-2 expression and prostaglandin E2 (PGE2) production in human chondrocytes.
- To elucidate the role of specific MAPK pathways in mediating IL-1beta-induced COX-2 upregulation.
Main Methods:
- Utilized immortalized human T/C28a2 chondrocytes.
- Administered IL-1beta to induce inflammation and subsequently applied specific MAPK pathway inhibitors (PD98059 for Erk1/2, SB203580 for p38, SP600125 for JNK).
- Assessed COX-2 expression (protein and mRNA) and PGE2 production using various concentrations of inhibitors and control compounds; analyzed temporal effects on mRNA expression.
Main Results:
- IL-1beta induced transient activation of Erk1/2, p38, and JNK pathways, followed by increased COX-2 expression and PGE2 production.
- PD98059 (Erk1/2 inhibitor) dose-dependently suppressed IL-1beta-induced COX-2 expression and PGE2 production.
- SB203580 (p38 inhibitor) inhibited COX-2 protein and mRNA expression and PGE2 synthesis at micromolar concentrations.
- SP600125 (JNK inhibitor) dose-dependently reduced COX-2 expression and PGE2 formation, with evidence of post-transcriptional regulation impacting later mRNA levels.
Conclusions:
- Activation of Erk1/2, p38, and JNK pathways are key signaling cascades involved in the IL-1beta-induced upregulation of COX-2 expression and PGE2 production in human chondrocytes.
- Targeting these MAPK pathways offers a potential therapeutic strategy for managing inflammatory conditions in joints characterized by elevated COX-2 activity.
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