Cartilage Biology
Effect of interleukin-1β on osteogenic protein 1–induced signaling in adult human articular chondrocytes
Amel M. Elshaier,
Arnavaz A. Hakimiyan,
Lev Rappoport,
David C. Rueger, Susan Chubinskaya,
Amel M. Elshaier
Rush University Medical Center, Chicago, Illinois
Search for more papers by this authorArnavaz A. Hakimiyan
Rush University Medical Center, Chicago, Illinois
Search for more papers by this authorLev Rappoport
Rush University Medical Center, Chicago, Illinois
Search for more papers by this authorCorresponding Author
Susan Chubinskaya
Rush University Medical Center, Chicago, Illinois
Department of Biochemistry, Cohn Research Building, Suite 522, Rush University Medical Center, 1735 West Harrison Street, Chicago, IL 60612Search for more papers by this authorAmel M. Elshaier,
Arnavaz A. Hakimiyan,
Lev Rappoport,
David C. Rueger, Susan Chubinskaya,
Amel M. Elshaier
Rush University Medical Center, Chicago, Illinois
Search for more papers by this authorArnavaz A. Hakimiyan
Rush University Medical Center, Chicago, Illinois
Search for more papers by this authorLev Rappoport
Rush University Medical Center, Chicago, Illinois
Search for more papers by this authorCorresponding Author
Susan Chubinskaya
Rush University Medical Center, Chicago, Illinois
Department of Biochemistry, Cohn Research Building, Suite 522, Rush University Medical Center, 1735 West Harrison Street, Chicago, IL 60612Search for more papers by this authorAbstract
Objective
Two major receptor-activated Smad (R-Smad) signaling pathways, bone morphogenetic protein (BMP) and MAPK, were examined in a model of interleukin-1β (IL-1β)–induced cartilage degeneration to investigate the effect of IL-1β on osteogenic protein 1 (OP-1) signaling in adult human articular chondrocytes.
Methods
Chondrocytes from the ankles of 26 normal human donors were cultured in high-density monolayers in serum-free medium. The effect of IL-1β on BMP receptors was studied by reverse transcription–polymerase chain reaction and flow cytometry. Phosphorylation of R-Smads was tested in cells treated with IL-1β (10 ng/ml), OP-1 (100 ng/ml), or the combination of IL-1β and OP-1. Cell lysates were analyzed by Western blotting with polyclonal antibodies against 2 R-Smad phosphorylation sites (BMP- and MAPK-mediated) or with total, nonphosphorylated R-Smad as a control. To identify which MAPKs play a role in IL-1β activation of the linker region, chondrocytes were preincubated with specific MAPK inhibitors (PD98059 for MAP/ERK, SP600125 for JNK, and SB203580 for p38).
Results
IL-1β reduced the number of activin receptor–like kinase 2 (ALK-2) and ALK-3 receptors, inhibited expression of Smad1 and Smad6, delayed and prematurely terminated the onset of OP-1–mediated R-Smad phosphorylation, and affected nuclear translocation of R-Smad/Smad4 complexes. The alternative phosphorylation of R-Smad in the linker region via the MAPK pathway (primarily p38 and JNK) was observed to be a possible mechanism through which IL-1β offsets OP-1 signaling and the response to OP-1. Conversely, OP-1 was found to directly inhibit phosphorylation of p38.
Conclusion
These findings describe new mechanisms of the crosstalk between OP-1 and IL-1β in chondrocytes. The study also identifies potential targets for therapeutic interventions in the treatment of cartilage-degenerative processes.
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