Abstract
This study was conducted to examine the cellular regulatory mechanisms of sulfasalazine (SSZ) in rabbit articular chondrocytes treated with sodium nitroprusside (SNP). Cell phenotype was determined, and the MTT assay, Western blot analysis and immunofluorescence staining of type II collagen was performed in control, SNP-treated and SNP plus SSZ (50~200 µg/mL) rabbit articular chondrocytes. Cellular proliferation was decreased significantly in the SNP-treated group compared with that in the control (p < 0.01). SSZ treatment clearly increased the SNP-reduced proliferation levels in a concentration-dependent manner (p < 0.01). SNP treatment induced significant dedifferentiation and inflammation compared with control chondrocytes (p < 0.01). Type II collagen expression levels increased in a concentration-dependent manner in response to SSZ treatment but were unaltered in SNP-treated chondrocytes (p < 0.05 and < 0.01, respectively). Cylooxygenase-2 (COX-2) expression increased in a concentration-dependent manner in response to SSZ treatment but was unaltered in SNP-treated chondrocytes (p < 0.05). Immunofluorescence staining showed that SSZ treatment increased type II collagen expression compared with that in SNP-treated chondrocytes. Furthermore, phosphorylated extracellular regulated kinase (pERK) expression levels were decreased significantly in the SNP-treated group compared with those in control chondro-cytes (p < 0.01). Expression levels of pERK increased in a concentration-dependent manner by SSZ but were unaltered in SNP-treated chondrocytes. pp38 kinase expression levels increased in a concentration-dependent manner by SSZ but were unaltered in control chondrocytes (p < 0.01). In summary, SSZ significantly inhibited nitric oxide-induced cell death and dedifferentiation, and regulated extracellular regulated kinases 1 and 2 and p38 kinase in rabbit articular chondrocytes.
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Abbreviations
- COX-2:
-
Cyclooxygenase-2
- DAPI:
-
4′, 6′-diamidine-2′-phenylindole dihydrochloride
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- ELISA:
-
Enzyme-linked immunosorbent assay
- ERK-1/-2:
-
Extracellular signal-regulated kinases-1/-2
- FITC:
-
Fluorescein iso-thiocyanate
- GAPDH:
-
Glyceraldehyde 3-phosphate dehydrogenase
- MAP kinase:
-
Mitogen-activated protein kinases
- NO:
-
Nitric oxide
- OA:
-
Osteoarthritis
- PBS:
-
Phosphate buffered saline
- RA:
-
Rheumatoid arthritis
- SNP:
-
Sodium nitroprusside
- SSZ:
-
Sulfasalazine.
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Lee, W.K., Kang, J.S. Modulation of Apoptosis and Differentiation by the Treatment of Sulfasalazine in Rabbit Articular Chondrocytes. Toxicol Res. 32, 115–121 (2016). https://doi.org/10.5487/TR.2016.32.2.115
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DOI: https://doi.org/10.5487/TR.2016.32.2.115