J Korean Rheum Assoc.  2007 Mar;14(1):51-60. 10.4078/jkra.2007.14.1.51.

Advanced Glycation End Products Increase Matrix Metalloproteinases in Human Osteoarthritic Chondrocytes

• Nah SS
• Choi IY
• Mun SH
• Kim YG
• Moon HB
• Yoo B
• Lee CK

• KMID: 2202190
• DOI: http://doi.org/10.4078/jkra.2007.14.1.51

Abstract

OBJECTIVE
Although increased expression of receptor for advanced glycation end products (AGE) in osteoarthritis (OA) has been reported, little is known concerning the role of AGEs in the pathogenesis of OA. This study was undertaken to determine the effect of AGEs on the regulation of matrix metalloproteinase (MMP) expressions and activities in human OA chondrocytes
METHODS
OA chondrocytes were treated with increasing doses of AGE-bovine serum albumin (AGE-BSA). The expressions of MMPs were determined by both enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. The activities of MMPs were evaluated by both gelatin and casein zymography assays. In addition, electrophoretic mobility shift assay (EMSA) was employed to investigate the DNA binding activity of nuclear factor-kappa B (NF-kappaB) by AGE-BSA treatment.
RESULTS
The productions of MMP-1, -3, and -13 were significantly elevated by AGE-BSA in a dose dependent manner. The elevated activities of MMP-1, -3, and -13, and TNF-alpha by AGE-BSA were also observed. DNA binding activity of NF-kappaB was markedly increased by AGE-BSA treatment implicating possible involvement of NF-kappaB mediated pathway in the AGE-BSA induced MMP-1, -3, and -13, and TNF-alpha productions in OA chondrocytes. Taken together, this study demonstrates the stimulatory effect of AGE-BSA on the productions of MMPs and TNF-alpha and suggests the possible involvement of NF-kappaB mediated pathway in OA chondrocytes.
CONCLUSION
These results suggest that AGE may play a role in pathogenesis of OA.

Keyword

Advanced glycation end products; Matrix metalloproteinase; Chondrocyte; Osteoarthritis

MeSH Terms

Caseins
Chondrocytes*
DNA
Electrophoretic Mobility Shift Assay
Enzyme-Linked Immunosorbent Assay
Gelatin
Glycosylation End Products, Advanced*
Humans*
Matrix Metalloproteinases*
NF-kappa B
Osteoarthritis
Serum Albumin
Tumor Necrosis Factor-alpha
Caseins
DNA
Gelatin
Glycosylation End Products, Advanced
Matrix Metalloproteinases
NF-kappa B
Serum Albumin
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1. Immunocytochemical staining for type I and II collagen in monolayer-cultured osteoarthritis (OA) chondrocytes. Spherical or polygonal cells containing a single eccentric nucleus were observed. Type II (right panel), but not type I (left panel), collagen was detected in OA chondrocytes 7 days after primary culture (A). The effect of advanced glycation end product bovine serum albumin (AGE-BSA) on the viability of OA chondrocytes (B).

• 

  Fig. 2. Induction of nuclear factor-kappa B (NF- kB) (A), and tumor necrosis factor- α (TNF- α) expression by advanced glycation end product bovine serum albumin (AGE-BSA) in human osteoarthritis (OA) chondrocytes (B). n=4, ∗p<0.05 compaired with none and AGE-BSA or BSA treatment.

• 

  Fig. 3. Functional assay with zymography on matrix me- talloproteinase (MMPs) activity by advanced gly- cation end products (AGE) in human osteoarthritis (OA) chondrocytes. As MMP-1, 3, and 13 could be expressed in casein gels and have similar size, they were represented like one band.

• 

  Fig. 4. Induction of expression of matrix metalloproteinases (MMPs)-1, -3, and-13, by advanced glycation end product bovine serum albumin (AGE-BSA). The expression levels of MMP-1, -3, and-13 were also analyzed by Western-blot assays (A). Enzyme linked immunosorbent assay (ELISA) performed for productions of MMP-1 (upper panel), MMP-3 (middle panel), and MMP-13 (lower panel) (B). Results are representative of duplicate experiments. n=6, ∗p<0.05. compaired with none and AGE-BSA or BSA treatment. p<0.05. compaired with same dose of BSA.

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