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KMID : 0869020060090020153
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Journal of Korean Orthopaedic Research Society
2006 Volume.9 No. 2 p.153 ~ p.164
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The Effects of Dehydroepiandrosterone-pyruvate on Human Osteoarthritic Chondrocytes
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Lee Myung-Chul
Ji Jong-Hun
Kim Weon-Yoo
Do Jeong-Hun
Kim Yong-Min
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Abstract
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Purpose: To investigate the in vitro effects of Dehydroepiandrosterone (DHEA)-pyruvate on human osteoarthritic chondrocytes.
Methods: Chondrocytes isolated from human osteoarthritic knee cartilage were three-dimensionally cultured in Alginate beads. Cells were treated with dehydroepiandrosterone in the presence or absence of IL-1. The effects on chondrocytes were analyzed by MTS assay (for chondrocytes proliferation), DMB assay (for glycosaminoglycan synthesis), and indole assay (for DNA amount). Gene expressions of MMP-1,3, TIMP-1 as well as IL-1 induced gene expression of MMP-1, 3 were analyzed using RT-PCR. The protein synthesis of MMP-1,3 and TIMP-1 was determined by ELISA.
Results: Treatment of chondrocytes with DHEA-pyruvate did not affect chondrocytes proliferation regardless of concentrations when compared with control. GAG synthesis was not changed significantly during culture period regardless of concentrations. DHEA-pyruvate suppressed the expression of MMP-1 significantly at a concentration of 50 μM and above. The gene expression of MMP-3 was also suppressed. The expression of TIMP-1 was significantly increased by DHEA-pyruvate at concentration of 50 μM. The effects of DHEA-pyruvate were also prominent in the presence of IL-1, in which IL-1induced gene expressions of not only MMP-1, but also MMP-3 were suppressed at a lower concentration of 10 μM and 50 μM, respectively. In enzyme activity measurement, the results came in line with the results obtained by RT-PCR, which means DHEA-pyruvate influences on the gene transcription level.
Conclusion: Our study clearly demonstrated that DHEA-pyruvate has the ability to modulate the imbalance between MMPs and TIMP-1 during osteoarthritis at the transcription level, suggesting its protective role against loss of articular cartilage.
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KEYWORD
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Dehydroepiandrosterone-pyruvate, Human Osteoarthritic Chondrocytes, MMPs, TIMP-1
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