The Effects of Dehydroepiandrosterone-pyruvate on Human Osteoarthritic Chondrocytes.
- Author:
Myung Chul LEE
1
;
Jong Hun JI
;
Weon Yoo KIM
;
Jeong Hun DO
;
Yong Min KIM
Author Information
1. Department of Orthopedic Surgery, Seoul National University Hospital, Seoul, Korea.
- Publication Type:In Vitro ; Original Article
- Keywords:
Dehydroepiandrosterone-pyruvate;
Human Osteoarthritic Chondrocytes;
MMPs;
TIMP-1
- MeSH:
Cartilage;
Cartilage, Articular;
Chondrocytes*;
Dehydroepiandrosterone;
DNA;
Enzyme-Linked Immunosorbent Assay;
Gene Expression;
Humans*;
Interleukin-1;
Knee;
Matrix Metalloproteinases;
Osteoarthritis;
Tissue Inhibitor of Metalloproteinase-1
- From:Journal of Korean Orthopaedic Research Society
2006;9(2):153-164
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
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 micrometer 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 micrometer. 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 micrometer and 50 micrometer, 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.
