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1.
The Journal of the Korean Rheumatism Association ; : 51-60, 2007.
Article in Korean | WPRIM | ID: wpr-78264

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.


Subject(s)
Humans , Caseins , Chondrocytes , DNA , Electrophoretic Mobility Shift Assay , Enzyme-Linked Immunosorbent Assay , Gelatin , Matrix Metalloproteinases , NF-kappa B , Osteoarthritis , Serum Albumin , Tumor Necrosis Factor-alpha
2.
Immune Network ; : 170-178, 2001.
Article in Korean | WPRIM | ID: wpr-41075

ABSTRACT

BACKGROUND: Korean mistletoe (Viscum album) extract has been found to posses immunostimulatory activity. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract might activate mouse peritoneal macrophages to produce interleukin 1 beta (IL-1 beta). METHODS: Hemagglutination assay was carried out to examine whether M11C contained a lectin or not . To know the effect of M11C on the production of IL-1 beta, the macrophages were treated by the M11C, and then collected the supernatant (M11C stimulated macrophages-conditioned media; MMCM). MMCM was analyzed for the IL-1 beta quantification and mRNA expression by means of ELISA and RT-PCR, respectively. RESULTS: Maximum effective dose and time of M11C on IL-1 beta production from macrophages were 20 micro gram/ml and 8 hours, respectively . This ELISA data was reconfirmed by immunoblotting assay . indicating that M11C is a good candidate for an immunomodulator. The dose and time dependent effect s of M11C on the expression of IL-1 beta mRNA from macrophages was also shown in expression of mRNA detected by RT-PCR. Treatment dose and time for the maximum expression of IL-1 beta mRNA were 20 micro gram/ml and 4 hours, respectively . Maximum gene expression of IL-1 beta was much earlier than maximum production of it. CONCLUSION: As results, Korean mistletoe extract, M11C, may be used for an immunomodulator. This will be able to make up for and solve the problems caused by existent immunoagent with many adverse effects through many other studies in future including one molecule extraction.


Subject(s)
Animals , Mice , Enzyme-Linked Immunosorbent Assay , Gene Expression , Hemagglutination , Immunoblotting , Interleukin-1beta , Macrophages , Macrophages, Peritoneal , Mistletoe , RNA, Messenger
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