| The study found that the pathogenesis of rheumatoid arthritis involved the cartilage extracellular matrix degradation process by a series of proteolytic enzyme including matrix metalloproteinases (MMPs), which is a large family of enzymes of degrading the various protein components in ECM especially MMP-1and MMP-9. In oriental medicines, melittin therapy is one of the more effective treatments of rheumatoid arthritis. Venom is mixture consisting of the various peptides, enzymes and other non-peptide, which melittin is the main active component of bee venom. The treatment process of melittin on rheumatoid arthritis may have connection with the expression of MMPs. Therefore, we studied the effects of melittin on the expression of MMPs by detecting the role of lipopolysaccharide (LPS)or tumor necrosis factor (TNF-α)in Raw264.7cells.[Objective] We study the effect of melittin on MMPs expression in Raw264.7cells.[Method] First, MTT assay detected the effect of different concentrations of LPS and TNF-α on cell viability by using subcultured Raw264.7cells. The highest concentrations of LPS and TNF-α, which didn’t decrease the cell viability, were selected as the experiment in the application of concentration. Then, ELISA was employed to detect MMPs extracellular secretion levels before and after LPS or TNF-αstimulating Raw264.7cells as well as the presence of different concentrations of melittin. RT-PCR was employed to detect MMPs intracellular expression levels in the same circumstances.[Results] Extracellular secretion levels and intracellular expression levels of MMPs have increased significantly by comparing with control group, any treatment Raw264.7cells as control group, after LPS or TNF-αstimulating cells. However, extracellular and intracellular expression levels were inhibited by melittin.[Conclusion] Melittin reduced the degradation of ECM contributing to protect cartilage and treat rheumatoid arthritis by inhibiting raw264.7cells and the inflammatory action of LPS or TNF-α. |
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