Objectives : This study is aimed to investigate the effects of bee venom and melittin on cell death in synovial cell line.
Methods : It was evaluated by using MTT assay, morphologic method, DNA fragmenation, NO generation, flow cytometry, immunocytochemistry analysis, RT-PCR, Western blot.
Results : The obtained results are summarized as follows:
1. The MTT assay demonstrated that synovial cell viability was significantly inhibitted dose-dependently by treatment with bee venom and melittin in comparison with control.
2. The morphologic study demonstrated that synovial cell showed apoptosis after treatment with bee venom and melittin for 6 hours using microscope.
3. In case of NO generation bee venom group and melittin group showed significant inhibition in comparison with control.
4. The Flow cytometry demonstrated that apoptosis of synovial cell treated with bee venom and melittin was related with stop of cell cycle in stage of G_0/G_1.
5. DNA fragmenation demonstrated that synovial cell treated with bee venom and melittin showed DNA ladder below l Kbp.
6. Immunocytochemistry assay demonstrated that COX-Ⅱ and PLA2 were strongly down-regulated by treatment with bee venom and melittin whereas iNOS was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment.
7. RT-PCR analysis demonstrated that iNOS were strongly down-regulated by treatment with bee venom and melittin whereas COX-Ⅱ was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment.
8. Western blot demonstrated that iNOS were strongly down-regulated by treatment with 15 ㎍/ml bee venom whereas COX-Ⅱ was strongly down-regulated from 5 ㎍/ml bee venom.
Conclusions : These results suggest that bee venom and melittin have significant effect on cell death in synovial cell line and further study is needed in vivo.
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