Comparison of the Activities of NF-kappa B and I kappa B alpha in Patients with Rheumatoid Arthritis and Osteoarthritis / 대한정형외과연구학회지

PURPOSE: The nuclear factor-kappa B (NF-kappa B) has been known to regulate the inflammatory and immune process by transcription of inflammatory intermediates. The purpose of the present study is to show the difference in activity of NF-kappa B and its inhibitory factor-I kappa B alpha in patients with rheumatoid arthritis, osteoarthritis and normal control subjects. MATERIALS AND METHODS: Synovial membrane samples were obtained at the time of orthopedic surgery from the knees of 7 patients with RA and 7 patients with OA. Two control samples were obtained from an amputee with no history of arthritis. We designed the primer of the subunit p65 of NF-kappa B and I kappa B alpha, measured the activity of them by RT-PCR, and analyzed the expression of NF-kappa B by immunohistochemical staining. RESULTS: From the results of RT-PCR, the expression levels of NF-kappa B was found to be higher in synovial tissues obtained from patients with RA than from synovial tissue obtained from patients with OA, and the least from the control group. The expression levels of I kappa B alpha were not different statistically among the three groups. Immunohistochemical staining for the NF-kappa B was dominant in synovial tissue from patients with RA. The result of immunohistochemical staining was similar to the results of RT-PCR for NF-kappa B. The localization of the staining was predominantly nuclear. CONCLUSION: In this study, activity of NF-kappa B of rheumatoid arthritis was higher than the other group, but expressions of I kappa B alpha were no different between the diseases. Further studies about specific inhibitors of NF-kappa B will benefit the development of rheumatoid arthritis regimens with greater efficacy.

Effect of FK506 and Cyclosporin A on I(kappa)B(alpha) Degradation and IKK Pathway in Bronchial Epithelial Cells, Monocytes, Lymphocytes and Alveolar Macrophages / 결핵

BACKGROUND: Cyclosporin A(CsA) and tacrolimus(FK506) have been widely used as immunosuppressants. The effects of CsA, or FK506, on the IkappaB/NF-kappaB pathway have been shown to vary according to the cell type. However, their effects on the IkappaB/NF-kappaB pathway have not been reported in bronchial epithelial cells. In this study, the effects of CsA and FK506 on the IkappaB/NF-kappaB pathway in bronchial epithelial cells, monocytes, lymphocytes and alveolar macrophages were evaluated. The relationship between their effects on the IkappaB/NF-kappaB pathway and IkappaB kinase(IKK) activity was also investigated. METHODS: BEAS-2B and A549 cells, pulmonary alveolar macrophages, peripheral blood monocytes and lymphocytes were used. The cells were pre-treated with CsA, or FK506, for various time periods, followed by stimulation with TNF-alpha, LPS or IL-1beta. The I(kappa)B(alpha) expressions were assayed by Western blot analyses. The IKK activity was evaluated by an in vitro immune complex kinase assay, using GST-I(kappa)B(alpha) as the substrate. RESULTS: Neither CsA nor FK506 affected the level of I(kappa)B(alpha) expression in any of the cell types used in this study. CsA pre-treatment inhibited the TNFalpha-induced I(kappa)B(alpha) degradation in bronchial epithelial cells. In contrast, the TNFalpha-induced I(kappa)B(alpha) degradation was not affected by FK506 pre-treatment. However, FK506 suppressed the cytokine-induced I(kappa)B(alpha) degradation in the pulmonary alveolar macrophages, peripheral blood monocytes and lymphocytes. The inhibitory effect of CsA, or FK506, on I(kappa)B(alpha) degradation was not related to IKK. CONCLUSIONS: CsA and FK506 suppressed the I(kappa)B(alpha) degradation in bronchial epithelial cells, mono. cytes, lymphocytes and alveolar macrophages, so this may not be mediated through IKK.