臺大醫院-內科部;臺大醫學院;Lai, N-S.N-S.LaiYu, C-L.C-L.YuYin, W-Y.W-Y.YinYu, H-C.H-C.YuHuang, H-B.H-B.HuangTung, C-H.C-H.TungLu, M-C.M-C.Lu余家利2014-02-142018-07-112014-02-142018-07-112012http://ntur.lib.ntu.edu.tw//handle/246246/258762Abnormal Ca2+-mediated signalling contributes to the pathogenesis of rheumatoid arthritis (RA). However, the potential implication of calcium channel blocker in RA remained unknown. We hypothesized that nifedipine, an L-type calcium channel blocker, combined with a calcineurin inhibitor, could suppress T cell activation via targeting different level of the Ca2+ signalling pathway. The percentage of activated T cells and the apoptotic rate of mononuclear cells (MNCs) was measured by flow cytometry. The MNC viability, cytokine production, cytosolic Ca2+ level and activity of the nuclear factor of activated T cells (NFAT) were measured by enzyme-linked immunosorbent assay (ELISA). The NFAT-regulated gene expression, including interleukin (IL)-2, interferon (IFN)-gamma and granulocytemacrophage colony-stimulating factor (GM-CSF), was measured by real-time polymerase chain reaction (PCR). We found that the percentage of activated T cells in anti-CD3 + anti-CD28-activated MNC was higher in RA patients. High doses of nifedipine (50 mu M) increased MNCs apoptosis, inhibited T cell activation and decreased T helper type 2 (Th1) (IFN-gamma)/Th2 (IL-10) cytokine production in both groups. The Ca2+ influx was lower in anti-CD3 + anti-CD28-activated MNC from RA patients than healthy volunteers and suppressed by nifedipine. When combined with a subtherapeutic dose (50 ng/ml) of cyclosporin, 1 mu M nifedipine suppressed the percentage of activated T cells in both groups. Moreover, this combination suppressed more IFN-gamma secretion and NFAT-regulated gene (GM-CSF and IFN-gamma) expression in RA-MNCs than normal MNCs via decreasing the activity of NFATc1. In conclusion, we found that L-type Ca2+ channel blockers and subtherapeutic doses of cyclosporin act additively to suppress the Ca2+-calcineurin-NFAT signalling pathway, leading to inhibition of T cell activity. We propose that this combination may become a potential treatment of RA.114 bytestext/htmlcalcineurincalcium channel blockercyclosporinnuclear factor of activated T cellsrheumatoid arthritishttp://ntur.lib.ntu.edu.tw/bitstream/246246/258762/1/index.html