Research Center of Ion Channelopathy, Institute of Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R. China.
PLoS One. 2013 May 22;8(5):e64629. doi: 10.1371/journal.pone.0064629. Print 2013.
Diphenyl phosphine oxide-1 (DPO-1) is a potent Kv1.5 channel inhibitor that has therapeutic potential for the treatment of atrial fibrillation. Many other Kv1.5 channel blockers also potently inhibit the Kv1.3 channel, but whether DPO-1 blocks Kv1.3 channels has not been investigated. The Kv1.3 channel is highly expressed in activated T cells, which is considered a favorable target for immunomodulation. Accordingly, we hypothesized that DPO-1 may exert immunosuppressive and anti-inflammatory effects by inhibiting Kv1.3 channel activity. In this study, DPO-1 blocked Kv1.3 current in a voltage-dependent and concentration-dependent manner, with IC₅₀ values of 2.58 µM in Jurkat cells and 3.11 µM in human peripheral blood T cells. DPO-1 also accelerated the inactivation rate and negatively shifted steady-state inactivation. Moreover, DPO-1 at 3 µM had no apparent effect on the Ca²⁺ activated potassium channel (K(Ca)) current in both Jurkat cells and human peripheral blood T cells. In Jurkat cells, pre-treatment with DPO-1 for 24 h decreased Kv1.3 current density, and protein expression by 48±6% and 60±9%, at 3 and 10 µM, respectively (both p<0.05). In addition, Ca²⁺ influx to Ca²⁺-depleted cells was blunted and IL-2 production was also reduced in activated Jurkat cells. IL-2 secretion was also inhibited by the Kv1.3 inhibitors margatoxin and charybdotoxin. Our results demonstrate for the first time that that DPO-1, at clinically relevant concentrations, blocks Kv1.3 channels, decreases Kv1.3 channel expression and suppresses IL-2 secretion. Therefore, DPO-1 may be a useful treatment strategy for immunologic disorders.
二苯基膦氧化物-1(DPO-1)是一种有效的 Kv1.5 通道抑制剂,具有治疗心房颤动的潜力。许多其他 Kv1.5 通道阻滞剂也能强烈抑制 Kv1.3 通道,但 DPO-1 是否阻断 Kv1.3 通道尚未得到研究。Kv1.3 通道在激活的 T 细胞中高度表达,被认为是免疫调节的有利靶点。因此,我们假设 DPO-1 通过抑制 Kv1.3 通道活性可能发挥免疫抑制和抗炎作用。在这项研究中,DPO-1 以电压依赖性和浓度依赖性方式阻断 Kv1.3 电流,在 Jurkat 细胞中的 IC₅₀值为 2.58 µM,在人外周血 T 细胞中的 IC₅₀值为 3.11 µM。DPO-1 还加速了失活速率并负移了稳态失活。此外,在 Jurkat 细胞和人外周血 T 细胞中,3 µM 的 DPO-1 对 Ca²⁺激活钾通道(K(Ca))电流没有明显影响。在 Jurkat 细胞中,用 DPO-1 预处理 24 小时后,3 和 10 µM 时 Kv1.3 电流密度和蛋白表达分别降低 48±6%和 60±9%(均 p<0.05)。此外,Ca²⁺内流到 Ca²⁺耗尽的细胞被削弱,激活的 Jurkat 细胞中 IL-2 的产生也减少。Kv1.3 抑制剂 margatoxin 和 charybdotoxin 也抑制了 IL-2 的分泌。我们的研究结果首次表明,在临床相关浓度下,DPO-1 阻断 Kv1.3 通道,降低 Kv1.3 通道表达并抑制 IL-2 分泌。因此,DPO-1 可能是治疗免疫紊乱的一种有用策略。