Division of Cardiology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, 114, Taiwan, R.O.C.
Institute of Cellular and System Medicine, National Health Research Institute, Zhunan 350, Taiwan, R.O.C.
Exp Biol Med (Maywood). 2015 Jan;240(1):99-108. doi: 10.1177/1535370214544263. Epub 2014 Jul 29.
Amiodarone, a common and effective antiarrhythmic drug, has been reported to have anti-inflammatory effects such as reducing the activation and movement of neutrophils. However, its effects on human T cells remain unclear. The aim of this study was to elucidate the effects and possible underlying mechanisms of amiodarone on human T cells. We isolated human primary T cells from the peripheral blood of healthy volunteers and performed enzyme-linked immunosorbent assay (ELISA), flow cytometry, electrophoretic mobility shift assay, luciferase assay, and Western blotting to evaluate the modulatory effects of amiodarone on human T cells. We found that amiodarone dose dependently inhibited the production of cytokines, including interleukin-2 (IL-2), IL-4, tumor necrosis factor-alpha, and interferon-gamma in activated human T cells. By flow cytometry, we demonstrated that amiodarone suppressed the expression of IL-2 receptor-alpha (CD25) and CD69, the cell surface markers of activated T cells. Moreover, molecular investigations revealed that amiodarone down-regulated activator protein-1 (AP-1) and nuclear factor kappa-B (NF-κB) DNA-binding activities in activated human T cells and also inhibited DNA binding and transcriptional activities of both AP-1 and NF-κB in Jurkat cells. Finally, by Western blotting, we showed that amiodarone reduced the activation of c-Jun NH(2)-terminal protein kinase and P38 mitogen-activated protein kinase, and suppressed stimuli-induced I-kappa B-alpha degradation in activated human T cells. Through regulation of AP-1 and NF-κB signaling, amiodarone inhibits cytokine production and T cell activation. These results show the pleiotropic effects of amiodarone on human T cells and suggest its therapeutic potential in inflammation-related cardiovascular disorders.
胺碘酮是一种常用且有效的抗心律失常药物,已被报道具有抗炎作用,如减少中性粒细胞的激活和运动。然而,其对人 T 细胞的影响尚不清楚。本研究旨在阐明胺碘酮对人 T 细胞的作用及其可能的机制。我们从健康志愿者的外周血中分离人原代 T 细胞,通过酶联免疫吸附试验(ELISA)、流式细胞术、电泳迁移率变动分析、荧光素酶分析和 Western blot 分析评估胺碘酮对人 T 细胞的调节作用。我们发现,胺碘酮呈剂量依赖性地抑制细胞因子的产生,包括激活的人 T 细胞中白细胞介素-2(IL-2)、IL-4、肿瘤坏死因子-α和干扰素-γ。通过流式细胞术,我们证实胺碘酮抑制了 IL-2 受体-α(CD25)和 CD69 的表达,这是激活的 T 细胞的表面标志物。此外,分子研究表明,胺碘酮下调激活的人 T 细胞中激活蛋白-1(AP-1)和核因子 kappa-B(NF-κB)DNA 结合活性,并抑制 Jurkat 细胞中 AP-1 和 NF-κB 的 DNA 结合和转录活性。最后,通过 Western blot,我们发现胺碘酮减少了 c-Jun NH(2)-末端蛋白激酶和 P38 丝裂原活化蛋白激酶的激活,并抑制了激活的人 T 细胞中刺激诱导的 I-kappa B-α降解。通过调节 AP-1 和 NF-κB 信号通路,胺碘酮抑制细胞因子的产生和 T 细胞的激活。这些结果显示了胺碘酮对人 T 细胞的多效性作用,并提示其在炎症相关心血管疾病中的治疗潜力。