Anti-inflammatory properties of cilostazol: Its interruption of DNA binding activity of NF-κB from the Toll-like receptor signaling pathways
Anti-inflammatory properties of cilostazol: Its interruption of DNA binding activity of NF-κB from the Toll-like receptor signaling pathways
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西洛他唑的抗炎特性：它通过 Toll 样受体途径信号阻断 NF-κB 的 DNA 结合活性
DOI：
10.1016/j.intimp.2018.06.021
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发表时间：
2018
影响因子：
5.6
通讯作者：
Hattori Y
中科院分区：
文献类型：
--
作者：
Sakamoto T;Ohashi W;Tomita K;Hattori K;Matsuda N;Hattori Y
西洛他唑是一种选择性磷酸二酯酶III抑制剂，具有抗血小板、抗促有丝分裂和血管舒张特性，广泛用于治疗外周血管疾病的缺血症状。大量证据表明，西洛他唑也表现出抗炎作用，但其抗炎机制尚未完全了解。在此，我们发现西洛他唑特异性抑制细胞因子的表达，这是由核因子-κB（NF-κB）激活诱导，在RAW 264.7巨噬细胞用不同的Toll样受体（TLR）配体刺激。发现西洛他唑显著降低TLR-4和TLR-3配体刺激的NF-κB转录活性，其通过荧光素酶报告基因测定进行定量。然而，西洛他唑对TLR-4配体脂多糖（LPS）刺激后IκBα降解和NF-κB p65磷酸化及核转位无影响。西洛他唑也不能阻止LPS诱导的丝裂原活化蛋白激酶（MAPK）家族磷酸化水平的增加。另一方面，使用染色质免疫沉淀试验，我们证明西洛他唑通过阻止NF-κB p65募集到这些基因启动子来降低LPS诱导的白细胞介素-6和肿瘤坏死因子-α的转录活性。当小鼠连续7天每天经口灌胃给予西洛他唑时，LPS诱导的异常促炎细胞因子产生和终末器官组织损伤显著减少。本研究的结果表明，西洛他唑能够直接中断TLR信号通路中NF-κB蛋白的DNA结合活性。特异性干预这一途径的治疗可能对预防不同的炎症性疾病有潜在的益处。
Cilostazol, a selective inhibitor of phosphodiesterase type III with anti-platelet, anti-mitogenic, and vasodilating properties, is widely used to treat ischemic symptoms of peripheral vascular disease. Ample evidence has suggested that cilostazol also exhibits an anti-inflammatory effect, but its anti-inflammatory mechanism is not fully understood. Here, we showed that cilostazol specifically inhibited expression of cytokines, which are induced by nuclear factor-κB (NF-κB) activation, in RAW264.7 macrophage cells stimulated with different Toll-like receptor (TLR) ligands. Cilostazol was found to significantly reduce TLR-4 and TLR-3 ligands-stimulated NF-κB transcriptional activity, which was quantified by luciferase reporter assays. However, cilostazol was without effect on IκBα degradation and NF-κB p65 phosphorylation and nuclear translocation after challenge with the TLR-4 ligand lipopolysaccharide (LPS). Cilostazol did not also prevent the LPS-induced increase in phosphorylated levels of the mitogen-activated protein kinase (MAPK) family. On the other hand, using chromatin immunoprecipitation assays, we demonstrated that cilostazol reduced the LPS-induced transcriptional activities of interleukin-6 and tumor necrosis factor-α by preventing the recruitment of NF-κB p65 to these gene promoters. When cilostazol was given to mice by oral gavage daily for 7 days, LPS-induced aberrant pro-inflammatory cytokine production and end-organ tissue injury were significantly reduced. The results of this study suggest that cilostazol is capable of directly interrupting DNA binding activity of NF-κB proteins from the TLR signaling pathways. The therapy to specifically intervene in this pathway may be potentially beneficial for the prevention of different inflammatory disorders.