Department of Pharmacotherapeutics, Graduate School of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 729-0251, Japan; Departments of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 729-0251, Japan.
Departments of Pharmacotherapeutics, Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 985-1 Sanzo, Higashimura-cho, Fukuyama, Hiroshima 729-0251, Japan.
Eur J Pharmacol. 2019 Jan 5;842:33-39. doi: 10.1016/j.ejphar.2018.10.029. Epub 2018 Oct 23.
We investigated the effects of shogaol, which has an α, β-unsaturated carbonyl group, and gingerol, which does not, on primary-cultured microglia to understand how the α, β-unsaturated carbonyl interacts with Kelch-like ECH-associated protein (Keap)1. Shogaol (1 μM) but not the same concentration of gingerol significantly increased heme oxygenase (HO)-1 protein levels in cultured microglia without cytotoxicity. In addition, shogaol suppressed the release of the inflammation marker nitric oxide induced by 30 U/ml thrombin treatment. A docking simulation suggested that the α, β-unsaturated carbonyl of shogaol but not gingerol interacts with Keap1. Nuclear import of nuclear factor E2-related factor 2 and increased binding of the HO-1 E2 enhancer support the docking-simulation prediction. The transcription inhibitor actinomycin D (0.1 μg/ml) markedly blocked the increase of HO-1 mRNA levels by shogaol. To evaluate whether the α, β-unsaturated carbonyl can be used for intracerebral hemorrhage (ICH) therapy, we investigated the effect of shogaol on an in vivo mouse ICH model. Intracerebroventricular injection of 0.2 nmol shogaol increased striatal HO-1 protein levels and rescued ICH-induced neuron loss. Thus, the α, β-unsaturated carbonyl is necessary for the interaction of compounds, such as shogaol, with Keap1, and these findings may be useful for screening novel ICH therapeutic agents that increase HO-1 expression.
我们研究了具有α,β-不饱和羰基的姜烯酚和没有α,β-不饱和羰基的姜酚对原代培养的小胶质细胞的影响,以了解α,β-不饱和羰基如何与Kelch 样 ECH 相关蛋白 1(Keap1)相互作用。姜烯酚(1 μM)而非相同浓度的姜酚可在无细胞毒性的情况下显著增加培养小胶质细胞中的血红素加氧酶(HO)-1 蛋白水平。此外,姜烯酚抑制了 30 U/ml 凝血酶处理诱导的炎症标志物一氧化氮的释放。对接模拟表明,姜烯酚的α,β-不饱和羰基而不是姜酚与 Keap1 相互作用。核因子 E2 相关因子 2 的核导入和 HO-1 E2 增强子结合的增加支持对接模拟预测。转录抑制剂放线菌素 D(0.1μg/ml)显著阻断了姜烯酚对 HO-1 mRNA 水平的增加。为了评估α,β-不饱和羰基是否可用于脑出血(ICH)治疗,我们研究了姜烯酚对体内小鼠 ICH 模型的影响。脑室注射 0.2 nmol 姜烯酚可增加纹状体 HO-1 蛋白水平并挽救 ICH 诱导的神经元丢失。因此,α,β-不饱和羰基是化合物(如姜烯酚)与 Keap1 相互作用所必需的,这些发现可能有助于筛选增加 HO-1 表达的新型 ICH 治疗剂。