Randall Matthew J, Haenen Guido R M M, Bouwman Freek G, van der Vliet Albert, Bast Aalt
Department of Toxicology, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands; Department of Pathology, College of Medicine, University of Vermont, 89 Beaumont Avenue, Burlington, VT 05405, USA.
Department of Toxicology, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands.
Toxicol Lett. 2016 Jan 5;240(1):43-9. doi: 10.1016/j.toxlet.2015.10.009. Epub 2015 Oct 19.
Chronic obstructive pulmonary disease (COPD) is the leading cause of cigarette smoke-related death worldwide. Acrolein, a crucial reactive electrophile found in cigarette smoke mimics many of the toxic effects of cigarette smoke-exposure in the lung. In macrophages, cigarette smoke is known to hinder histone deacetylases (HDACs), glucocorticoid-regulated enzymes that play an important role in the pathogenesis of glucocorticoid resistant inflammation, a common feature of COPD. Thus, we hypothesize that acrolein plays a role in COPD-associated glucocorticoid resistance. To examine the role of acrolein on glucocorticoid resistance, U937 monocytes, differentiated with PMA to macrophage-like cells were treated with acrolein for 0.5h followed by stimulation with hydrocortisone for 8h, or treated simultaneously with LPS and hydrocortisone for 8h without acrolein. GSH and nuclear HDAC activity were measured, or gene expression was analyzed by qPCR. Acrolein-mediated TNFα gene expression was not suppressed by hydrocortisone whereas LPS-induced TNFα expression was suppressed. Acrolein also significantly inhibited nuclear HDAC activity in macrophage-like cells. Incubation of recombinant HDAC2 with acrolein led to the formation of an HDAC2-acrolein adduct identified by mass spectrometry. Therefore, these results suggest that acrolein-induced inflammatory gene expression is resistant to suppression by the endogenous glucocorticoid, hydrocortisone.
慢性阻塞性肺疾病(COPD)是全球与香烟烟雾相关死亡的主要原因。丙烯醛是香烟烟雾中一种关键的活性亲电试剂,它模拟了肺部暴露于香烟烟雾中的许多毒性作用。在巨噬细胞中,已知香烟烟雾会阻碍组蛋白脱乙酰酶(HDACs),这是一种受糖皮质激素调节的酶,在糖皮质激素抵抗性炎症的发病机制中起重要作用,而糖皮质激素抵抗性炎症是COPD的一个常见特征。因此,我们假设丙烯醛在COPD相关的糖皮质激素抵抗中起作用。为了研究丙烯醛对糖皮质激素抵抗的作用,用佛波酯(PMA)将U937单核细胞分化为巨噬细胞样细胞,用丙烯醛处理0.5小时,然后用氢化可的松刺激8小时,或者在没有丙烯醛的情况下同时用脂多糖(LPS)和氢化可的松处理8小时。测量谷胱甘肽(GSH)和细胞核HDAC活性,或者通过定量聚合酶链反应(qPCR)分析基因表达。氢化可的松不能抑制丙烯醛介导的肿瘤坏死因子α(TNFα)基因表达,而LPS诱导的TNFα表达受到抑制。丙烯醛还显著抑制巨噬细胞样细胞中的细胞核HDAC活性。用丙烯醛孵育重组HDAC2导致形成一种通过质谱鉴定的HDAC2-丙烯醛加合物。因此,这些结果表明,丙烯醛诱导的炎症基因表达对内源性糖皮质激素氢化可的松的抑制具有抗性。