Toxicity of smoke extracts towards A549 lung cells: Role of acrolein and suppression by carbonyl scavengers
Toxicity of smoke extracts towards A549 lung cells: Role of acrolein and suppression by carbonyl scavengers
复制标题
DOI：
10.1016/j.cbi.2009.12.006
复制
发表时间：
2010-02-12
影响因子：
5.1
通讯作者：
Thompson, Colin A.
中科院分区：
文献类型：
--
作者：
Burcham, Philip C.;Raso, Albert;Thompson, Colin A.
有毒的三碳亲电试剂丙烯醛是在多种有机物（包括聚乙烯等合成聚合物）燃烧时形成的。虽然已知在烟雾吸入性损伤 (SII) 中发挥关键作用，但高剂量含丙烯醛烟雾的肺毒性的分子基础尚不清楚。因此，SII 的药物干预很难针对致病性烟雾毒物（例如丙烯醛）。本研究的首要目的是通过监测已知丙烯醛靶标的内收和丙烯醛诱导基因的表达来确认丙烯醛在烟雾提取物对 A549 肺细胞的急性毒性中的作用。第二个目的是评估羰基清除剂保护细胞靶标和阻止烟雾提取物毒性的能力。提取物是通过缓冲盐水捕集器通过闷烧聚乙烯释放的冒泡烟雾来制备的。用 2,4-二硝基苯肼衍生化后，通过 HPLC 估算提取物中的丙烯醛水平。提取物对 A549 细胞具有剧毒，与同等浓度的单独丙烯醛相比，会引起更大的 ATP 消耗。毒性伴随着几个细胞骨架靶标的明显羰基化。即波形蛋白和角蛋白-7、-8 和-18。蛋白质印迹显示，聚乙烯燃烧产物还上调了几种丙烯醛响应蛋白标记物，包括 GADD45 beta、NQO1、HMOX、Hsp70、Nur77 和 Egr1。几种羰基清除剂（亚硫酸氢盐、D-青霉胺、肼苯哒嗪和 1-肼基异喹啉）可强烈减弱烟雾提取物的毒性，其中亚硫酸氢盐可抑制中间丝目标的加合和交联。当使用实时监测细胞阻抗检测时，亚硫酸氢盐还抑制了烟雾提取物的细胞毒性。这些发现证实了丙烯醛在烟雾细胞毒性中的关键作用，并表明阻断丙烯醛毒性的药物值得进一步研究，作为针对 SII 的可能干预措施。 (C) 2009 Elsevier Ireland Ltd. 保留所有权利。
The noxious 3-carbon electrophile acrolein forms on combustion of diverse organic matter including synthetic polymers such as polyethylene. While known to play a key role in smoke inhalation injury (SII), the molecular basis for the pulmonary toxicity of high dose acrolein-containing smoke is unclear. As a result, drug interventions in SII are poorly directed against pathogenetic smoke toxicants such as acrolein. The first aim of this study was to confirm a role for acrolein in the acute toxicity of smoke extracts towards A549 lung cells by monitoring adduction of known acrolein targets and the expression of acrolein-inducible genes. A second aim was to evaluate carbonyl scavengers for their abilities to protect cell targets and block smoke extract toxicity. Extracts were prepared by bubbling smoke released by smouldering polyethylene through a buffered saline-trap. Acrolein levels in the extracts were estimated via HPLC after derivatisation with 2,4-dinitrophenylhydrazine. Extracts were highly toxic towards A549 cells, eliciting greater ATP depletion than an equivalent concentration of acrolein alone. The toxicity was accompanied by pronounced carbonylation of several cytoskeletal targets. namely vimentin and keratins-7, -8 and -18. Western blotting revealed that polyethylene combustion products also upregulated several acrolein-responsive protein markers, including GADD45 beta, NQO1, HMOX, Hsp70, Nur77 and Egr1. Several carbonyl scavengers (bisulfite, D-penicillamine. hydralazine and 1-hydrazinoisoquinoline) strongly attenuated smoke extract toxicity, with bisulfite suppressing both the adduction and cross-linking of intermediate filament targets. Bisulfite also suppressed the cytotoxicity of smoke extracts when detected using real-time monitoring of cellular impedance. These findings confirm a key role for acrolein in smoke cytotoxicity and suggest drugs that block acrolein toxicity deserve further investigation as possible interventions against SII. (C) 2009 Elsevier Ireland Ltd. All rights reserved.