Comparison of wood smoke PM2.5 obtained from the combustion of FIR and beech pellets on inflammation and DNA damage in A549 and THP-1 human cell lines
Comparison of wood smoke PM2.5 obtained from the combustion of FIR and beech pellets on inflammation and DNA damage in A549 and THP-1 human cell lines
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DOI：
10.1007/s00204-013-1071-z
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发表时间：
2013-12-01
影响因子：
6.1
通讯作者：
Galli, Corrado L.
中科院分区：
文献类型：
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作者：
Corsini, Emanuela;Budello, Silvia;Galli, Corrado L.
本研究的目的是调查白细胞介素-8的诱导颗粒物（PM）从冷杉和山毛榉颗粒在家用电器上两个人类细胞系，即肺上皮细胞系A549和早幼粒细胞系THP-1燃烧的效果。从燃烧山毛榉和冷杉颗粒获得的PM2.5的效果进行了比较，以参考柴油机排气颗粒物（DEP）。同时，木烟雾PM诱导的遗传毒性和氧化应激也在A549细胞中进行了研究。细胞处理不同的时间（3-72小时）与浓度增加的PM2.5从顺序燃烧的冷杉和山毛榉颗粒或参考DEP。通过乳酸脱氢酶渗漏评估细胞活力，并测量白细胞介素-8或CXCL 8（IL-8）的释放以评估促炎作用。采用5（6）-羧基-2 '，7'-二氯荧光素二乙酸酯（DCFH-DA）试验评价氧化应激，采用碱性彗星试验评价DNA损伤，采用流式细胞术评价微核率。A549和THP-1细胞都以剂量和时间相关的方式对木材烟雾PM2.5产生IL-8释放，从后期燃烧中获得的颗粒最活跃。THP-1细胞比A549细胞更敏感。在质量基础上，观察到类似的效果，冷杉和山毛榉PM2.5。然而，燃烧山毛榉颗粒产生的PM2.5比冷杉颗粒多约三倍。关于PM2.5摄取的机制，在THP-1和A549细胞中，细胞松弛素D阻止PM2.5诱导的IL-8 mRNA表达和细胞因子释放，表明肌动蛋白聚合在颗粒摄取中起关键作用，并且IL-8的产生与颗粒吞噬相关。在THP-1和A549细胞中，选择性抑制剂SB 203580可完全阻断PM2.5诱导的IL-8释放，提示p38 MAPK的激活作用。从冷杉和山毛榉颗粒PM2.5也诱导适度的DNA损伤剂量相关的，测量为链断裂，而没有观察到微核数量的增加。DEP也观察到了类似的效果，反对木材烟雾颗粒的危险性低于相同尺寸范围内其他类别的燃烧产生的颗粒。总的来说，结果表明，燃烧条件可以显着影响颗粒的特性和随之而来的毒性，不同的木材可以产生不同数量的PM2.5。
The aim of this study was to investigate the effect on the induction of interleukin-8 of particulate matter (PM) from fir and beech pellets burnt in domestic appliances on two human cells lines, namely the lung epithelial cell line A549 and the promyelocytic cell line THP-1. The effects of PM2.5 obtained from combustion of beech and fir pellets were compared to reference diesel exhaust particulates (DEP). In parallel, wood smoke PM-induced genotoxicity and oxidative stress were also investigated in A549 cells. Cells were treated for different times (3-72 h) with increasing concentrations of PM2.5 obtained from sequential combustions of fir and beech pellets or reference DEP. Cell viability was assessed by lactate dehydrogenase leakage, and the release of interleukin-8 or CXCL8 (IL-8) was measured to evaluate the pro-inflammatory effect. Oxidative stress was evaluated by the 5(6)-carboxy-2',7'dichlorofluorescein diacetate (DCFH-DA) assay and DNA damage by the alkaline comet assay and micronucleus frequency by flow cytometry. Both A549 and THP-1 cells responded in a dose- and time-related manner to wood smoke PM2.5 with IL-8 release, particles obtained from late combustions being the most active. THP-1 cells were more sensitive than A549 cells. On a mass base, similar effects were observed for both fir and beech PM2.5. However, the combustion of beech pellets generated approximately three times more PM2.5 than fir pellets. Regarding the mechanism of PM2.5 uptake, in both THP-1 and A549 cells, cytochalasin D prevented PM2.5-induced IL-8 mRNA expression and cytokine release, indicating a key role for actin polymerization in particles uptake and that the production of IL-8 correlated with particle phagocytosis. As signal transduction pathway involvement, in both THP-1 and A549 cells, PM2.5-induced IL-8 release could be completely blocked by the selective inhibitor SB203580, indicating a role of p38 MAPK activation. PM2.5 from both fir and beech pellets also induced modest DNA lesions dose related, measured as strand breaks, whereas no increase in the number of micronucleus was observed. Similar effects were observed with DEP, arguing against less dangerous effects of wood smoke particles than other categories of combustion-derived particles in the same size range. Overall, results suggest that combustion conditions can significantly affect the characteristics of particles and the consequent toxicity, and that different woods can generate different amounts of PM2.5.