Proteases and oxidant stress control organic dust induction of inflammatory gene expression in lung epithelial cells.
Proteases and oxidant stress control organic dust induction of inflammatory gene expression in lung epithelial cells.
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DOI：
10.1186/s12931-016-0455-z
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发表时间：
2016-10-22
影响因子：
5.8
通讯作者：
Boggaram V
中科院分区：
文献类型：
--
作者：
Natarajan K;Gottipati KR;Berhane K;Samten B;Pendurthi U;Boggaram V
对有机粉尘中的感染因子和其他成分的持续炎症反应是肺损伤和呼吸道疾病发展的基础。引起炎症的有机粉尘成分及其引起肺部炎症的机制尚未完全了解。我们研究了家禽粉尘提取物中的蛋白酶活性和细胞内氧化应激诱导A549和Beas 2B肺上皮细胞中炎症基因表达的机制。通过定量聚合酶链反应（qPCR）、酶联免疫吸附（ELISA）和蛋白质印迹分析粉尘提取物对炎症基因表达的影响。通过二氢乙锭（DHE）标记和4-羟基壬烯醛（4-HNE）的免疫染色来探测氧化应激。通过瞬时转染试验测定对白细胞介素-8（IL-8）启动子调节的影响。粉尘提取物含有胰蛋白酶和弹性蛋白酶活性，并激活蛋白酶激活受体（PAR）-1和-2。丝氨酸蛋白酶抑制剂和PAR-1或PAR-2敲低抑制炎症基因诱导。粉尘提取物诱导IL-8基因表达与DHE荧光和4-HNE染色增加相关，抗氧化剂抑制炎症基因诱导。蛋白酶抑制剂和抗氧化剂抑制蛋白激酶C和NF-κB激活和IL-8启动子活性的诱导细胞暴露于粉尘提取物。我们的研究表明，蛋白酶和细胞内氧化剂控制有机粉尘诱导肺上皮细胞中的炎症基因表达。靶向蛋白酶和氧化应激可能成为治疗有机粉尘所致肺部疾病的新方法。这是第一份关于氧化应激参与有机粉尘诱导炎症基因表达的报告。本文的在线版本（doi：10.1186/s12931-016-0455-z）包含补充材料，可供授权用户使用。
Persistant inflammatory responses to infectious agents and other components in organic dust underlie lung injury and development of respiratory diseases. Organic dust components responsible for eliciting inflammation and the mechanisms by which they cause lung inflammation are not fully understood. We studied the mechanisms by which protease activities in poultry dust extracts and intracellular oxidant stress induce inflammatory gene expression in A549 and Beas2B lung epithelial cells. The effects of dust extracts on inflammatory gene expression were analyzed by quantitative polymerase chain reaction (qPCR), enzyme linked immunosorbent (ELISA) and western blot assays. Oxidant stress was probed by dihydroethidium (DHE) labeling, and immunostaining for 4-hydroxynonenal (4-HNE). Effects on interleukin-8 (IL-8) promoter regulation were determined by transient transfection assay. Dust extracts contained trypsin and elastase activities, and activated protease activated receptor (PAR)-1 and -2. Serine protease inhibitors and PAR-1 or PAR-2 knockdown suppressed inflammatory gene induction. Dust extract induction of IL-8 gene expression was associated with increased DHE-fluorescence and 4-HNE staining, and antioxidants suppressed inflammatory gene induction. Protease inhibitors and antioxidants suppressed protein kinase C and NF-κB activation and induction of IL-8 promoter activity in cells exposed to dust extract. Our studies demonstrate that proteases and intracellular oxidants control organic dust induction of inflammatory gene expression in lung epithelial cells. Targeting proteases and oxidant stress may serve as novel approaches for the treatment of organic dust induced lung diseases. This is the first report on the involvement of oxidant stress in the induction of inflammatory gene expression by organic dust. The online version of this article (doi:10.1186/s12931-016-0455-z) contains supplementary material, which is available to authorized users.
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影响因子：
6
作者：
Christian F;Smith EL;Carmody RJ
通讯作者：
Carmody RJ
影响因子：
3.6
作者：
Cederqvist, K;Haglund, C;Andersson, S
通讯作者：
Andersson, S
影响因子：
4.4
作者：
Asokananthan, N;Graham, PT;Stewart, GA
通讯作者：
Stewart, GA
影响因子：
3.1
作者：
Chuquimia, Olga D.;Petursdottir, Dagbjort H.;Fernandez, Carmen
通讯作者：
Fernandez, Carmen
DOI：
10.1164/ajrccm.150.4.7921472
发表时间：
1994-10-01
影响因子：
24.7
作者：
LARSSON, KA;EKLUND, AG;MALMBERG, PO
通讯作者：
MALMBERG, PO