Ambient particulate matter affects occludin distribution and increases alveolar transepithelial electrical conductance.
Ambient particulate matter affects occludin distribution and increases alveolar transepithelial electrical conductance.
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DOI：
10.1111/j.1440-1843.2010.01910.x
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发表时间：
2011-02
期刊：
影响因子：
--
通讯作者：
Comellas AP
中科院分区：
文献类型：
--
作者：
Caraballo JC;Yshii C;Westphal W;Moninger T;Comellas AP
本研究在体外研究了颗粒物质和柴油机尾气颗粒对肺泡屏障完整性的影响。我们的研究结果表明，这些颗粒改变了紧密连接的完整性，特别是Occludin和ZO-1的结合。这些影响可以通过阻断线粒体ROS的产生来阻止，这表明该细胞器在观察到的影响中起着核心作用。吸入颗粒物（PM）引起肺部炎症和上皮功能障碍。然而，PM对肺泡上皮屏障完整性的直接影响尚不清楚。我们的目的是确定PM暴露是否会影响肺泡上皮细胞（AEC）、经上皮电导（Gt）和紧密连接（TJ）蛋白。将人类AEC （A549）和原代大鼠AEC暴露于粒径<10 μm的PM （PM10）和柴油尾气颗粒（DEP）中，并以二氧化钛（TiO2）作为粒径效应的对照。测定Gt和对异硫氰酸酯-葡聚糖荧光素（fitc -葡聚糖）的通透性以评估屏障完整性。透射电镜下通过分析硝酸镧（La3+）的穿透性来评价TJ的完整性。表面蛋白用生物素标记，Western blot （WB）分析。免疫荧光法评估TJ蛋白包括occludin和zonula occludens-1 （ZO-1）的共定位。用共免疫沉淀法评价PM诱导occludin-ZO-1的解离。治疗3h后，PM10和DEP增加Gt，破坏TJ。PM10和DEP诱导occludin从质膜内化到内体腔室和occludin从ZO-1分离。抗氧化酶锰超氧化物歧化酶（MnSOD）和过氧化氢酶的过度表达，阻止了pm诱导的Gt增加、质膜occludin减少及其与ZO-1的分离。PM部分通过质膜occludin减少和AEC中ZO-1解离诱导肺泡上皮功能障碍。此外，这些作用可通过两种不同的抗氧化酶的过表达来阻止。
This work studies the effects of particulate matter and diesel exhaust particles on alveolar barrier integrity in vitro. Our results show that these particles alter tight junction integrity, specifically the Occludin and ZO-1 association. These effects are prevented by blocking of mitochondrial ROS production, suggesting a central role of this organelle in the effects observed. Inhaled particulate matter (PM) causes lung inflammation and epithelial dysfunction. However, the direct effect of PM on alveolar epithelial barrier integrity is not well understood. Our aim is to determine whether PM exposure affects the alveolar epithelial cells (AEC) transepithelial electrical conductance (Gt) and tight junction (TJ) proteins. Human AEC (A549) and primary rat AEC were exposed to PM of <10 μm in size (PM10) and diesel exhaust particles (DEP), using titanium dioxide (TiO2) as a control for particle size effects. Gt and permeability to fluorescein isothiocyanate-dextran (FITC-dextran) were measured to assess barrier integrity. TJ integrity was evaluated by analyzing penetration of Lanthanum nitrate (La3+) under transmission electron microscopy. Surface proteins were labeled with biotin and analyzed by Western blot (WB). Immunofluorescence was performed to assess co-localization of TJ proteins including occludin and zonula occludens-1 (ZO-1). PM induced dissociation of occludin-ZO-1 was evaluated by co-immunoprecipitation. PM10 and DEP increased Gt and disrupted TJ after 3h of treatment. PM10 and DEP induced occludin internalization from the plasma membrane into endosomal compartments and dissociation of occludin from ZO-1. Overexpression of antioxidant enzymes Manganese Superoxide Dismutase (MnSOD) and Catalase, prevented PM-induced Gt increase, occludin reduction from the plasma membrane and its dissociation from ZO-1. PM induces alveolar epithelial dysfunction in part via occludin reduction at the plasma membrane and ZO-1 dissociation in AEC. Furthermore, these effects are prevented by overexpression of two different antioxidant enzymes.
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影响因子：
37.8
作者：
Pope, CA;Burnett, RT;Godleski, JJ
通讯作者：
Godleski, JJ
影响因子：
4.4
作者：
Phillips, Brett E.;Cancel, Limary;Antonetti, David A.
通讯作者：
Antonetti, David A.
影响因子：
10
作者：
Lehmann, Andrea D.;Blank, Fabian;Rothen-Rutishauser, Barbara M.
通讯作者：
Rothen-Rutishauser, Barbara M.
影响因子：
14.2
作者：
O'Connor, George T.;Neas, Lucas;Lippmann, Morton
通讯作者：
Lippmann, Morton
DOI：
10.1152/ajpcell.2000.279.1.c21
发表时间：
2000-07-01
影响因子：
5.5
作者：
Kevil, CG;Oshima, T;Alexander, JS
通讯作者：
Alexander, JS