Cellular and molecular mechanisms of cigarette smoke-induced lung damage and prevention by vitamin C.
Cellular and molecular mechanisms of cigarette smoke-induced lung damage and prevention by vitamin C.
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DOI：
10.1186/1476-9255-5-21
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发表时间：
2008-11-11
影响因子：
5.1
通讯作者：
Chatterjee, Indu B.
中科院分区：
文献类型：
--
作者：
Banerjee, Shuvojit;Chattopadhyay, Ranajoy;Ghosh, Arunava;Koley, Hemanta;Panda, Koustubh;Roy, Siddhartha;Chattopadhyay, Dhrubajyoti;Chatterjee, Indu B.
香烟烟雾引起肺损伤的细胞和分子机制尚不清楚。香烟烟雾是一种复杂的混合物，含有长寿命的自由基，包括导致氧化损伤的对苯醌。早先我们曾报道过氧化蛋白损伤是烟雾引起的肺损伤的初始事件。考虑到对苯醌可能是肺损伤的一个致病因素，我们分离了对苯醌，并将其病理生理作用与香烟烟雾进行了比较。由于维生素C是一种强抗氧化剂，我们还确定了维生素C对预防病理生理事件的调节作用。维生素C限制性豚鼠暴露于香烟烟雾（5支/天; 2喷/支）21天，补充和不补充15 mg维生素C/豚鼠/天。在体外、离体A549细胞以及豚鼠体内评估氧化损伤、凋亡和肺损伤。通过BALF中的嗜中性粒细胞来测量炎症。从新鲜制备的香烟烟气水提液中分离得到对苯半醌，并通过各种物理化学方法，包括质谱、NMR和ESR光谱对其进行了表征。用豚鼠肺内滴注对苯半醌，观察对苯半醌所致的肺损伤。肺损伤通过增加的空气空间来测量，如通过组织学和形态测定分析所证明的。通过Western印迹分析测定氧化蛋白损伤、MMPs、VEGF和VEGFR2，并使用MALDI-TOF-MS测定Michael加合物的形成。通过TUNEL测定证实凋亡，使用免疫印迹分析和共聚焦显微镜观察caspase 3的活化、PARP的降解和Bax/Bcl-2比率的增加。豚鼠暴露于香烟烟雾导致进行性蛋白质损伤，炎症，细胞凋亡和肺损伤长达21天的实验期。给予15 mg维生素C/豚鼠/天可预防所有这些病理生理学效应。对苯醌模拟香烟烟雾在体外和离体A549细胞中引起蛋白质修饰和凋亡以及在体内引起凋亡和肺损伤。所有这些病理生理学事件也被维生素C预防。对苯半醌似乎是香烟烟雾诱导的氧化蛋白损伤的主要原因，导致细胞凋亡和肺损伤。病理生理学事件可通过中等剂量的维生素C预防。
Cigarette smoke-induced cellular and molecular mechanisms of lung injury are not clear. Cigarette smoke is a complex mixture containing long-lived radicals, including p-benzosemiquinone that causes oxidative damage. Earlier we had reported that oxidative protein damage is an initial event in smoke-induced lung injury. Considering that p-benzosemiquinone may be a causative factor of lung injury, we have isolated p-benzosemiquinone and compared its pathophysiological effects with cigarette smoke. Since vitamin C is a strong antioxidant, we have also determined the modulatory effect of vitamin C for preventing the pathophysiological events. Vitamin C-restricted guinea pigs were exposed to cigarette smoke (5 cigarettes/day; 2 puffs/cigarette) for 21 days with and without supplementation of 15 mg vitamin C/guinea pig/day. Oxidative damage, apoptosis and lung injury were assessed in vitro, ex vivo in A549 cells as well as in vivo in guinea pigs. Inflammation was measured by neutrophilia in BALF. p-Benzosemiquinone was isolated from freshly prepared aqueous extract of cigarette smoke and characterized by various physico-chemical methods, including mass, NMR and ESR spectroscopy. p-Benzosemiquinone-induced lung damage was examined by intratracheal instillation in guinea pigs. Lung damage was measured by increased air spaces, as evidenced by histology and morphometric analysis. Oxidative protein damage, MMPs, VEGF and VEGFR2 were measured by western blot analysis, and formation of Michael adducts using MALDI-TOF-MS. Apoptosis was evidenced by TUNEL assay, activation of caspase 3, degradation of PARP and increased Bax/Bcl-2 ratio using immunoblot analysis and confocal microscopy. Exposure of guinea pigs to cigarette smoke resulted in progressive protein damage, inflammation, apoptosis and lung injury up to 21 days of the experimental period. Administration of 15 mg of vitamin C/guinea pig/day prevented all these pathophysiological effects. p-Benzosemiquinone mimicked cigarette smoke in causing protein modification and apoptosis in vitro and in A549 cells ex vivo as well as apoptosis and lung damage in vivo. All these pathophysiological events were also prevented by vitamin C. p-Benzosemiquinone appears to be a major causative factor of cigarette smoke-induced oxidative protein damage that leads to apoptosis and lung injury. The pathophysiological events are prevented by a moderately large dose of vitamin C.
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影响因子：
5.8
作者：
Demedts, Ingel K.;Demoor, Tine;Bracke, Ken R.;Joos, Guy F.;Brusselle, Guy G.
通讯作者：
Brusselle, Guy G.
影响因子：
10
作者：
Churg, Andrew;Wang, Rona;Wright, Joanne L.
通讯作者：
Wright, Joanne L.
影响因子：
15.9
作者：
Kasahara, Y;Tuder, RM;Voelkel, NF
通讯作者：
Voelkel, NF
影响因子：
24.3
作者：
Imai, K;Mercer, BA;D'Armiento, JM
通讯作者：
D'Armiento, JM
影响因子：
24.3
作者：
Bartalesi, B;Cavarra, E;Lungarella, G
通讯作者：
Lungarella, G