Cigarette Smoke Alters Respiratory Syncytial Virus-Induced Apoptosis and Replication
Cigarette Smoke Alters Respiratory Syncytial Virus-Induced Apoptosis and Replication
复制标题
DOI：
10.1165/rcmb.2008-0131oc
复制
发表时间：
2009-08-01
影响因子：
6.4
通讯作者：
Hunninghake, Gary W.
中科院分区：
文献类型：
--
作者：
Groskreutz, Dayna J.;Monick, Martha M.;Hunninghake, Gary W.
暴露于香烟烟雾的个体比不吸烟者有更多的病毒感染，包括呼吸道合胞病毒（RSV）感染，但细胞机制尚不清楚。我们的目的是确定香烟烟雾增强病毒感染的机制。我们假设香烟烟雾导致坏死并阻止病毒诱导的细胞凋亡，这与炎症和病毒复制增加有关。将原代气道上皮细胞暴露于香烟烟雾提取物2天，然后暴露于RSV 1天。Western blot检测切割的半胱天冬酶3和7显示，当细胞在病毒感染前用香烟烟雾处理时，细胞凋亡较少。细胞凋亡的ELISA和TUNEL检测证实了这一发现。测量细胞活力，包括碘化丙啶染色，ATP测定和细胞计数，表明香烟烟雾引起坏死，而不是病毒诱导的细胞凋亡。使用空斑试验和荧光标记的RSV，我们发现，虽然有香烟烟雾预处理组的活细胞较少，病毒载量增加。预处理的细胞与N-乙酰半胱氨酸和醛脱氢酶的效果被抑制，表明该效果主要是由反应性醛介导的。在病毒感染中，香烟烟雾引起坏死而不是凋亡，导致炎症增加和病毒复制增强。
Individuals exposed to cigarette smoke have a greater number and severity of viral infections, including respiratory syncytial virus (RSV) infections, than do nonsmokers, but the cellular mechanism is unknown. Our objective was to determine the mechanism by which cigarette smoke augments viral infection. We hypothesize that cigarette smoke causes necrosis and prevents virus-induced cellular apoptosis, and that this is associated with increased inflammation and viral replication. Primary airway epithelial cells were exposed to cigarette smoke extract for 2 days, followed by 1 day of RSV exposure. Western blot detection of cleaved caspases 3 and 7 showed less apoptosis when cells were treated with cigarette smoke before viral infection. This finding was confirmed with ELISA and TUNEL detection of apoptosis. Measures of cell viability, including propidium iodide staining, ATP assay, and cell counts, indicated that cigarette smoke causes necrosis rather than virus-induced apoptosis. Using plaque assay and fluorescently-labeled RSV, we showed that although there were less live cells in the cigarette smoke-pretreated group, viral load was increased. The effect was inhibited by pretreatment of cells with N-acetylcysteine and aldehyde dehydrogenase, suggesting that the effect was primarily mediated by reactive aldehydes. Cigarette smoke causes necrosis rather than apoptosis in viral infection, resulting in increased inflammation and enhanced viral replication.