Alcohol potentiates RSV-mediated injury to ciliated airway epithelium.
Alcohol potentiates RSV-mediated injury to ciliated airway epithelium.
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酒精会增强 RSV 介导的纤毛气道上皮损伤。
DOI：
10.1016/j.alcohol.2018.07.010
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Sisson,JosephH
中科院分区：
文献类型：
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作者：
Wyatt,ToddA;Bailey,KristinaL;Simet,SamanthaM;Warren,KristiJ;Sweeter,JeneaM;DeVasure,JaneM;Pavlik,JaquelineA;Sisson,JosephH
酒精会损害呼吸道病毒感染的消退。许多免疫反应途径因酒精滥用而发生改变，包括酒精引起的肺部纤毛功能障碍。我们假设，酒精暴露会损害粘液纤毛清除介导的针对呼吸道合胞病毒（RSV）的先天免疫。使用 Sisson-Ammons 视频分析对气-液界面上生长的小鼠气管上皮细胞的多个全场测量中运动点的平均数量进行定量，从而对纤毛进行分析。单独用乙醇预处理（100 mM，24 小时）对运动纤毛的数量没有影响。单剂量 (TCID501 × 105) RSV 导致 2 天后运动纤毛显着减少 (p< 0.05)。乙醇预处理显着 (p< 0.05) 增强 RSV 诱导的纤毛损失 2 天。 RSV 和乙醇联合治疗导致 PKC epsilon (PKCε) 持续激活诱导的自动下调。动力蛋白 ELISA 和上清液的 LDH 活性证实了乙醇诱导的纤毛细胞脱离增强。 RSV 诱导的纤毛损失在 7 天之前都很明显，此时仅感染 RSV 的细胞与对照细胞相比没有表现出明显的纤毛损失。然而，用乙醇预处理的细胞直到 RSV 感染后 10 天才显示出显着的纤毛损失。为了阐明乙醇增强的纤毛脱离的功能意义，将随意喂食酒精（20%，持续 12 周）的小鼠用 RSV 感染一次，并通过肺匀浆的空斑形成测定法测量清除情况，持续时间长达 7 天。 3 天后，在对照小鼠的肺部不再检测到 RSV 斑块形成，而在酒精喂养的小鼠中，第 7 天检测到显着 (p< 0.01) RSV 斑块形成单位。根据野生型 C57BL/6 小鼠（而非 PKCε KO 小鼠）气管测量结果显示，酒精喂养的小鼠纤毛损失增加，纤毛恢复延迟。这些数据表明，酒精以 PKCε 依赖性方式加重 RSV 介导的纤毛上皮损伤。
Alcohol impairs resolution of respiratory viral infections. Numerous immune response pathways are altered in response to alcohol misuse, including alcohol-induced ciliary dysfunction in the lung. We hypothesized that mucociliary clearance-mediated innate immunity to respiratory syncytial virus (RSV) would be compromised by alcohol exposure. Cilia were assayed using Sisson-Ammons Video Analysis by quantitating the average number of motile points in multiple whole field measurements of mouse tracheal epithelial cells grown on an air-liquid interface. Pretreatment with ethanol alone (100 mM for 24 hours) had no effect on the number of motile cilia. A single dose (TCID501 × 105) of RSV resulted in a significant (p< 0.05) decrease in motile cilia after 2 days. Ethanol pretreatment significantly (p< 0.05) potentiated RSV-induced cilia loss by 2 days. Combined RSV and ethanol treatment led to a sustained activation-induced auto-downregulation of PKC epsilon (PKCε). Ethanol-induced enhancement of ciliated cell detachment was confirmed by dynein ELISA and LDH activity from the supernates. RSV-induced cilia loss was evident until 7 days, when RSV-only infected cells demonstrated no significant cilia loss vs. control cells. However, cells pretreated with ethanol showed significant cilia loss until 10 days post-RSV infection. To address the functional significance of ethanol-enhanced cilia detachment, mice fed alcoholad libitum(20% for 12 weeks) were infected once with RSV, and clearance was measured by plaque-forming assay from lung homogenates for up to 7 days. After 3 days, RSV plaque formation was no longer detected from the lungs of control mice, while significant (p< 0.01) RSV plaque-forming units were detected at 7 days in alcohol-fed mice. Alcohol-fed mice demonstrated enhanced cilia loss and delayed cilia recovery from tracheal measurements in wild-type C57BL/6 mice, but not PKCε KO mice. These data suggest that alcohol worsens RSV-mediated injury to ciliated epithelium in a PKCε-dependent manner.