Poole Jill A, Romberger Debra J, Bauer Chris, Gleason Angela M, Sisson Joseph H, Oldenburg Peter J, West William W, Wyatt Todd A
Pulmonary, Critical Care, Sleep and Allergy Division, Department of Internal Medicine, University of Nebraska Medical Center, Omaha, Nebraska 68198-5300, USA.
Exp Lung Res. 2012 Oct;38(8):383-95. doi: 10.3109/01902148.2012.714841. Epub 2012 Aug 16.
Organic dust samples from swine confinement facilities elicit pro-inflammatory cytokine/chemokine release from bronchial epithelial cells and monocytes, dependent, in part, upon dust-induced activation of the protein kinase C (PKC) isoform, PKCε. PKCε is also rapidly activated in murine tracheal epithelial cells following in vivo organic dust challenges, yet the functional role of PKCε in modulating dust-induced airway inflammatory outcomes is not defined. Utilizing an established intranasal inhalation animal model, experiments investigated the biologic and physiologic responses following organic dust extract (ODE) treatments in wild-type (WT) and PKCε knock-out (KO) mice. We found that neutrophil influx increased more than twofold in PKCε KO mice following both a one-time challenge and 3 weeks of daily challenges with ODE as compared with WT mice. Lung pathology revealed increased bronchiolar and alveolar inflammation, lymphoid aggregates, and T cell influx in ODE-treated PKCε KO mice. Airway hyperresponsiveness to methacholine increased in PKCε KO + ODE to a greater magnitude than WT + ODE animals. There were no significant differences in cytokine/chemokine release elicited by ODE treatment between groups. However, ODE-induced nitric oxide (NO) production differed in that ODE exposure increased nitrate levels in WT mice but not in PKCε KO mice. Moreover, ODE failed to upregulate NO from ex vivo stimulated PKCε KO lung macrophages. Collectively, these studies demonstrate that PKCε-deficient mice were hypersensitive to organic dust exposure and suggest that PKCε is important in the normative lung inflammatory response to ODE. Dampening of ODE-induced NO may contribute to these enhanced inflammatory findings.
来自猪舍设施的有机粉尘样本可促使支气管上皮细胞和单核细胞释放促炎细胞因子/趋化因子,这部分依赖于粉尘诱导的蛋白激酶C(PKC)同工型PKCε的激活。在体内有机粉尘刺激后,小鼠气管上皮细胞中的PKCε也会迅速激活,但PKCε在调节粉尘诱导的气道炎症结果中的功能作用尚未明确。利用已建立的鼻内吸入动物模型,实验研究了野生型(WT)和PKCε基因敲除(KO)小鼠在接受有机粉尘提取物(ODE)处理后的生物学和生理反应。我们发现,与WT小鼠相比,PKCε KO小鼠在一次性接受ODE刺激以及连续3周每日接受ODE刺激后,中性粒细胞流入增加了两倍多。肺部病理学显示,在接受ODE处理的PKCε KO小鼠中,细支气管和肺泡炎症、淋巴聚集物以及T细胞流入增加。与WT + ODE动物相比,PKCε KO + ODE小鼠对乙酰甲胆碱的气道高反应性增加幅度更大。两组之间ODE处理引发的细胞因子/趋化因子释放没有显著差异。然而,ODE诱导的一氧化氮(NO)产生有所不同,即ODE暴露使WT小鼠的硝酸盐水平升高,但PKCε KO小鼠则不然。此外,ODE未能上调离体刺激的PKCε KO肺巨噬细胞产生的NO。总体而言,这些研究表明,PKCε缺陷小鼠对有机粉尘暴露高度敏感,并提示PKCε在对ODE的正常肺部炎症反应中很重要。ODE诱导的NO的抑制可能导致这些炎症增强的结果。