The Th17/Treg Cytokine Imbalance in Chronic Obstructive Pulmonary Disease Exacerbation in an Animal Model of Cigarette Smoke Exposure and Lipopolysaccharide Challenge Association
The Th17/Treg Cytokine Imbalance in Chronic Obstructive Pulmonary Disease Exacerbation in an Animal Model of Cigarette Smoke Exposure and Lipopolysaccharide Challenge Association
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DOI：
10.1038/s41598-019-38600-z
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发表时间：
2019-02-13
影响因子：
4.6
通讯作者：
Lopes, Fernanda D. T. Q. S.
中科院分区：
文献类型：
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作者：
Cervilha, Daniela A. B.;Ito, Juliana T.;Lopes, Fernanda D. T. Q. S.
我们提出了一个实验模型来验证COPD急性加重期Th 17/Treg细胞因子失衡。将40只C57 BL/6小鼠暴露于室内空气或香烟烟雾（CS）（12 +/-1支香烟，每天两次，30分钟/暴露和5天/周），并接受盐水（50 μ l）或脂多糖（LPS）（1 mg/kg在50 μ l盐水中）的气管内滴注。我们分析了支气管肺泡灌洗液和肺的平均线性截距、上皮厚度和炎症特征。我们使用免疫组织化学评价巨噬细胞、中性粒细胞、CD 4（+）和CD 8（+）T细胞、Treg细胞、IL-10（+）和IL-17（+）细胞以及STAT-3、STAT-5、磷酸化-STAT 3和磷酸化-STAT 5水平，使用ELISA评价IL-17、IL-6、IL-10、INF-γ、CXCL 1和CXCL 2水平。研究表明，CS暴露和LPS攻击增加了中性粒细胞、巨噬细胞以及CD 4（+）和CD 8（+）T细胞的数量。同时暴露于CS/LPS加剧了这种反应和肺实质损伤。LPS组和对照组的T淋巴细胞和IL-17（+）细胞密度及IL-17和IL-6水平均明显升高，而IL-10水平仅在对照/LPS组升高。STAT-3、磷酸化-STAT 3、STAT-5和磷酸化-STAT 5（+）细胞数量的增加证实了IL-17（+）和Treg细胞数量的增加。这些研究结果表明，同时挑战CS和LPS加剧了炎症反应，并诱导肺泡实质中的弥漫性结构变化，其特征在于Th 17细胞因子释放的增加。尽管观察到Treg细胞分化，但在CS/LPS组中观察到的IL-10表达的缺乏和IL 10（+）细胞密度的降低表明，在该提出的模型中，未能释放该细胞因子在加剧的炎症反应中起关键作用。
We proposed an experimental model to verify the Th17/Treg cytokine imbalance in COPD exacerbation. Forty C57BL/6 mice were exposed to room air or cigarette smoke (CS) (12 +/- 1 cigarettes, twice a day, 30 min/exposure and 5 days/week) and received saline (50 mu l) or lipopolysaccharide (LPS) (1 mg/kg in 50 mu l of saline) intratracheal instillations. We analyzed the mean linear intercept, epithelial thickness and inflammatory profiles of the bronchoalveolar lavage fluid and lungs. We evaluated macrophages, neutrophils, CD4(+) and CD8(+) T cells, Treg cells, and IL-10(+) and IL-17(+) cells, as well as STAT-3, STAT-5, phospho-STAT3 and phospho-STAT5 levels using immunohistochemistry and IL-17, IL-6, IL-10, INF-gamma, CXCL1 and CXCL2 levels using ELISA. The study showed that CS exposure and LPS challenge increased the numbers of neutrophils, macrophages, and CD4(+) and CD8(+) T cells. Simultaneous exposure to CS/LPS intensified this response and lung parenchymal damage. The densities of Tregs and IL-17(+) cells and levels of IL-17 and IL-6 were increased in both LPS groups, while IL-10 level was only increased in the Control/LPS group. The increased numbers of STAT-3, phospho-STAT3, STAT-5 and phospho-STAT5(+) cells corroborated the increased numbers of IL-17(+) and Treg cells. These findings point to simultaneous challenge with CS and LPS exacerbated the inflammatory response and induced diffuse structural changes in the alveolar parenchyma characterized by an increase in Th17 cytokine release. Although the Treg cell differentiation was observed, the lack of IL-10 expression and the decrease in the density of IL10(+) cells observed in the CS/LPS group suggest that a failure to release this cytokine plays a pivotal role in the exacerbated inflammatory response in this proposed model.