Involvement of EGF receptor signaling and NLRP12 inflammasome in fine particulate matter-induced lung inflammation in mice
Involvement of EGF receptor signaling and NLRP12 inflammasome in fine particulate matter-induced lung inflammation in mice
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EGF 受体信号传导和 NLRP12 炎症小体参与细颗粒物诱导的小鼠肺部炎症
DOI：
10.1002/tox.22308
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发表时间：
2017-04-01
影响因子：
4.5
通讯作者：
Feng, Feifei
中科院分区：
文献类型：
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作者：
Jin, Yuefei;Wu, Weidong;Feng, Feifei
流行病学研究表明，暴露于环境细颗粒物（PM2.5）与呼吸道疾病有关。肺部炎症是许多肺部疾病的核心特征，PM2.5 暴露可诱发肺部炎症。然而，PM2.5 引起的肺部炎症的机制仍不清楚。为了表征表皮生长因子受体（EGFR）和炎症小体在 PM2.5 诱导的小鼠肺部炎症中的作用，连续 5 天分别向 30 只 BALB/c 小鼠支气管内滴注生理盐水和 PM2.5 悬浮液（4.0 mg/kg b.w.）。进行支气管肺泡灌洗（BAL）并收集BAL液（BALF）。使用 ELISA 测定 BALF 中活性氧 (ROS)、诱导型一氧化氮合酶 (iNOS)、表皮生长因子 (EGF)、CXCL1、白细胞介素 (IL)-1 和 IL-18 的水平。通过RT-PCR测定肺组织中IL-6、IL-1、IL-18、CXCL1、IL-10、NLRP3、Caspase-1和NLRP12的mRNA水平。分别使用免疫组织化学染色和蛋白质印迹法检查肺组织中的磷酸化 EGFR (Tyr1068) 和磷酸化 Akt (Thr308)。使用免疫组织化学染色检查支气管上皮中 Caspase-1、NLRP3、NF-B-p52/p100 和 NF-B-p65 的蛋白水平。研究表明，PM2.5 暴露会诱发肺部炎症。暴露于 PM2.5 的小鼠的总蛋白、ROS、iNOS、EGF 和 CXCL1 水平以及 BALF 中的细胞数量相对于对照组显着升高。与对照组相比，PM2.5暴露小鼠肺组织中CXCL1、IL-1和IL-18的mRNA水平增加。然而，PM2.5暴露小鼠肺组织中的NLRP12 mRNA水平降低。注入 PM2.5 的小鼠肺部的磷酸化 EGFR (Tyr1068) 和磷酸化 Akt (Thr308) 水平高于对照组小鼠的肺部。 PM2.5暴露小鼠支气管上皮中NF-B-p52/p100和NF-B-p65的蛋白水平也较对照组升高。这项研究表明 EGF-EGFR-Akt-NF-B 信号传导和 NLRP12 炎症小体可能与 PM2.5 诱导的小鼠肺部炎症有关。 (c) 2016 年 Wiley periodicals, Inc.Environ Toxicol 32：1121-1134，2017 年。
Epidemiological studies have shown that exposure to ambient fine particulate matter (PM2.5) is associated with respiratory diseases. Lung inflammation is a central feature of many pulmonary diseases, which can be induced by PM2.5 exposure. However, the mechanisms underlying PM2.5-induced lung inflammation remain unclear. To characterize the role of epidermal growth factor receptor (EGFR) and inflammasome in PM2.5-induced lung inflammation in mice, 30 BALB/c mice were intrabroncheally instilled with saline and PM2.5 suspension (4.0 mg/kg b.w.) for 5 consecutive days, respectively. Bronchoalveolar lavage (BAL) was conducted and BAL fluid (BALF) was collected. The levels of reactive oxygen species (ROS), inducible nitric oxide synthase (iNOS), epidermal growth factor (EGF), CXCL1, interleukin (IL)-1, and IL-18 in BALF were determined using ELISA. mRNA levels of IL-6, IL-1, IL-18, CXCL1, IL-10, NLRP3, Caspase-1, and NLRP12 in lung tissues were determined by RT-PCR. Phospho-EGFR (Tyr1068) and phospho-Akt (Thr308) in lung tissues were examined using immunohistochemical staining and Western blotting, respectively. Protein levels of Caspase-1, NLRP3, NF-B-p52/p100, and NF-B-p65 in bronchial epithelium were examined using immunohistochemical staining. It was shown that PM2.5 exposure induced lung inflammation. Levels of total protein, ROS, iNOS, EGF, and CXCL1 and cell number in the BALF of mice exposed to PM2.5 were markedly elevated relative to the control. mRNA levels of CXCL1, IL-1, and IL-18 in lung tissues of PM2.5-exposed mice were increased in comparison with the control. However, level of NLRP12 mRNA in lung tissues of PM2.5-exposed mice was reduced. Phospho-EGFR (Tyr1068) and phospho-Akt (Thr308) levels in the lungs of PM2.5-instilled mice were higher than those in the lungs of the control. The protein levels of NF-B-p52/p100 and NF-B-p65 in bronchial epithelium of PM2.5-exposed mice were also increased compared with the control. This study suggests that EGF-EGFR-Akt-NF-B signaling and NLRP12 inflammasome may be associated with PM2.5-induced lung inflammation in mice. (c) 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1121-1134, 2017.