柴油尾气污染物诱导豚鼠咳嗽高敏感性及其气道炎症探讨

发布时间：2018-05-20 18:12

本文选题：柴油尾气污染物 + 咳嗽高敏感性　；参考：《广州医科大学》2017年硕士论文

【摘要】：机动车尾气排放是我国大城市空气污染的重要来源,PM2.5是重要的空气污染物。在交通相关空气污染物中,PM2.5主要来源于柴油机动车排放的废气。流行病学研究表明,交通相关空气污染与咳嗽的患病率密切相关。研究证实,迷走神经/气道的TRPA1能被柴油尾气等空气污染物激活,从而引起一系列呼吸道疾病。然而空气污染物诱发咳嗽的机制尚未明确,空气污染物与咳嗽敏感性的关系亦鲜有报道。本课题组前期进行了室外的豚鼠暴露实验,证实交通相关空气污染能诱发豚鼠咳嗽敏感性增高。然而,室外暴露存在较多干扰因素,暴露环境不可控、不稳定。因此,有必要构建可控的、稳定的室内污染物暴露实验,进一步探讨空气污染物与咳嗽敏感性的关系。因而,本研究通过构建可控的、稳定的柴油尾气室内暴露环境,观察柴油尾气污染物与豚鼠咳嗽敏感性的关系,探讨TRPA1在柴油尾气污染物影响豚鼠咳嗽敏感性当中的作用。目的构建可控、稳定的柴油尾气室内暴露诱导的豚鼠咳嗽高敏感模型并探讨其机制。方法1、动物分组:分为对照组、低浓度尾气暴露组、高浓度尾气暴露组、低浓度尾气暴露+HC-030031组、高浓度尾气暴露+HC-030031组。2、构建室内暴露环境:暴露组豚鼠放置于柴油尾气暴露室内进行暴露,每天3小时,连续14天。低浓度暴露组和低浓度尾气暴露+HC-030031组以PM2.5浓度为200μg/m3作为目标浓度,高浓度暴露组和高浓度尾气暴露+HC-030031组以PM2.5浓度为1000μg/m3作为目标浓度。对照组豚鼠饲养于相对洁净的实验动物房。3、药物干预:低浓度尾气暴露+HC-030031组和高浓度尾气暴露+HC-030031组的每只豚鼠给予TRPA1特异性拮抗剂HC-030031(200mg/kg,ip)。每天给药一次,于咳嗽激发前2h最后一次给药。其余各组豚鼠不做任何给药干预。4、咳嗽激发试验后12h处理动物,留取标本,进行BALF细胞总数计数和分类计数、肺组织形态学观察、ELISA法检测肺组织匀浆的P物质含量、Western Blot法分别检测迷走神经节匀浆和肺组织匀浆的TRPA1含量。结果1、柴油尾气暴露可使豚鼠的咳嗽潜伏期均缩短、咳嗽激发次数均增多,该效应随尾气浓度的增高而愈发明显。HC-030031可在一定程度上延长咳嗽潜伏期、减少咳嗽激发次数。2、柴油尾气暴露后,BALF细胞总数增多,且随尾气浓度的增高而增多。柴油尾气暴露可诱导豚鼠产生气道炎症,炎症细胞以中性粒细胞和嗜酸性粒细胞为主,炎症细胞比例随尾气浓度的增高而增高。HC-030031可使细胞总数、中性粒细胞和嗜酸性粒细胞比例下降。3、柴油尾气暴露后,暴露组肺组织出现不同程度的血管/支气管周围炎、组织水肿和支气管上皮脱落。HC-030031可在一定程度上抑制该效应。4、柴油尾气暴露后,肺组织SP含量增多,且随暴露浓度的增高而增多;HC-030031可减少肺组织SP含量。5、柴油尾气暴露后,肺组织、迷走神经节的TRPA1含量均有不同程度的增多。两个迷走神经节中,TRPA1含量增多以颈静脉神经节为主。HC-030031可减少肺组织、迷走神经节的TRPA1含量。结论:1、本实验构建之柴油尾气室内暴露环境可诱导豚鼠咳嗽敏感性增高,产生气道炎症,该暴露方法可控、稳定,可作为研究咳嗽高敏感性的一种动物模型。2、柴油尾气污染物可诱导豚鼠产生非特异性气道炎症,以中性粒细胞和嗜酸性粒细胞增多为主,同时产生以P物质增多的气道神经源性炎症。3、柴油尾气污染物可诱导豚鼠出现咳嗽高敏感性,与感觉神经/气道TRPA1的激活有关。
[Abstract]:Vehicle exhaust emission is an important source of air pollution in large cities in China. PM2.5 is an important air pollutant. In traffic related air pollutants, PM2.5 is mainly derived from exhaust emissions from diesel motor vehicles. Epidemiological studies have shown that traffic related air pollution is closely related to the incidence of coughing. Research has confirmed that vagus nerve / gas TRPA1 can be activated by air pollutants such as diesel exhaust, causing a series of respiratory diseases. However, the mechanism of air pollutants induced coughing is not yet clear, and the relationship between air pollutants and cough sensitivity is rarely reported. The cough sensitivity of guinea pigs increased. However, there were many disturbing factors in outdoor exposure, and the exposure environment was uncontrollable and unstable. Therefore, it is necessary to construct a controlled, stable indoor pollutant exposure experiment, and further explore the relationship between air pollutants and cough sensitivity. Therefore, this study is based on the construction of a controlled, stable diesel exhaust indoor storm. The relationship between the pollutants of diesel exhaust and the cough sensitivity of guinea pigs was observed, and the role of TRPA1 in the coughing sensitivity of the diesel exhaust in guinea pigs was investigated. Objective to construct a controllable and stable high sensitivity model of the guinea pig cough induced by diesel exhaust indoor exposure and explore its mechanism. Methods 1, the animals were divided into control group and low concentration group. The tail gas exposure group, the high concentration tail gas exposure group, the low concentration tail gas exposure +HC-030031 group, the high concentration tail gas exposing the +HC-030031 group.2 and the indoor exposure environment: the exposed group guinea pigs were exposed to the diesel exhaust exposure room for 3 hours a day for 14 days. The low concentration exposed group and the low concentration tail gas exposed +HC-030031 group were PM2.5 concentration. The target concentration was 200 mu g/m3, the high concentration exposure group and the high concentration tail gas exposed the +HC-030031 group with the concentration of PM2.5 1000 g/m3 as the target concentration. The guinea pigs in the control group were raised in the relatively clean laboratory animal room.3, and the drug intervention was given to each guinea pig in the low concentration tail gas exposure +HC-030031 group and the high concentration tail gas exposed group +HC-030031 group to TRPA1. The specific antagonist HC-030031 (200mg/kg, IP) was given once a day and the last dose of 2H before the cough was stimulated. The rest of the guinea pigs were not given any medication to interfere with.4. After the cough stimulation test, the animals were treated with 12h, and the specimens were collected, the total number of BALF cells was counted and classified, the lung histomorphology observation, and the ELISA method were used to detect P in the lung tissue homogenate. Western Blot method was used to detect the TRPA1 content of the vagus ganglion homogenate and lung homogenate. Results 1, the exposure of diesel exhaust could shorten the cough latency and increase the number of coughing times in guinea pigs. The effect was increased with the increase of tail gas concentration and.HC-030031 could extend the latent period of coughing to a certain extent and reduce the cough. After the emission of.2, the total number of BALF cells increased and increased with the increase of tail gas concentration. The exposure of diesel exhaust could induce airway inflammation in guinea pigs. The inflammatory cells were mainly neutrophils and eosinophils. The proportion of inflammatory cells increased with the increase of tail gas concentration and.HC-030031 could make the total number of cells and neutral particles fine. The proportion of cell and eosinophil decreased by.3. After the exposure of diesel exhaust, the lung tissues of the exposed group showed different degree of blood vessel / bronchi, tissue edema and bronchial epithelium shedding of.HC-030031 could inhibit the effect of.4 to a certain extent. After the diesel exhaust was exposed, the SP content of lung tissue increased, and increased with the increase of exposure concentration; HC-03 0031 could reduce the SP content of lung tissue.5. After the diesel exhaust was exposed, the TRPA1 content of the lung tissue and the vagus ganglion increased in varying degrees. In the two vagus ganglion nodes, the increase of the content of TRPA1 in the jugular ganglion based.HC-030031 could reduce the TRPA1 content of the lung tissue and the vagus nerve node. Conclusion: 1, the diesel exhaust indoor storm was constructed in this experiment. Exposure environment can induce increased cough sensitivity and airway inflammation in guinea pigs. The exposure method is controllable and stable. It can be used as an animal model.2 for the study of cough Gao Min. The diesel exhaust pollutants can induce nonspecific airway inflammation in guinea pigs, mainly with neutrophils and eosinophils, and the increase of P substance. The airway neurogenic inflammation.3, diesel exhaust pollutants can induce cough in guinea pigs, Gao Min sense, and the activation of sensory nerve / airway TRPA1.
【学位授予单位】：广州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R56

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