PM2.5对大鼠哮喘形成及发展的影响及氢气的治疗效果

发布时间：2018-02-27 20:55

本文关键词： 细颗粒物氢气气道炎症氧化应激肺功能　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨PM2.5在大鼠哮喘形成中的作用以及吸入氢气对上述过程的干预效果。方法:1将35只wister雌性大鼠随机分配到5个组内,分别为空白对照组、哮喘模型组、哮喘模型+H2组、哮喘模型+PM 2.5组、哮喘模型+PM 2.5+H2组,采用卵清白蛋白、氢氧化铝干粉剂腹腔注射致敏及激发建立哮喘模型,于造模开始同时给与哮喘模型+PM 2.5组、哮喘模型+PM 2.5+H2组大鼠开始雾霾染毒,每天4小时,每周5天,暴露30天,吸入雾霾浓度控制于0.6mg/m3左右,同时,哮喘模型+PM 2.5+H2组大鼠于雾霾暴露完毕后同哮喘+氢气组哮喘大鼠一起给予氢气吸入治疗,暴露后吸入含66%H2的氢氧混合气,每天氢气治疗2小时,持续30天。2各组大鼠于雾霾暴露第30天结束时,应用3%戊巴比妥钠腹腔注射麻醉,气管切开,插管,应用肺功能仪检测各组大鼠在雾化吸入不同浓度乙酰甲胆碱后气道总阻力的变化;检测完气道阻力后,股动脉取血处死大鼠,血液室温放置,凝固后离心收集血清;打开胸腔,结扎右肺主支气管,取3mL PBS灌洗大鼠左肺,重复灌洗3次,离心后取上清,采用ELISA法检测大鼠血清和支气管肺泡灌洗液(BALF)中IL-13和OVA-sIgE水平;取大鼠右肺上叶肺组织于4%甲醛中固定48小时后,用于肺组织病理,于电子显微镜下观察各组大鼠肺组织病理结果。3数据分析:采用SPSS 21.0统计软件对所有数据进行统计分析。对于计量资料数据用均数土标准差来表示。检测数据正态性及正态方差齐的计量资料采用单因素方差分析(One-Way ANOVA),进一步两两比较均采用SNK法;以P0.05为具有统计学意义。结果:1大鼠雾化吸入0.9%氯化钠溶液后各组之间气道阻力无差异(P0.05);当吸入乙酰甲胆碱浓度为6.25mg/mL时,哮喘模型+PM 2.5组大鼠气道阻力明显高于其余各组,有统计学意义(P0.05),哮喘模型组大鼠气道阻力高于空白对照组但与哮喘模型+H2组、哮喘模型+PM 2.5+H2组之间无差异(P0.05);吸入乙酰甲胆碱浓度升至12.5mg/mL时,除哮喘模型+H2组与哮喘模型+PM 2.5+H2组无差异外,其余各组织间有明显差异。2各组血清及灌洗液中SIgE水平与空白组相比明显升高,两组之间对比有统计学差异(P0.05);同哮喘模型组大鼠相比,哮喘模型+H2组大鼠血清及肺泡灌洗液中SIg E水平明显降低,两组之间对比有统计学差异(P0.05);哮喘模型+PM 2.5同哮喘组大鼠相比较,哮喘模型+PM 2.5组大鼠炎症水平明显高于哮喘模型组大鼠,两组之间存在统计学差异(P0.05)。3对照组大鼠支气管管腔通畅,黏膜上皮完整一致。哮喘模型组大鼠支气管管腔狭窄;哮喘模型+PM2.5组大鼠支气管管腔狭窄程度较哮喘模型组明显加重;氢气治疗组大鼠其支气管管腔狭窄程度较哮喘模型组大鼠略有改善,但较空白对照组严重结论:雾霾可以加重大鼠气道反应性,可以增加气道对乙酰甲胆碱的敏感性,在哮喘形成过程中起到重要作用;氢气的抗炎作用,减轻实验大鼠气道炎症,一定程度上可以改善大鼠的气道高反应性。
[Abstract]:Objective: To investigate the PM2.5 formation in rat asthma and the effects of inhalation of hydrogen in the process of intervention. Methods: 1 35 Wister female rats were randomly divided into 5 groups, namely control group, asthma group, asthma model group +H2, model +PM 2.5 asthma group, asthma model +PM in group 2.5+H2, using egg albumin, aluminum hydroxide powder agent sensitized intraperitoneally and induced asthma model, from the start of the model at the same time give asthma model +PM 2.5 group, +PM 2.5+H2 group, asthma model rats began haze exposure, 4 hours a day, 5 days a week, 30 days of exposure, the concentration of haze control in about 0.6mg/m3, inhalation at the same time, asthma model of +PM rats in the 2.5+H2 group after exposure to haze with asthma + H2 group asthmatic rats given together with hydrogen gas inhalation treatment, after exposure to inhalation of hydrogen oxygen mixed gas containing 66%H2, every hydrogen treatment for 2 hours, for 30 days in.2 group The rats in the haze exposed for thirtieth days at the end of anesthesia, using 3% sodium pentobarbital intraperitoneal injection of tracheotomy, intubation, application of pulmonary function testing in rats inhaled different concentrations of methacholine airway resistance; airway resistance after test, blood from the femoral artery of rats, blood at room temperature serum was collected after solidification, centrifugal; open the chest, ligation of right main bronchus lung, 3mL PBS left lung lavage in rats, 3 repeated lavage after centrifugation, the supernatant, serum and bronchoalveolar lavage fluid of rats was detected by the method of ELISA (BALF) IL-13 and OVA-sIgE levels; and the right lung upper lobe lung in rats in 4% formaldehyde fixed after 48 hours for pulmonary pathology in the electron microscope.3 data analysis of lung tissue of rats pathological results: using SPSS 21 statistical software for statistical analysis of all data for the measurement data used. The number of soil are expressed as standard deviation. The detection data of normality and homogeneity of variance of the normal measurement data using single factor analysis of variance (One-Way ANOVA), a further 22 were compared by SNK method; the P0.05 was considered statistically significant. Results: no differences between the 1 rats inhaled 0.9% Sodium Chloride Solution group airway resistance (P0.05); when inhaled methacholine concentration is 6.25mg/mL, the airway resistance in asthma model group +PM 2.5 rats was significantly higher than that of other groups, with statistical significance (P0.05), airway resistance in asthma rats in model group were higher than those in blank control group and asthma model group +H2, the difference between the asthma model group 2.5+H2 +PM (P0.05); acetyl the concentration of methacholine inhalation to 12.5mg/mL, in addition to the asthma model in +H2 group and +PM 2.5+H2 group had no difference in asthma model, all groups were significantly different between.2 serum and lavage fluid SIgE levels compared with the control group in Ming Dynasty Between the two groups significantly increased, compared with statistical difference (P0.05); compared with the asthma group, asthma model group serum +H2 and SIg level of E rats by bronchoalveolar lavage fluid was significantly lower, compared between the two groups had significant difference (P0.05); asthma model +PM 2.5 rats compared with the asthmatic group, inflammation the level of asthma model of +PM rats in 2.5 groups were significantly higher than asthma model group rats, there were significant differences between the two groups (P0.05).3 control group rat bronchial lumen patency intact mucous membrane. The bronchial asthma model group rat luminal stenosis; the degree of asthma model rats in the +PM2.5 group of bronchial stenosis compared with the asthma model group was significantly increased hydrogen; rats in the treatment group and the degree of bronchial stenosis compared with the asthma model group rats were slightly improved, but compared with blank control group conclusion: severe haze can aggravate the airway responsiveness to methacholine airway, can increase Choline sensitivity plays an important role in the development of asthma. The anti-inflammatory effect of hydrogen can alleviate airway inflammation in rats, and can improve airway hyperresponsiveness to some extent.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R562.25

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