维生素D2对哮喘大鼠肺组织IL-4及ICAM-1的影响

发布时间：2018-04-20 12:33

本文选题：维生素D2 + 哮喘　；参考：《安徽医科大学》2012年硕士论文

【摘要】：目的：哮喘是一种由多种细胞特别是肥大细胞、嗜酸性粒细胞、T淋巴细胞、嗜中性粒细胞、气道上皮细胞及多种细胞组分参与的慢性气道炎症反应性疾病，免疫调节在哮喘发病过程中有重要作用，维生素D在免疫调节中的作用越来越受到重视，研究发现维生素D对Th1/Th2平衡有调节作用，本实验通过维生素D2对哮喘大鼠模型进行干预，比较各组试验大鼠肺泡灌洗液中IL-4水平、肺泡灌洗液中细胞计数及分类、肺部ICAM-1的表达，探讨维生素D在哮喘大鼠的抗炎作用及其可能机制。方法：按照随机分组的原则，将30只SD雄性大鼠随机分组成3组，分别为哮喘组、维生素D2组、对照组，每组10只。（1）哮喘组：于第1、8、15天分别给予无菌腹腔注射10%OVA1ml（内含卵蛋白100mg及氢氧化铝100mg）致敏，自第16天开始置于密闭雾化罐中用1%OVA雾化吸入激发，持续雾化1周，每天激发30分钟致其出现呼吸急促、喘息或呼吸困难。（2）维生素D2(VD2)组：诱喘方法同哮喘组，从第15天开始给予腹腔注射维生素D25×10~5U/（kg.d），隔天1次，，共3次。（3）对照组：以生理盐水代替OVA和维生素D2。在末次激发后24小时内处死大鼠，收集肺组织及肺泡灌洗液（BALF）。将BALF离心后分离上清及沉淀细胞，并将肺组织经脱水透明、石蜡包埋等处理后制作成肺标本切片。运用Elisa方法对BALF上清中白细胞介素4（IL-4）水平进行测定，以及用改良牛氏计数台对BALF沉淀细胞进行计数并将沉淀细胞涂片染色后进行分类计数。运用免疫组织化学方法技术测定大鼠肺组织标本细胞间粘附分子1（ICAM-1）水平。通过统计学方法对各组指标数据进行分析比较。结果：VD2组IL-4含量（501.54±17.96）pg/ml明显低于哮喘组（615.37±94.77）pg/ml（P0.01）；VD2组IL-4含量与对照组比较无统计学意义；VD2组大鼠肺泡灌洗液沉渣中细胞总数（72.80±24.44）×10~6、中性粒细胞计数（18.20±8.14）×10~6、嗜酸性粒细胞计数（3.80±1.75）×10~6明显低于哮喘组大鼠肺泡灌洗液沉渣中细胞总数（10~8.92±27.55）×10~6、中性粒细胞计数（31.30±6.00）×10~6、嗜酸性粒细胞计数（7.10~±2.89）×10~6（P0.01）；对照组大鼠肺泡灌洗液沉渣中细胞总数（47.20±18.23）×10~6、中性粒细胞计数（7.40±1.96）×10~6、嗜酸性粒细胞计数（0.80±0.79）×10~6低于VD2组（P0.01）；VD2组肺组织ICAM-1表达（0.23±0.03）较哮喘组（0.34±0.10~）明显降低（P0.01）；VD2组大鼠肺组织ICAM-1表达（0.23±0.03）高于对照组（0.17±0.38）（P0.01）。结论：维生素D2能抑制Th2细胞因子IL-4水平，抑制哮喘大鼠气道ICAM-1水平，减少肺内气道炎症细胞浸润，减轻哮喘气道炎症反应。
[Abstract]:Objective: asthma is a chronic airway inflammatory response disease involving a variety of cells, especially mast cells, eosinophil T lymphocytes, neutrophilic granulocytes, airway epithelial cells and various cellular components. Immune regulation plays an important role in the pathogenesis of asthma. Vitamin D plays a more and more important role in the immune regulation. It has been found that vitamin D can regulate the balance of Th1/Th2. In this experiment, vitamin D _ 2 was used to interfere with asthma rat model. The levels of IL-4 in alveolar lavage fluid, cell count and classification in alveolar lavage fluid, and the expression of ICAM-1 in lung were compared in each group. To investigate the anti-inflammatory effect of vitamin D in asthmatic rats and its possible mechanism. Methods: according to the principle of random grouping, 30 SD male rats were randomly divided into three groups: asthma group, vitamin D2 group and control group. Asthma group (n = 10) was sensitized by intraperitoneal injection of 10OVA1ml (containing ovalbumin 100mg and 100mg aluminum hydroxide) on the 15th day of the first day, and was stimulated by 1%OVA atomization inhalation in a closed atomizing tank on the 16th day, and continued to atomize for 1 week. After 30 minutes of arousing every day, they developed shortness of breath, wheezing or dyspnea) vitamin D2 VD2) group: the asthma group received intraperitoneal injection of vitamin D 25 脳 10 5 U / L 路kg 路dL from the 15th day, once every other day. The control group was treated with normal saline instead of OVA and vitamin D 2. The rats were killed within 24 hours after the last stimulation, and the lung tissue and alveolar lavage fluid were collected. Supernatant and precipitate cells were isolated from BALF after centrifugation, and lung tissue was treated with dehydration and translucent, paraffin embedded. The level of IL-4 in the supernatant of BALF was determined by Elisa method, and the precipitation cells of BALF were counted by modified Niu's counter and stained by smear. The level of intercellular adhesion molecule (ICAM 1) in rat lung tissue was determined by immunohistochemical method. The index data of each group were analyzed and compared by statistical method. Results the IL-4 content in the VD2 group was significantly lower than that in the asthmatic group (615.37 卤94.77pgml / ml) and the IL-4 content in the VD2 group was significantly lower than that in the control group (72.80 卤24.44) 脳 10 ~ (-6), the neutrophil count was 18.20 卤8.14 脳 10 ~ (-6), and the eosinophil count was 3.80 卤1.75 脳 10 ~ (-6). The expression of ICAM-1 in lung tissue of VD2 group was significantly lower than that of VD2 group (0.23 卤0.03), which was significantly lower than that of asthmatic group (0.34 卤0.10). The expression of ICAM-1 in lung tissue of VD2 group was significantly lower than that of control group (0.17 卤0.38). Conclusion: vitamin D 2 can inhibit the level of Th2 cytokine IL-4, inhibit the level of airway ICAM-1 in asthmatic rats, reduce the infiltration of airway inflammatory cells in the lung, and alleviate the airway inflammation of asthma.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.6

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