辛伐他汀对哮喘小鼠气道黏液高分泌的影响

发布时间：2018-09-12 08:17

【摘要】：目的：观察哮喘小鼠气道炎症及气道黏液分泌情况。方法：清洁级BALB/c小鼠20只随机分为2组：哮喘组(AS组)10只,生理盐水对照组(NS组)10只。哮喘组以卵清白蛋白(OVA)致敏及激发制作哮喘模型,生理盐水对照组以生理盐水代替OVA,余处理同哮喘组。支气管肺泡灌洗液(BALF)用作细胞总数及细胞分类计数,ELISA检测支气管肺泡灌洗液中IL-5及IL-32水平,HE染色观察小鼠气道及肺组织病理变化,阿尔辛蓝-过碘酸雪夫染色(AB-PAS)观察气道组织杯状细胞、黏液分泌情况,免疫组织化学法(IHC)检测黏蛋白5AC(MUC5AC)在气道及肺组织分布情况,实时荧光定量PCR、蛋白免疫印迹法(Western blot)分别检测MUC5AC mRNA、MUC5AC蛋白在气道组织表达。结果：AS组小鼠BALF细胞总数、中性粒细胞数、淋巴细胞及嗜酸性粒细胞水平均高于NS组(P0.05),BALF中IL-5、IL-32水平、气道组织AB-PAS阳性染色面积、MUC5AC蛋白及mRNA表达均高于NS组(P0.05)。结论：哮喘小鼠存在气道炎症、气道黏液高分泌。目的：观察辛伐他汀对哮喘小鼠气道黏液高分泌的影响,探讨辛伐他汀在哮喘小鼠气道组织MUC5AC表达中作用机制。方法：将清洁级BALB/c小鼠50只随机分成5组：生理盐水对照组(NS组)、哮喘组(AS组)、辛伐他汀干预组(SIM组)、溶剂对照组(ET组)、地塞米松组(DEX组),每组10只。进行BALF细胞总数、白细胞分类计数,ELISA检测BALF中IL-5、IL-32水平,HE染色观察小鼠肺及气道组织病理变化,AB-PAS染色观察气道组织杯状细胞、黏液分泌情况,IHC、Western blot、实时荧光定量PCR检测MUC5AC蛋白及mRNA在气道及肺组织的表达。结果：AS组BALF细胞总数、中性粒细胞、淋巴细胞及嗜酸性粒细胞数、IL-5及IL-32水平、AB-PAS阳性染色面积、MUC5AC mRNA及蛋白水平、MUC5AC积分光密度值均较NS组、SIM组、DEX组增高,差异有统计学意义(P0.05)；与ET组相比,差异无统计学意义(P0.05)；DEX组、SIM组间差异无统计学意义(P0.05)。结论：辛伐他汀可抑制哮喘气道炎症,并且降低气道粘蛋白MUC5AC高分泌。
[Abstract]:Objective: to observe airway inflammation and airway mucus secretion in asthmatic mice. Methods: twenty clean grade BALB/c mice were randomly divided into two groups: asthma group (AS group, n = 10) and saline control group (NS group, n = 10). The asthmatic model was induced by ovalbumin (OVA) in asthmatic group. The control group was treated with saline instead of OVA,. Bronchoalveolar lavage fluid (BALF) was used as the count of total cells and cell classification. The levels of IL-5 and IL-32 in bronchoalveolar lavage fluid (BALF) were detected by Elisa and HE staining was used to observe the pathological changes of airway and lung tissue in mice. The goblet cells and mucus secretion of airway tissue were observed by AB-PAS, and the distribution of mucin 5AC (MUC5AC) in airway and lung tissue was detected by immunohistochemical (IHC). The expression of MUC5AC mRNA,MUC5AC protein in airway tissue was detected by real-time quantitative PCR, Western blot (Western blot). Results the total number of BALF cells, the number of neutrophils, the level of lymphocyte and eosinophil in as group were higher than those in NS group (P0.05), and the expression of MUC5AC protein and mRNA in AB-PAS positive staining area of airway tissue were higher than those in NS group (P0.05). Conclusion: asthma mice have airway inflammation and airway mucus secretion. Aim: to observe the effect of simvastatin on airway mucus hypersecretion in asthmatic mice and to explore the mechanism of simvastatin on the expression of MUC5AC in airway tissue of asthmatic mice. Methods: fifty BALB/c mice of clean grade were randomly divided into 5 groups: normal saline control group (NS group), asthma group (AS group), simvastatin intervention group (SIM group), solvent control group (ET group) and dexamethasone group (DEX group) with 10 rats in each group. The total number of BALF cells and the level of IL-5,IL-32 in BALF were detected by Elisa. The pathological changes of lung and airway tissues in mice were observed by HE staining. The goblet cells in airway tissue were observed by AB-PAS staining. The expression of MUC5AC protein and mRNA in airway and lung tissues were detected by real-time fluorescence quantitative PCR (PCR). Results the total number of BALF cells, the number of neutrophils, lymphocytes and eosinophilic granulocytes and the levels of AB-PAS positive staining area of MUC5AC mRNA and protein in as group were significantly higher than those in NS group (P0.05). Compared with the ET group, the difference was not statistically significant (P0.05). Conclusion: simvastatin can inhibit airway inflammation and reduce airway mucin MUC5AC hypersecretion.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R562.25

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