吸入脂多糖对哮喘小鼠气道炎症和气道黏液分泌的影响

发布时间：2018-06-17 23:40

本文选题：哮喘 + 小鼠　；参考：《四川大学学报(医学版)》2014年03期

【摘要】：目的观察哮喘小鼠吸入脂多糖(LPS,作为刺激原)其气道炎症和气道黏液分泌的变化。方法30只清洁级BALB/c小鼠随机分为哮喘组(AST组)、LPS哮喘组(LAS组)和正常组(NS组),每组10只。哮喘组用卵清白蛋白(OVA)致敏和激发制作哮喘模型,LPS哮喘组在哮喘模型的基础上加用LPS(50μg/mL)雾化吸入30min,正常组用生理盐水代替OVA。检测各组小鼠支气管肺泡灌洗液(BALF)细胞总数和细胞分类计数,采用ELISA检测BALF中的白细胞介素4(IL-4)和肿瘤坏死因子-α(TNF-α)水平,HE染色观察肺部病理学改变,用阿尔辛蓝-过碘酸雪夫(AB-PAS)染色气道杯状细胞,免疫组织化学法检测肺组织中黏蛋白5ac(Muc5ac)的表达,荧光定量RT-PCR检测Muc5ac mRNA在肺内的表达。并分析Muc5ac蛋白表达与各指标的相关性。结果AST及LAS组小鼠较NS组在BALF中的细胞总数、嗜酸性粒细胞、单核细胞、淋巴细胞百分比,IL-4和TNF-α水平、肺组织AB-PAS阳染面积,Muc5ac蛋白和mRNA表达明显升高,其差异均有统计学意义(P均0.05)。LAS组较AST组上述气道炎症(除单核细胞数外)和气道黏液高分泌指标明显增高,差异均有统计学意义(P0.05)。气道Muc5ac蛋白表达与BALF中细胞总数、嗜酸性粒细胞数、IL-4、TNF-α水平、气道AB-PAS染色阳性着色面积均呈正相关(P均0.05)。结论 OVA致敏和激发的哮喘小鼠出现以嗜酸性粒细胞、淋巴细胞浸润为主的气道炎症及杯状细胞增生的气道黏液高分泌,且气道炎症和气道黏液高分泌关系密切。LPS可使气道炎症和气道黏液高分泌加重,可能与LPS激发了体内炎症介质的生成、活化有关。
[Abstract]:Objective to observe the changes of airway inflammation and airway mucus secretion in asthmatic mice by inhaling lipopolysaccharide (LPS). Methods Thirty clean BALB / c mice were randomly divided into two groups: Asthma group (AST group) and LPS group (Las group) and normal control group (10 mice in each group). Asthmatic group was sensitized with ovalbumin (OVA) and stimulated to make asthmatic model. On the basis of asthma model, the asthmatic group was inhaled with 50 渭 g / mL LPSN for 30 min, and the normal group was replaced by normal saline instead of OVA. The total number of BALF cells and the number of BALF cells in BALF of each group were measured. The levels of IL-4 and TNF- 伪 in BALF were detected by Elisa and the pathological changes of lung were observed by HE staining. The airway goblet cells were stained with AlsinBlue and Scheffer periodate (AB-PAS). The expression of mucin 5acine Muc5ac. and the expression of Muc5ac mRNA in lung tissue were detected by immunohistochemistry and fluorescence quantitative RT-PCR. The correlation between Muc5ac protein expression and various indexes was analyzed. Results the total number of cells, the percentage of eosinophilic granulocytes, monocytes, lymphocyte percentage and the expression of Muc5ac protein and mRNA in lung tissue in AST and Las group were significantly higher than those in NS group. Compared with AST group, the airway inflammation (except monocyte number) and the hypersecretion index of airway mucus were significantly increased in both groups (P < 0.05). The expression of Muc5ac protein in airway was positively correlated with the total number of cells in BALF, the number of eosinophils and the level of IL-4 and TNF- 伪, as well as the positive staining area of AB-PAS staining in the airway (P < 0.05). Conclusion in OVA sensitized and stimulated asthmatic mice, airway mucus secreted mainly by eosinophilic granulocyte, lymphocyte infiltration and goblet cell proliferation. Airway inflammation and airway mucus hypersecretion were closely related. LPS could aggravate airway inflammation and airway mucus hypersecretion, which may be related to the production and activation of inflammatory mediators stimulated by LPS in vivo.
【作者单位】：四川省仁寿县人民医院ICU;遵义医学院附属医院呼吸二科;
【分类号】：R562.25

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