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目的:观察咪喹莫特对哮喘小鼠气道反应性,气道重塑和肺组织血管内皮生长因子(VEGF)表达的影响。方法:30只小鼠按随机数字表法分成3组,每组10只。正常对照组、哮喘组、咪喹莫特组。小鼠于第0、14天以鸡卵白蛋白(OVA)致敏,第24天开始雾化吸入1%OVA激发并持续28天,建立哮喘气道重塑模型,咪喹莫特组在吸入OVA前2h雾化吸入咪喹莫特30min。于最后一次雾化结束后24h,利用肺功能仪测小鼠气道阻力;收集支气管肺泡灌洗液(BALF)进行细胞计数及分类;对肺组织切片行HE染色观察病理学改变;运用医学图像分析软件测定肺组织切片中的血管计数、血管壁平滑肌厚度、血管壁平滑肌细胞计数;用免疫组化方法检测肺组织VEGF的蛋白表达水平;用RT-PCR检测肺组织VEGF mRNA表达水平。结果:哮喘组小鼠呼气阻力(Re)高于对照组(P<0.05),咪喹莫特组Re低于哮喘组(P<0.05);哮喘组BALF中细胞总数及各种炎症细胞数均较对照组升高(P<0.05),咪喹莫特组较哮喘组降低(P<0.05);哮喘组血管计数、血管壁平滑肌细胞计数较对照组增高,差异均有统计学意义(P<0.05),咪喹莫特组血管计数较哮喘组下降,差异有统计学意义(P<0.05);对照组VEGF蛋白和mRNA在气道不表达或轻度表达,哮喘组VEGF蛋白和mRNA表达较对照组增加,差异有统计学意义(P<0.05),咪喹莫特组能减少VEGF蛋白和mRNA的表达,与哮喘组比较差异有统计学意义(P<0.05),但较对照组仍增加,差异无统计学意义(P>0.05)。结论:早期预防性雾化吸入咪喹莫特通过部分抑制哮喘肺组织VEGF蛋白和mRNA的过度表达,阻止慢性哮喘小鼠的血管生成,可在一定程度上减轻哮喘小鼠的气道重塑和气道高反应性。
Objective: To observe the effect of imiquimod on the airway responsiveness, airway remodeling and the expression of vascular endothelial growth factor (VEGF) in asthmatic mice. Methods: Thirty mice were divided into three groups according to random number table method, with 10 mice in each group. Normal control group, asthma group and imiquimod group. Mice were sensitized with chicken ovalbumin (OVA) on day 0 and day 14, and aerosolized 1% OVA was started on day 24 for 28 days. Airway remodeling was established in the imiquimod group. OVA Inhalation of imiquimod 30 min before inhalation 2h. The airway resistance of mice was measured by pulmonary function 24h after the last atomization. The bronchoalveolar lavage fluid (BALF) was collected for cell counting and classification. The lung tissue sections were observed for pathological changes by HE staining. The expression of VEGF protein in lung tissue was detected by immunohistochemical method. The expression of VEGF mRNA in lung tissue was detected by RT-PCR. Results: The expiratory resistance (Re) of asthmatic mice was significantly higher than that of the control group (P <0.05), while that of the imiquimod group was lower than that of the asthmatic group (P <0.05). The total number of BALF and the number of inflammatory cells (P <0.05). The imiquimod group was lower than the asthma group (P <0.05). The numbers of vascular and vascular smooth muscle cells in the asthma group were significantly higher than those in the control group (P <0.05), and the difference was statistically significant (P (P <0.05). The blood counts of imiquimod group were lower than those of asthma group (P <0.05). The expression of VEGF protein and mRNA in the control group were either not expressed or mildly expressed in the airway, and the expression of VEGF protein and mRNA in the asthmatic group Compared with the control group, the difference was statistically significant (P <0.05), imiquimod group can reduce the expression of VEGF protein and mRNA, compared with the asthma group, the difference was statistically significant (P <0.05), but compared with the control group Increase, the difference was not statistically significant (P> 0.05). CONCLUSION: Early prophylactic nebulization of imiquimod inhibits the angiogenesis in chronic asthmatic mice by partially inhibiting the overexpression of VEGF protein and mRNA in asthmatic lung tissue, which may reduce the airway remodeling and gas remodeling in asthmatic mice to a certain extent Road hyperresponsiveness.