烟雾暴露对卵白蛋白致敏的急性支气管哮喘大鼠模型肺组织激活素A mRNA和蛋白表达的影响

Effect of smoke exposure on the expression of activin A mRNA and protein in lung tissue of rats with acute asthma sensitized by ovalbumin

目的分析烟雾暴露对卵白蛋白致敏的急性支气管哮喘大鼠模型肺组织激活素A mRNA和蛋白表达的影响。方法取28只健康成年雄性SD大鼠,按照随机分配原则,分为空白对照组(正常大鼠,不采取任何处理)、阴性对照组(正常大鼠+烟雾暴露)、模型组(制备卵白蛋白致敏的急性支气管哮喘大鼠模型)、实验组(急性支气管哮喘大鼠+烟雾暴露),每组各7只。行苏木精-伊红染色,观察各组大鼠肺组织病理学改变;采用免疫印迹法检测大鼠肺组织中激活素A蛋白表达水平;根据实时荧光定量聚合酶链反应检测大鼠肺组织中激活素A mRNA相对表达量。结果经病理学染色结果可知,空白对照组肺组织病理学未见异常,而阴性对照组、模型组及实验组肺组织出现不同程度的病理学改变,且实验组更加明显。经免疫印迹法检测可知,阴性对照组、模型组及实验组肺组织中激活素A蛋白表达水平较空白对照组均明显升高(P <0. 05);模型组肺组织中激活素A蛋白表达水平较阴性对照组明显升高(P <0. 05);实验组肺组织中激活素A蛋白表达水平较阴性对照组和模型组明显升高(P <0. 05)。经实时荧光定量聚合酶链反应检测可知,相比空白对照组,阴性对照组、模型组及实验组肺组织中激活素A mRNA相对表达量均明显上调(P <0. 05);相比阴性对照组,模型组肺组织中激活素A mRNA相对表达量明显上调(P <0. 05);相比阴性对照组和模型组,实验组肺组织中激活素A mRNA相对表达量明显上调(P <0. 05)。结论在卵白蛋白致敏的急性支气管哮喘大鼠模型中,经烟雾暴露可促进大鼠肺组织激活素A蛋白表达,提高激活素A mRNA表达量,并进一步导致气道炎症加重。

Objective To analyze the effect of smoke exposure on the expression of activin A mRNA and protein in the lung tissue of rats with acute asthma sensitized by ovalbumin. Methods 28 healthy adult male SD rats were randomly divided into blank control group( normal rats,without any treatment),negative control group( normal rats + smoke exposure),model group( ovalbumin sensitized acute asthma rat model),experimental group( acute asthma rats + smoke exposure),7 rats in each group. Hematoxylin eosin staining was used to observe the pathological changes of lung tissue in each group;the expression level of activin A protein in lung tissue was detected by immunoblotting;the relative expression level of activin A mRNA in lung tissue was detected by real-time fluorescence quantitative PCR. Results The results of pathological staining showed that the lung histopathology of the blank control group was normal,while that of the negative control group,the model group and the experimental group showed pathological changes in different degrees,and the experimental group was more obvious. The results of western blot showed that the expression level of activin A protein in the lung tissue of the negative control group,model group and experimental group was significantly increased compared with the blank control group( P < 0. 05);the expression level of activin A protein in the lung tissue of the model group was significantly increased compared with the negative control group( P < 0. 05);the expression level of activin A protein in the lung tissue of the experimental group was significantly increased compared with the negative control group and model group( P < 0. 05). Compared with the blank control group,the relative expression of activin A mRNA in the lung tissue of the negative control group,the model group and the experimental group was significantly increased( P < 0. 05). Compared with the negative control group,the relative expression of activin A mRNA in the lung tissue of the model group was significantly increased( P < 0. 05);compared with the negative control group and the model group,the relative expression of activin A mRNA in the lung tissue of the experimental group was significantly increased( P < 0. 05). Conclusion In the model of acute asthma rats sensitized by ovalbumin,smoke exposure can promote the expression of activin A protein in lung tissue,increase the expression of activin A mRNA,and further lead to the aggravation of airway inflammation.