β_(2)-AR减敏哮喘小鼠模型的建立及验证

Establishment and verification of the β_(2)-AR desensitization asthma mice model

目的:建立β_(2)-AR减敏哮喘小鼠模型并对其进行验证。方法:SPF级雄性BALB/c30只小鼠随机分为空白组、普通哮喘模型组、β_(2)-AR减敏哮喘模型组。建立普通哮喘模型,并在此基础上采用雾化吸入同时腹腔注射沙丁胺醇的方法进行β_(2)-AR减敏哮喘模型的制备,造模21 d末次激发后,测定小鼠气道阻力、ELISA法检测小鼠血清IgE含量,HE染色观察肺组织炎细胞浸润程度,Western blot法检测肺组织中β_(2)-AR含量,RT-PCR检测肺组织中β_(2)-ARmRNA的表达。结果:与空白组相比,随着乙酰甲胆碱(Mch)浓度升高,OVA诱导的各组气道阻力升高,β_(2)-AR减敏哮喘模型组气道阻力增加更加显著(P<0.05);与空白组相比,普通哮喘组及β_(2)-AR减敏哮喘模型组IgE水平上升(P<0.01);病理组织学观察发现β_(2)-AR减敏哮喘小鼠气道炎症浸润,黏液过度分泌及胶原明显沉积,且均较普通哮喘模型组的病理表现显著加重;β_(2)-AR减敏哮喘小鼠模型肺组织中β_(2)-AR含量及β_(2)-ARmRNA的表达水平较空白组及普通哮喘模型组均明显下降(P<0.05)。结论:β_(2)-AR减敏哮喘小鼠模型构建成功,且造模周期短。

Objective:To establish and verify a β_(2)-AR desensitization asthma mice model.Methods:A total of 30 SPF male BALB/c mice were randomly divided into blank group,the common asthma group,and β_(2)-AR desensitization asthma model group.Asthma model was established,and on this basis,the method of atom-izing inhalation and intraperitoneal injections of salbutamol was used to prepare β_(2)-AR desensitization asthma model.After the last stimulation on the 21st day of modeling,the airway resistance of mice was measured.ELISA was used to detect the content of serum IgE;HE staining was used to observe the lung organization degree of infla-mmatory cell infiltration;Western blot method was used to detect the β_(2)-AR content in lung tissue,RT-PCR was used to detect the β_(2)-ARmRNA expressionin lung tissue.Results:Compared with the blank group,as acetyl choline(Mch)levels increased,groups of OVA induced airway resistance increases;but the β_(2)-AR desensitization asthma model group increased airway resistance was more significant(P<0.05);compared with the blank group,IgE levels of common asthma group and β_(2)-AR desensitization asthma model group elevated(P<0.01).The pathological histology observation found the β_(2)-AR desensitization asthma airway inflammation infiltration in mice,the excessive mucus secretion and collagen deposition,and the pathological performance obviously increase compared with the common asthma group;β_(2)-AR content in the lung tissue of β_(2)-AR desensitization asthma model in mice,β_(2)-AR mRNA expression level in the blank group and common asthma model group were significantly decreased(P<0.05).Conclusion:The β_(2)-AR desensitization asthma mouse model was successfully established,and the buildingcycle was short.