RNAi技术沉默STAT1基因对哮喘小鼠气道炎症的影响  

Effects of RNAi silencing STAT1 gene on airway inflammation of asthmatic mice

作　　者：刘春凤[1] 任磊[2] 熊瑛[1] 

机构地区：[1]泸州医学院附属医院呼吸内科,四川泸州646000 [2]泸州医学院附属医院胃肠外科,四川泸州646000

出　　处：《泸州医学院学报》2015年第1期7-10,共4页Journal of Luzhou Medical College

基　　金：四川省科技厅基金(编号:07104)

摘　　要：目的:探讨RNA干扰(RNA interference,RNAi)技术沉默转导子与转录活化子1(signal transducer and activator of transcription 1,STAT1)基因对支气管哮喘小鼠气道炎症的影响。方法 :将32只4周龄BALB/c雌鼠随机分为正常组、PBS组、空质粒(随机片段RNAi)组和si RNA(p GSTAT1-2)组,利用RNAi沉默支气管哮喘小鼠中STAT1基因,采用免疫组织化学检测STAT1蛋白的表达;通过单肺灌洗留取肺泡灌洗液(BALF),应用酶联免疫吸附试验法(ELISA)检测气道炎症介质如γ干扰素(IFN-γ)和白细胞介素5(IL-5)的表达水平。结果 :si RNA组的IFN-γ浓度高于PBS组、空质粒组(P<0.05),而IL-5的表达水平低于PBS组、空质粒组(P<0.05),二者在PBS组和空质粒组组间均无差异(P>0.05);si RNA组BALF中嗜酸性粒细胞(EOS)数量和IL-5浓度较空质粒组和PBS组中均明显减低,而IFN-γ的浓度增高,IFN-γ与EOS呈负相关,IL-5与EOS呈正相关,si RNA组BALF中白细胞(WBC)数量较空质粒组和PBS组中均明显减低;STAT-1蛋白主要表达于气道上皮细胞,si RNA组STAT-1蛋白表达明显低于PBS组与空质粒组,正常组低于si RNA组、PBS组和空质粒组,有统计学意义(P<0.05)。结论 :si RNA能有效抑制哮喘小鼠气道上皮细胞STAT1蛋白的表达,减轻哮喘小鼠的气道炎症反应。Objective： To discuss the effects of RNA interference（RNAi） silencing the gene of signal transducer and activator of transcription 1（STAT1） on airway inflammation of bronchial asthmatic mice. Methods：4 weeks old female BALB/c from 32 mice were randomly divided into normal group, PBS group, empty plasmid（random fragment RNAi） group and si RNA（p GSTAT1-2） group. RNAi was used to silence the gene of STAT1 of bronchial asthmatic mice. S-P method was used to detect the expression of STAT1 and ELISA was used to detect the concentration of airway inflammation medium such as IFN-γ and IL-5 from the bronchoalveolar lavage fluid（BALF） by the single lung lavage. Results： The concentration of IFN-γ of si RNA group was higher than that of PBS group and empty plasmid group（P〈0.05）, but the concentration of IL-5 was opposite（P〈0.05）. There was no significant difference between PBS group and empty plasmid group（P〈0.05）. The count of EOS and the concentration of IL-5 were significantly decreased in empty plasmid group and the PBS group, but the concentration of IFN-γ increased, indicating negative correlation between IFN-γ and EOS, and positively correlation between IL-5 and EOS. The count of WBC in BALF of si RNA group was lower than that of PBS group and empty plasmid group（P〈0.05）. STAT-1 was expressed in airway epithelial cells, the expression of STAT-1of si RNA group was significantly lower than that of PBS group and empty plasmid group, while that of the normal group was the lowest（P〈0.05）. Conclusion： Si RNA could effectively inhibit the expression of STAT1 of airway epithelial cell and alleviate airway inflammation in asthmatic mice.

关 键 词：哮喘 沉默转导子与转录活化子1 RNA干扰. 

分 类 号：Q786[生物学—分子生物学]