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目的探究miR-27a对哮喘小鼠气道炎症的作用及机制。方法将雄性Balb/c小鼠随机分成正常对照组(control组)、哮喘模型组(OVA组)、哮喘+miR-27a模拟物组(miR-27a组)、哮喘+anti-miR-27a组(anti-miR-27a组)和哮喘+miR-27a对照组(scramble),每组8只。除正常对照组外,其余各组小鼠用卵清蛋白(OVA)致敏和激发建立哮喘模型;3组哮喘模型处理组通过鼻滴的方式从激发前1 d开始给予4 mg/kg相应的药物干预。实验第28天时检测气道高反应性和肺灌洗液(BALF)中细胞总数及分类计数,HE和PAS染色检测肺组织病理学变化,real-time PCR检测肺组织中miR-27a、NF-κB2、白细胞介素(interleukin,IL)-4、IL-13和干扰素(interferon,IFN)-γmRNA的表达,Western blot检测肺组织中NF-κB2蛋白的表达,ELISA检测BALF中细胞因子的表达。结果哮喘模型组中miR-27a的表达降低,miR-27a模拟物增加了miR-27a的表达,anti-miR-27a明显降低了miR-27a的表达。与OVA组相比,miR-27a模拟物处理后,小鼠的气道高反应性和BALF中炎性细胞总数、分类计数均明显降低(P<0.05),肺组织病理学变化减轻,NF-κB2 mRNA的表达无明显变化,但其蛋白表达明显减少(P<0.05),IL-4和IL-13 mRNA和蛋白表达均降低(P<0.05),IFN-γ的表达升高(P<0.05);与OVA组相比,anti-miR-27a组中NF-κB2 mRNA表达也无显著变化,其余检测指标的变化与miR-27a模拟物的作用相反。结论 miR-27a抑制OVA诱导的哮喘小鼠气道炎症可能与其靶向NF-κB2的表达进而影响NF-κB信号通路的激活相关。
Objective To investigate the effect and mechanism of miR-27a on airway inflammation in asthmatic mice. Methods Male Balb / c mice were randomly divided into normal control group, asthma model group (OVA group), asthma + miR-27a mimics group (miR-27a group), asthma + anti-miR- anti-miR-27a group) and asthma + miR-27a control group (scramble), 8 per group. Except for the normal control group, the other mice in each group were sensitized and challenged with ovalbumin (OVA) to establish asthma model. Three asthma model groups were given nasal drops 4 mg / kg corresponding drugs Intervention. The airway hyperresponsiveness and the total number of cells in the lung lavage fluid (BALF) and the number of cells were detected on the 28th day. The pathological changes of lung were detected by HE and PAS staining. The expression of miR-27a and NF- κB, interleukin (IL) -4, IL-13 and interferon (IFN) -γmRNA were detected by Western blot. The expression of NF-κB2 in lung tissue was detected by Western blot. The expression of cytokines in BALF was detected by ELISA . Results The expression of miR-27a in asthmatic model group was decreased, miR-27a mimics increased the expression of miR-27a, and anti-miR-27a significantly reduced the expression of miR-27a. Compared with the OVA group, the airway hyperresponsiveness and the total number of inflammatory cells in BALF were significantly decreased (P <0.05), the pathological changes of lung tissue were alleviated, the expression of NF- (P <0.05), while the mRNA and protein expression of IL-4 and IL-13 decreased (P <0.05) and the expression of IFN-γ increased (P <0.05) ). Compared with OVA group, there was no significant change in NF-κB2 mRNA expression in anti-miR-27a group, and the other changes were opposite to miR-27a mimics. Conclusion miR-27a inhibits the airway inflammation in OVA-induced asthmatic mice may be related to the expression of NF-κB and NF-κB signaling pathway.