支气管哮喘小鼠肺组织miR-155的变化及对环氧合酶-2的影响

Changes of miR-155 in lung tissue of bronchial asthma mice and its effect on COX-2

目的探讨支气管哮喘(哮喘)小鼠肺组织microRNA-155(miR-155)的变化及对环氧合酶-2(COX-2)的影响。方法雌性BALB/c小鼠28只,随机分为4组:正常对照组、哮喘模型组、antagomir阴性对照组、antagomir干预组,每组7只。应用鸡卵清蛋白(OVA)腹腔注射致敏和反复超声雾化吸入复制哮喘模型,观察各组小鼠的哮喘发作表现。通过HE染色观察肺部炎症浸润情况、酶联免疫吸附试验试剂盒测定外周血清中前列腺素E2(PGE2)的浓度、蛋白质印迹(Western-blotting)检测COX-2蛋白的表达,并通过逆转录PCR(RT-PCR)检测COX-2mRNA及miR-155-5p的表达。结果哮喘模型组小鼠miR-155-5p、COX-2及PGE2的表达均较正常对照组显著增加(t=-21.420、-15.270、-9.613,P值均<0.01);antagomir干预组miR-155、COX-2和PGE2较哮喘模型组显著降低(t=-15.273、-12.421、-6.025,P值均<0.01)。肺组织miR-155-5p的表达和COX-2的含量之间呈显著正相关(r=0.977,P<0.01)。结论哮喘小鼠肺组织中miR-155-5p表达增加,可能通过调控COX-2/PGE2途径参与哮喘发病过程。

Objective To investigate the changes of microRNA-155 (miR-155) and its effect on cyclo-oxygen-ase-2 (COX-2) in lung tissue of bronchial asthma (asthma) mice. Methods Twenty-eight female BALB/c mice were randomly divided into four groups (n=7 each): control group, asthma group, antagomir negative control group and antagomir intervention group.The asthma model was induced by intraperitoneal injection of chicken egg ovalbumin (OVA) and repeated ultrasonic atomization inhalation, and observed the asthma performance of each group of mice.Hematoxylin-eosin staining was used to observe the morphological changes of lung tissue, enzyme linked immunosorbent assay kit was used to measure the concentration of prostaglandin E2 (PGE2) in peripheral serum, and Western-blotting was used to detect the expression of COX-2 protein, reverse transcription PCR was used to detecte the expression of COX-2 mRNA and miR-155-5p. Results The expressions of miR-155-5p, COX-2 and PGE2 in the asthma group was significantly higher than those in the control group (t=-21.420,-15.270,-9.613, all P<0.01). The antagomir intervention group miR-155, COX-2, and PGE2 were significantly lower than the asthma group (t=-15.273,-12.421,-6.025, all P<0.01). The expression of miR-155-5p was significantly positively correlated with COX-2 in the lung (r=0.977, P<0.01). Conclusions The expression of miR-155-5p is increased in lung tissue of asthma mice, miR-155-5p may be involved in the pathogenesis of asthma by regulating the COX-2/PGE2 pathway.