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目的:探讨Janus激酶/信号转导与活化蛋白6(JAK/STAT6)在支气管哮喘小鼠气道重塑中作用及AG490影响气道重塑的可能机制。方法:将32只雌性BALB/c小鼠随机分为4组,正常对照组(A)、慢性哮喘模型组(B)、AG490干预组(C)和布地奈德干预组(D),每组8只。应用鸡卵清蛋白致敏和激发建立慢性哮喘气道重塑模型,HE和MASSON染色观察各组小鼠气道炎症和气道结构的改变,免疫印迹法测定各组小鼠肺组织中p-STAT6和细胞周期蛋白D1(cyclinD1)的表达水平,免疫组化法检测各组小鼠肺组织中p-STAT6的表达情况,用半定量逆转录-聚合酶链反应(RT-PCR)法测定各组小鼠肺组织中cyclinD1mRNA的数量变化。结果:HE和MASSON染色结果提示哮喘组黏膜下层和平滑肌层增厚,气道管腔狭窄,胶原纤维增生,大量炎症细胞浸润,AG490和布地奈德干预组上述改变较哮喘组为轻;免疫印迹结果提示哮喘组p-STAT6和cyclinD1蛋白的水平均较空白对照组显著增高(P<0.01),AG490和布地奈德干预组p-STAT6和cyclinD1蛋白的水平较哮喘组降低;RT-PCR结果显示哮喘组cyclinD1 mRNA含量较正常对照组明显增多(P<0.01),在AG490和布地奈德干预组有所降低(P<0.01)。免疫组化结果显示与正常对照组比较,p-STAT6的表达在哮喘模型组的气道上皮细胞中显著增多(P<0.01),而在AG490和布地奈德干预组的表达介于前两者之间(P<0.05)。结论:JAK/STAT6信号通路活化可能是哮喘气道炎症和气道重塑的病理机制之一,AG490可能通过抑制JAK/STAT6信号通路的活化,进而减轻哮喘小鼠气道炎症和气道重塑。
AIM: To investigate the role of Janus kinase / signal transducer and activator of transcription 6 (JAK / STAT6) in airway remodeling in asthmatic mice and the possible mechanism of AG490 in airway remodeling. Methods: 32 female BALB / c mice were randomly divided into 4 groups: normal control group (A), chronic asthma model group (B), AG490 intervention group (C) and budesonide intervention group 8 only The model of airway remodeling was established by sensitizing and stimulating chicken ovalbumin. The airway inflammation and airway structure of mice were observed by HE and MASSON staining. The expression of p-STAT6 and The expression of cyclinD1 was detected by immunohistochemistry. The expression of p-STAT6 in lung tissue of each group was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) Changes in the number of cyclinD1 mRNA in murine lung tissue. Results: The results of HE and MASSON staining showed that the thickening of the submucosa and smooth muscle layer, the narrowing of airway lumen, the proliferation of collagen fibers and the infiltration of a large number of inflammatory cells in asthma group and the mild changes in the AG490 and budesonide groups were less than those in the asthma group. The results showed that the levels of p-STAT6 and cyclinD1 protein in asthma group were significantly higher than those in blank control group (P <0.01), and the levels of p-STAT6 and cyclinD1 in AG490 and budesonide groups were lower than those in asthma group The level of cyclinD1 mRNA in asthma group was significantly higher than that in normal control group (P <0.01), but decreased in AG490 and budesonide groups (P <0.01). The results of immunohistochemistry showed that the expression of p-STAT6 was significantly increased in the airway epithelial cells of the asthma model group compared with the normal control group (P <0.01), while the expression of p-STAT6 was higher in the AG490 and budesonide intervention groups (P <0.05). Conclusion: Activation of JAK / STAT6 signaling pathway may be one of the pathological mechanisms of airway inflammation and airway remodeling in asthma. AG490 may reduce airway inflammation and airway remodeling in asthmatic mice by inhibiting the activation of JAK / STAT6 signaling pathway.