摘要
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。
Objective：To investigate the effects of JAK/STAT6 pathway in airway remodeling in the mice with bronchial asthma and influence of AG490 on airway remodeling and its mechanism.Methods：Thirty-two femal BALB/c mice were divided into four groups at random.They were control group,the asthma group,the group of AG490 intervention and the group of budesonide intervention.The mice were sensitized and challenged with ovalbumin to establish the chronical asthma model.The airway inflammation and the alteration of airway structure were observed by the means of haematoxylin-eosin（HE） and MASSON staining.The expression of p-STAT6 was examined by Western blot and immunohistochemistry analysis,while the content of cyclinD1 was detected by Western blot and RF PCR.Results：HE and MASSON’s trichrome staining showed that there were bronchial smooth muscle hypertrophy,submucosa incrassation,airway stenosis,collagen fiber increase and mass inflammatiory cells infiltration in asthmatic group.In AG490 and budesonide intervention groups,the symptoms mentioned above were much more ameliorated.The Western blot analysis showed that the expression levels of p-STAT6 and cyclin D1 in asthmatic group were higher than control group（P 〈 0.01）,but were decreased in AG490 and budesonide intervention groups（P 〈 0.01）.The result of RT-PCR showed that the level of cyclinD1 mRNA in asthmatic group was higher than control group（P 〈 0.01）,and was reduced in the AG490 and budesonide intervention groups（P 〈 0.01）.Compared with the control group,the immunohistochemistry result showed that the expression of p-STAT6 was much higher in asthmatic group（P 〈 0.01）,and between them were AG490 and budesonide intervention groups（P 〈 0.05）.Conclusions：JAK/STAT6 signal pathway involved in airway inflammation and asthma remodeling.AG490 may ameliorate the progression of airway inflammation and remodeling via regulation of JAK/STAT6 signal pathway.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2011年第3期301-307,共7页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省卫生厅"科教兴卫工程"-呼吸病学学科(实验室)开放课题资助
