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CD39/ATP轴调控哮喘小鼠肺泡巨噬细胞炎症小体活化的研究

CD39/ATP axis regulates inflammasome activation of alveolar macrophages in asthmatic mouse

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【作者】 张方施毅宋勇辛晓峰方丽萍朱美英沈华浩

【Author】 ZHANG Fang;SHI Yi;SONG Yong;XIN Xiao-feng;FANG Li-ping;ZHU Mei-ying;SHEN Hua-hao;Department of Respiration Diseases,Nanjing General Hospital of Nanjing Military Command;

【机构】 南京军区南京总医院呼吸科浙江大学医学院附属第二医院呼吸内科

【摘要】 目的 探讨CD39/ATP轴与哮喘肺泡巨噬细胞炎症小体活化之间的关系。方法 采用OVA致敏和激发模式建立Th2优势哮喘小鼠模型;采用LPS联合OVA致敏和OVA激发模式建立Thl7优势哮喘小鼠模型;将CD39抗体经鼻滴入至Th2优势哮喘小鼠,建立CD39阻断的哮喘小鼠模型。实验设立正常对照组、Th2优势哮喘组、Th17优势哮喘组以及CD39阻断组,每组小鼠各10只,共40只。分离各组小鼠肺泡巨噬细胞,采用流式细胞仪检测各组肺泡巨噬细胞CD39的表达,Western Blot检测巨噬细胞炎症小体分子NLRP3和caspase-1的表达,ELISA方法 检测肺泡灌洗液中ATP、IL-1β的含量。结果 Th17优势哮喘组以及CD39阻断组小鼠肺泡巨噬细胞CD39表达明显下调,与之对应的是:炎症小体中NLRP3和caspase-1的蛋白表达上调和肺泡灌洗液中ATP、IL-1β含量显著增加。结论 哮喘小鼠肺泡巨噬细胞CD39表达与炎症小体活化之间存在着相关性,推测CD39/ATP轴在调控哮喘小鼠肺泡巨噬细胞炎症小体活化之中发挥重要作用。

【Abstract】 Objective To investigate the association between CD39/ATP axis and inflammasome activation of alveolar macrophages in asthmatic mouse.Methods The Th2 dominant mouse model of asthma were establish by the OVA-sensitized and provocated protocol;Also,the Th17 dominant mouse model of asthma were establish by using LPS plus OVA-sensitized and OVA-provocated protocol;And the blocking CD39 mouse model of asthma were established by using,CD39 antibody instilled intranasally into Th2 dominant mouse model of asthma.The experiment include the control group,Th2 dominant asthma group,Th17 dominant asthma group and CD39-blocking group.The mouse number in each group was 10,and40 in total.Alveolar macrophages were isolated from the mouse in each group.The expression of CD39 in alveolar macrophages were evaluated by flow cytometry,and the NLRP3,ASC and caspase-1 expression were analyzed by Western Blot,and the levels of ATP,IL-1β in bronchoalveolar fluid were detected by ELISA.Results CD39 expression on alveolar macrophages in the mouse of Th17 dominant asthma group and CD39-blocking group,were significantly reduced;Correspondingly,the expression of NLRP3,ASC and caspase-1 protein in inflammasome and the levels of ATP,IL-ip in bronchoalveolar fluid were significantly increased.Conclusion The CD39 expression on alveolar macrophages has an association with inflammasome activation in asthmatic mouse,suggesting CD39/ATP axis in the regulation of inflammasome activation in the alveolar macrophage of asthmatic mice.

【关键词】 CD39ATP巨噬细胞炎症小体
【Key words】 CD39ATPMacrophagesInflammasome
【基金】 国家自然科学基金(81270079)
  • 【文献出处】 老年医学与保健 ,Geriatrics & Health Care , 编辑部邮箱 ,2014年06期
  • 【分类号】R562.25
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