烟曲霉提取物诱导气道上皮细胞铁死亡并通过分泌的三磷酸腺苷促进哮喘血管新生机制研究  

Study on the mechanism of Aspergillus fumigatus extract inducing ferroptosis in airway epithelial cells and promoting asthma angiogenesis through adenosine triphosphate secretion

作　　者：刘婷婷[1] 张文美[1] 邓静 郑帅[1] 贾立昕[1] 朴春梅[1] 杜杰[1] LIU Tingting;ZHANG Wenmei;DENG Jing;ZHENG Shuai;JIA Lixin;PIAO Chunmei;DU Jie Beijing(Anzhen Hospital,Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Diseases,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所,100029

出　　处：《心肺血管病杂志》2024年第11期1208-1215,共8页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家自然科学基金资助项目(82170409)。

摘　　要：目的:探讨小鼠哮喘早期气道炎症发展病理生理学机制,为开展气道炎症及气道重构干预提供基础。方法:C57BL/6野生型小鼠分别气道滴注0.9%氯化钠溶液或烟曲霉提取物,3周后进行肺组织切片染色,检测血管新生。烟曲霉提取物滴注第5天收集血清、肺泡灌洗液和肺叶组织,进行炎症和铁死亡的评估。分离小鼠气道上皮细胞与烟曲霉提取物共孵育,检测溶质载体家族7成员11(solute carrier family 7,member 11,SLC7A11),抗氧化酶谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4),酰基辅酶A合成酶长链家族成员4(acyl CoA synthetase long-chain family member 4,ACSL4)和转铁蛋白(transferrin,TRF)的mRNA表达水平和蛋白表达水平,检测细胞上清中ATP的浓度。分离骨髓来源的巨噬细胞给予烟曲霉提取物刺激,与ATP受体P2X7抑制剂共孵育,检测上清中趋化因子的表达。结果:烟曲霉提取物气道滴注小鼠,肺组织浸润的炎症细胞增殖增加,肺泡灌洗液中趋化因子和细胞因子增加,肺组织血管新生增加。烟曲霉提取物刺激小鼠气道上皮细胞后,气道上皮细胞铁死亡抑制分子SLC7A11,GPX4表达降低,ACSL4表达升高,TRF表达升高,释放ATP增加。烟曲霉提取物孵育巨噬细胞,趋化因子浓度升高,给于ATP受体P2X7抑制剂后,趋化因子浓度降低。结论:烟曲霉提取物促进气道上皮细胞发生铁死亡,气道上皮细胞分泌的ATP促进炎症早期巨噬细胞分泌趋化因子,促进小鼠气道肺组织血管新生。Objective:To explore the pathological and physiological mechanisms of early development of airway inflammation in mice with asthma,and to provide a basis for intervention in airway inflammation and airway remodeling.Methods:C57BL/6 wild-type mice were intratracheal instillation saline or Aspergillus fumigatus extract,and after 3 weeks,lung tissue sections were stained to detect angiogenesis.On the 5th day of intratracheal instillation of Aspergillus fumigatus extract,serum,bronchoalveolar lavage fluid,and lung tissue were collected for evaluation of inflammation and ferroptosis.Mouse airway epithelial cells were isolated and co-cultured with Aspergillus fumigatus extract.The mRNA and protein expression levels of SLC7A11(solute carrier family 7,Member 11),GPX4(glutathione peroxidase 4),ACSL4(acyl CoA synthetase long-chain family member 4),and TRF(transferrin)were detected,and the concentration of ATP in the cell supernatant was measured.Bone marrow macrophages were stimulated with Aspergillus fumigatus extract and co-incubated with ATP receptor P2X7 inhibitor to detect the expression of chemokines in the supernatant.Results:The airway intratracheal instillation of Aspergillus fumigatus extract into mice resulted in increased inflammatory cells infiltrating lung tissue,increased chemokines and cytokines in alveolar lavage fluid,and increased angiogenesis in lung tissue.After stimulation of mouse airway epithelial cells with Aspergillus fumigatus extract,the expression of ferroptosis inhibitor molecule SLC7A11 and GPX4 in airway epithelial cells decreased,ACSL4 expression increased,transferrin expression increased,and ATP release increased.After incubating bone marrow derived macrophages with Aspergillus fumigatus extract,the concentration of chemokines increased.After administration of ATP receptor P2X7 inhibitor,the concentration of chemokines decreased.Conclusion:Aspergillus fumigatus extract promotes ferroptosis in airway epithelial cells,and the ATP secreted by airway epithelial cells promotes the secretion of

关 键 词：气道上皮细胞 三磷酸腺苷 铁死亡 巨噬细胞 趋化因子 

分 类 号：R54[医药卫生—心血管疾病]