自噬流在细颗粒物诱导的气道炎症中的变化及成纤维细胞生长因子10调控自噬流的抗炎效应  

Change of autophagic flux in particulate matter-induced airway inflammation and anti-inflammatory effect of fibroblast growth factor 10 related to regulating autophagic flux

作　　者：董年 王强[1] 施强强 刘玲静 陈俊杰[1] DONG Nian;WANG Qiang;SHI Qiang-qiang;LIU Ling-jing;CHEN Jun-jie(Key Laboratory of Interventional Pulmonology of Zhejiang Province,Department of Respiratory and Critical Care Medicine,The First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第一医院呼吸与危重症医学科,浙江省介入肺脏病学重点实验室,浙江温州325000

出　　处：《中国病理生理杂志》2021年第10期1815-1821,共7页Chinese Journal of Pathophysiology

基　　金：浙江省自然科学基金资助项目(No.LQ20H010002);浙江省医药卫生科技计划项目(No.2020361781,No.2022480227);温州市科技局课题(No.Y20180125,No.Y20190522)。

摘　　要：目的:探讨自噬流在细颗粒物(PM)诱导气道炎症中的变化及成纤维细胞生长因子10(FGF10)是否通过调控自噬流发挥抗炎效应。方法:选取雄性C57BL/6小鼠进行随机分组,预先1 h给予5 mg/kg FGF10气道滴注,然后予以4 mg/kg PM气道滴注,连续气道滴注2 d,构建动物模型,具体分组:空白对照(vehicle)组、FGF10组、PM组和PM+FGF10组。获取肺组织进行病理分级评分,获取支气管肺泡灌洗液(BALF)以ELISA检测炎症介质白细胞介素6(IL-6)、IL-8、前列腺素E2(PGE2)和肿瘤坏死因子α(TNF-α)的分泌,获取肺组织蛋白以Western blot检测自噬相关蛋白LC3和p62的表达。预先给予4 mmol/L 3-甲基腺嘌呤(3-MA)或10μg/L FGF10干预1 h,200 mg/L的PM刺激人正常气道上皮BEAS-2B细胞1 h,Western blot检测NF-κB通路p65和IκBα磷酸化水平。预先给予4mmol/L 3-MA或10μg/L FGF10干预1 h,200 mg/L的PM刺激BEAS-2B细胞24 h,ELISA检测炎症介质IL-6、IL-8、PGE2和TNF-α的分泌,Western blot检测自噬相关蛋白LC3和p62的表达。结果:动物模型中PM短时暴露可以诱导C57BL/6小鼠气道炎症的改变,伴随BALF中炎症介质IL-6、IL-8、PGE2和TNF-α升高,肺组织发生自噬流异常;FGF10干预可以逆转PM诱导的气道炎症和炎症介质分泌,伴随着自噬流的改变。细胞模型中PM可以诱导BEAS-2B细胞炎症介质IL-6、IL-8、PGE2和TNF-α分泌,3-MA干预或FGF10干预可以逆转炎症介质的分泌,3-MA干预可以逆转NF-κB通路的改变,FGF10干预可以逆转自噬流的改变。结论:FGF10在PM诱导的气道炎症中发挥抗炎效应,其中分子机制涉及自噬和NF-κB信号通路。AIM:To investigate the change of autophagic flux in particulate matter(PM)-induced airway inflammation,and to explore whether fibroblast growth factor 10(FGF10)exerts its anti-inflammatory effect via regulating autophagic flux.METHODS:In vivo,the C57 BL/6 mice were randomly grouped,with intratracheal instillation of 5 mg/kg FGF10 1 h before 4 mg/kg PM for 2 consecutive days. The harvested lung tissues were scored by pathological grading.In vitro,normal human airway epithelial BEAS-2 B cells were pretreated with 4 mmol/L 3-methyladenine(3-MA)or 10 μg/L FGF10 before challenged with 200 mg/L PM. The levels of inflammatory mediators interleukin-6(IL-6),IL-8,prostaglandin E2(PGE2)and tumor necrosis factor-α(TNF-α)in culture supernatant and bronchoalveolar lavage fluid(BALF)were quantified by ELISA. The levels of autophagy-related proteins(LC-3 and p62)and NF-κB signaling pathway-related proteins(p65,p-p65,IκBα and p-IκBα)from cell lysate and lung tissue were determined by Western blot.RESULTS:In vivo,short-term PM exposure resulted in airway inflammation characterized by infiltration of inflammatory cells and upregulated secretion of inflammatory mediators IL-6,IL-8,PGE2 and TNF-α,as well as the up-regulated expression of LC3-II and p62,while FGF10 exerted a reverse effect. Consistently,FGF10 exerted a similar anti-inflammatory effect in vitro along with the reverse of autophagic flux. Moreover,3-MA reversed the phosphorylation of p65 and IκBα.CONCLUSION:FGF10 exerts an anti-inflammatory effect on PM-induced airway inflammation through autophagy and NF-κB signaling pathway.

关 键 词：细颗粒物 自噬流 成纤维细胞生长因子10 气道炎症 NF-ΚB信号通路 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R363.2[医药卫生—基础医学]