FGF10干预PM诱导支气管上皮细胞COX2/PGE_(2)表达的分子机制  

Effect of fibroblast growth factor 10(FGF10)on particulate matter(PM)-induced COX2/PGE_(2) expression in bronchial epithelial cells and its mechanism

作　　者：董年 施强强 宋晨剑 刘莉[1] 陈俊杰[1] DONG Nian;SHI Qiangqiang;SONG Chenjian;LIU Li;CHEN Junjie(Department of Pulmonary and Critical Care Medicine,the First Affiliated Hospital of Wenzhou Medical University,Zhejiang Provincial Key Laboratory of Interventional Pulmonology,Wenzhou 325015,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,325015

出　　处：《浙江医学》2022年第12期1244-1249,I0003,I0004,共8页Zhejiang Medical Journal

基　　金：浙江省自然科学基金资助项目(LQ20H010002);浙江省医药卫生科技计划项目(2020RC080)。

摘　　要：目的探讨成纤维生长因子10(FGF10)对细颗粒物(PM)诱导支气管上皮细胞表达环氧合酶2/前列腺素E_(2)(COX2/PGE_(2))的作用及其分子机制。方法雄性C57BL/6小鼠给予0.5 mg/kg FGF10腹腔注射和4.0 mg/kg PM气道滴注,构建动物模型,设置空白组、FGF10组、PM组和PM+FGF10组。获取各组肺组织并进行病理分级评分,采用Western blot法检测各组肺组织中COX2的相对表达量,采用免疫荧光法检测支气管上皮中COX2的相对表达量,采用ELISA法检测肺泡灌洗液中PGE_(2)的分泌水平。先给予5 mmol/L ERK1/2抑制剂(U0126)/PI3K抑制剂(LY294002)干预,然后10μg/L FGF10干预,最后200 mg/L PM刺激支气管上皮细胞BEAS-2B,构建细胞模型,设置空白组、PM组、PM+FGF10组、PM+FGF10+LY294002组和PM+FGF10+U0126组。采用ELISA法检测各组细胞PGE_(2)分泌水平,采用Western blot法检测各组细胞COX2的表达水平。同时进行细胞计数、线粒体功能和乳酸脱氢酶分泌分析等细胞功能检测。结果动物模型中,气道滴注PM诱导C57BL/6小鼠支气管病理形态学的改变,伴随肺泡灌洗液中PGE_(2)分泌增多(P<0.05),肺组织COX2表达升高(P<0.05),支气管上皮COX2荧光强度升高(P>0.05)。FGF10干预下,PM诱导支气管炎症和COX2/PGE_(2)表达被逆转(P<0.05)。细胞模型中,PM诱导支气管上皮细胞COX2/PGE_(2)表达水平升高(P<0.05),FGF10干预下,COX2/PGE_(2)的表达均降低(P<0.05),BEAS-2B细胞计数、线粒体功能和乳酸脱氢酶分泌等细胞功能改变(P<0.05)。就分子机制而言,LY294002和U0126逆转了FGF10对PM诱导COX2/PGE_(2)表达的调控作用(P<0.05)。结论FGF10可以调控PM对支气管上皮细胞COX2/PGE_(2)的诱导表达,其分子机制涉及MAPK/ERK和PI3K/Akt信号通路。Objective To investigate the effect of fibroblast growth factor 10(FGF10)on particulate matter(PM)-induced cyclooxygenase 2/prostaglandin E_(2)(COX2/PGE_(2))expression in bronchial epithelial cells and its mechanism.Methods C57BL/6 mice were randomly divided into vehicle group,FGF10 group,PM group and PM+FGF10 group.The mice were intraperitoneally injected with 0.5 mg/kg FGF101h before intratracheally instillation of 4 mg/kg PM for 2 consecutive days.The expression of COX2 and secretion of PGE_(2)in harvested lung tissues and bronchoalveolar lavage fluid were detected.human bronchial epithelial BEAS-2B cells were pretreated with 5μmol/L U01265 or 5μmol/L LY294002 with 10μg/L FGF10 before challenged with 200 mg/L PM.The expression of COX2 from cell lysate and the secretion of PGE_(2)in culture supernatant were detected;the cell count,mitochondria quality and LDH secretion were also determined.Results Intratracheal instillation of PM resulted in airway inflammation and up-regulated expression of COX2(P<0.05),together with the upregulated secretion of PGE_(2)in bronchoalveolar lavage fluid(P<0.05).Consistently,the FGF10 exerted a similar effect on COX2/PGE_(2)in vitro along with the increasing cell count,mitochondria quality and LDH secretion(P<0.05).Both U0126 and LY294002 reversed the effect of FGF10 on COX2/PGE_(2)expression(P<0.05).Conclusion FGF10 may exert a regulatory effect on PMinduced COX2/PGE_(2)expression in bronchial epithelial cells,through MAPK/ERK and PI3K/Akt signaling pathways.

关 键 词：支气管上皮细胞 细颗粒物 成纤维细胞生长因子10 环氧合酶2 前列腺素 2 

分 类 号：X513[环境科学与工程—环境工程] R56[医药卫生—呼吸系统]