香烟烟雾联合PolyI:C诱导COPD模型小鼠气道上皮屏障损伤的机制  

Mechanism of airway epithelial barrier injury in a mouse model of COPD induced by cigarette smoke exposure combined with Poly I:C

作　　者：谢媛 梅晓峰[3] 陶柳颖 江宇航 李建生[1,2,3] 赵鹏 XIE Yuan;MEI Xiaofeng;TAO Liuying;JIANG Yuhang;LI Jiansheng;ZHAO Peng(Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province&Educa-tion Ministry of China,Henan Key Laboratory of Chinese Medicine for Respiratory Disease,Henan University of Chinese Medicine,Zhengzhou 450046,China;Academy of Chinese Medical Sciences,Henan University of Chinese Medicine,Zhengzhou 450046,China;Department of Respiratory Diseases,The First Affiliated Hospital of Henan University of Chi-nese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学呼吸疾病中医药防治省部共建协同创新中心,河南省中医药防治呼吸病重点实验室,河南郑州450046 [2]河南中医药大学中医药科学院,河南郑州450046 [3]河南中医药大学第一附属医院,河南郑州450046

出　　处：《中国病理生理杂志》2024年第7期1222-1229,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81973822);河南省科技研发计划联合基金项目(No.22301420070);郑州市科技协同创新专项(No.2023XTCX045)。

摘　　要：目的:用香烟烟雾(CS)暴露联合聚肌胞苷酸(Poly I:C)滴鼻建立慢性阻塞性肺疾病(COPD)小鼠模型,探讨COPD气道上皮屏障损伤的机制。方法:(1)将96只雄性BALB/c小鼠随机分为对照组、CS组、Poly I:C组和CS+Poly I:C组,每组24只。第1~8周造模,每4周检测肺功能,于第4、8、16和24周末取材,每组取6只。测定每分钟通气量(MV)、气道狭窄指数(Penh)、肺泡平均截距(MLI)和支气管管壁厚度(BWT)的变化,检测白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、闭锁小带蛋白1(ZO-1)和上皮钙黏素(E-Cad)水平。(2)用CS提取物(CSE)联合Poly I:C刺激人支气管上皮BEAS-2B细胞24 h,检测ZO-1、闭合蛋白(Occ)、磷酸化表皮生长因子受体(EGFR)、磷酸化P38和磷酸化细胞外信号调节激酶(ERK)1/2蛋白水平。结果:(1)与对照组相比,第8周CS组和CS+Poly I:C组小鼠肺组织出现大量炎症细胞浸润、肺泡腔扩张、肺泡壁断裂融合、气管壁增厚等病理变化,Penh、BWT、MLI、IL-1β和TNF-α显著升高(P<0.05或P<0.01),MV、ZO-1和E-Cad显著降低(P<0.05或P<0.01);第24周,CS+Poly I:C组以上病理变化仍较稳定存在。(2)与对照组相比,CSE联合Poly I:C显著诱导BEAS-2B细胞ZO-1和Occ蛋白表达降低(P<0.05或P<0.01),EGFR、P38和ERK1/2磷酸化水平升高(P<0.01);且CSE联合Poly I:C组显著优于CSE组和Poly I:C组。结论:Poly I:C可以促进CS诱导的COPD模型小鼠病理改变和气道上皮屏障损伤,其机制可能与激活EGFR/ERK/P38信号通路有关。AIM:To establish a mouse model of chronic obstructive pulmonary disease(COPD)induced by cigarette smoke(CS)exposure combined with polyinosinic-polycytidylic acid(Poly I:C)nasal drip,and to investigate the mechanism of airway epithelial barrier injury in COPD.METHODS:(1)Ninety-six male BALB/c mice were randomly divided into control group,CS group,Poly I:C group,and CS+Poly I:C group(n=24).The model was established from week 1 to week 8,with pulmonary function tested every 4 weeks.Six mice from each group were sacrificed at the end of weeks 4,8,16,and 24.Changes in minute volume(MV),enhanced pause(Penh),mean linear intercept(MLI)and bronchial wall thickness(BWT)were observed.The protein levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),zonula occludens-1(ZO-1)and E-cadherin(E-Cad)in the lung were detected.(2)Human bronchial epithelial BEAS-2B cells were stimulated with CS extract(CSE)combined with Poly I:C for 24 h,and then the protein levels of occludin(Occ),ZO-1,and phosphorylated epidermal growth factor receptor(EGFR),P38 and extracellular signal-regulated kinase(ERK)1/2 were analyzed.RESULTS:(1)Compared with control group,at the 8th week,the mice in CS and CS+Poly I:C groups showed typical pathological changes in lung tissues,including significant inflammatory cell infiltration,alveolar cavity expansion,alveolar wall rupture and fusion,and airway wall thickening.The Penh,BWT,MLI,and lung IL-1βand TNF-αlevels were significantly increased(P<0.05 or P<0.01),while MV and lung ZO-1 and E-Cad levels were remarkably decreased(P<0.05 or P<0.01).By the 24th week,these pathological changes remained relatively stable in CS+Poly I:C group.(2)Compared with control group,CSE and its combination with Poly I:C dramatically induced a reduction in ZO-1 and Occ protein expression in BEAS-2B cells(P<0.05 or P<0.01),and increased the levels of phosphorylated EGFR,P38 and ERK1/2(P<0.01).The effects in CSE combined with Poly I:C group were considerably superior to those in CSE or Poly I:C group alone.CONCLUSION:Pol

关 键 词：慢性阻塞性肺疾病 香烟烟雾 聚肌胞苷酸 气道上皮屏障 

分 类 号：R563[医药卫生—呼吸系统] R363[医药卫生—内科学] R122[医药卫生—临床医学]