基于MAPK/ERK信号通路探讨孟鲁司特通过自噬改善哮喘模型小鼠肺损伤的机制  

Mechanism of montelukast in improving lung injury in asthmatic mice through autophagy based on MAPK/ERK signaling pathway

作　　者：林妍妍 裴剑 夏彦东 LIN Yanyan;PEI Jian;XIA Yandong(Department of Pharmacy,the Third Hospital of Tangshan,Tangshan,Hebei 063000,China;Department of Neurosurgery,Tangshan Gongren Hospital,Tangshan,Hebei 063000,China;Clinical Laboratory Department,Kailuan General Hospital,Tangshan,Hebei 063000,China)

机构地区：[1]河北省唐山市第三医院药剂科,河北唐山063000 [2]河北省唐山市工人医院神经外科,河北唐山063000 [3]开滦总医院检验科,河北唐山063000

出　　处：《检验医学与临床》2024年第20期3033-3037,3043,共6页Laboratory Medicine and Clinic

基　　金：河北省中医药管理局中医药科研计划项目(2020420)。

摘　　要：目的探究孟鲁司特是否可改善哮喘小鼠肺损伤,并基于丝裂原活化蛋白激酶(MAPK)/细胞外信号调节蛋白激酶(ERK)信号通路介导的自噬探究其改善肺损伤的机制。方法选取60只无特定病原体(SPF)级BALB/c小鼠:对照组、哮喘组、孟鲁司特组及司美替尼(Selumetinib)组,每组15只。肺功能仪检测小鼠第0.15秒用力呼气容积(FEV_(0.15))、最高呼气流速(PEF)。苏木素-伊红(HE)染色检测小鼠肺组织病理学形态。Masson染色检测肺组织平滑肌增生与胶原沉积水平。Western blotting检测肺组织中MAPK、ERK、Beclin1及微管相关蛋白轻链3(LC3Ⅱ)蛋白水平。免疫组织化学检测肺组织中MAPK、ERK蛋白水平。结果与对照组比较,哮喘组小鼠FEV_(0.15)、PEF均降低,差异均有统计学意义(P<0.05);与哮喘组比较,孟鲁司特组FEV_(0.15)、PEF均增加,差异均有统计学意义(P<0.05)。HE染色结果显示,哮喘组小鼠可见支气管壁增厚、炎性浸润显著等病理学形态损伤;孟鲁司特组可见肺组织病理学形态损伤改善。Masson染色结果显示,哮喘组小鼠支气管平滑肌增厚,且其周围可见胶原大量沉积;孟鲁司特组小鼠支气管平滑肌增生减轻,周围胶原沉积减少。Western blooting检测结果显示,与对照组比较,哮喘组小鼠肺组织中MAPK、ERK、Beclin1、LC3Ⅱ表达水平增加,差异均有统计学意义(P<0.05);与哮喘组比较,孟鲁司特组小鼠肺组织中MAPK、ERK、Beclin1、LC3Ⅱ表达水平降低,差异均有统计学意义(P<0.05)。与对照组比较,哮喘组小鼠FEV_(0.15)、PEF降低,MAPK、ERK、Beclin1、LC3Ⅱ蛋白表达水平均增加,差异均有统计学意义(P<0.05)。与哮喘组比较,Selumetinib组FEV_(0.15)、PEF均增加,MAPK、ERK、Beclin1、LC3Ⅱ蛋白表达水平均降低,差异均有统计学意义(P<0.05)。结论孟鲁司特可改善哮喘小鼠肺损伤,其机制可能与MAPK/ERK信号通路介导的自噬相关。Objective To investigate whether montelukast can improve lung injury in asthmatic mice,and to explore its possible protective mechanism based on autophagy mediated by mitogen-activated protein kinase(MAPK)/extracellular signal-regulated protein kinase(ERK)signaling pathway.Methods Sixty specific pathogen free(SPF)BALB/c mice were randomly divided into control group,asthma group,montelukast group and Selumetinib group,with 15 mice in each group.Forced expiratory volume in 0.15 second(FEV_(0.15))and peak expiratory flow(PEF)were measured by pulmonary function analyzer.Hematoxylin-eosin(HE)staining was used to detect the pathological morphology of lung tissue.Masson staining was used to detect the proliferation of smooth muscle and collagen deposition in lung tissue.The protein lev els of MAPK,ERK,Beclin1 and microtubule-associated protein light chain 3Ⅱ(LC3Ⅱ)in lung tissue were detected by Western blotting.The protein levels of MAPK and ERK in lung tissue were detected by immunohistochemistry.Results Compa red with the control group,FEV_(0.15) and PEF in the asthma group were decreased,and the differences were statistically significant(P<0.05).Compared with the asthma group,FEV_(0.15) and PEF in the montelukast group increased,and the differences were statistically significant(P<0.05).The results of HE staining showed that the mice in the asthma group had pathological morphological damage such as bronchial wall thickening and inflammatory infiltration.The pathological changes of lung tissue were improved in the montelukast group.Masson staining showed that the bronchial smooth muscle of the asthma group was thickened,and a large amount of collagen was deposited around it.In montelukast group,bronchial smooth muscle hyperplasia and collagen deposition were reduced.Western blotting showed that compared with the control group,the asthma group had significant increases in the protein expressions of MAPK,ERK,Beclin1 and LC3Ⅱin lung tissue(P<0.05).Compared with the asthma group,the montelukast group had significant

关 键 词：哮喘 肺损伤 孟鲁司特 MAPK/ERK信号通路 自噬 

分 类 号：R392.8[医药卫生—免疫学] R563[医药卫生—基础医学]