晚期糖基化终末产物受体促进甲苯二异氰酸脂哮喘小鼠MUC5AC表达及气道黏液高分泌  

Receptor for advanced glycation end products upregulates MUC5AC expression and promotes mucus overproduction in mice with toluene diisocyanate-induced asthma

作　　者：熊婧[1] 赵文驱 黄国华[1] 姚利红[1] 董航明[1] 余常辉[1] 赵海金[1] 蔡绍曦[1] 

机构地区：[1]南方医科大学南方医院呼吸与危重症医学科//慢性气道疾病实验室,广东广州510515

出　　处：《南方医科大学学报》2017年第10期1301-1307,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(81670026);国家重点研发计划精准医学研究发展计划(2016YFC0905800);广东省自然科学基金(2015A030313236);广东省科技计划项目(2016A020215117)~~

摘　　要：目的探讨晚期糖基化终末产物受体(RAGE)信号对甲苯二异氰酸脂(TDI)哮喘小鼠黏蛋白MUC5AC表达及气道黏液分泌的影响。方法在体内水平上建立TDI小鼠哮喘模型,实验分4组:对照组、TDI溶剂(AOO)致敏激发组、TDI致敏激发组、RAGE抑制剂处理组。采用糖原染色评估各组间气道黏液分泌情况,Western blotting及免疫组化法检测MUC5AC表达水平。同时通过Western blotting检测各组间细胞外调节蛋白激酶(ERK)通路磷酸化水平。在体外水平配制TDI-人血清白蛋白(TDIHSA)复合物,刺激人支气管上皮细胞(16HBE)及经慢病毒转染空载体和sh RNA-RAGE载体的人支气管上皮细胞,检测各组MUC5AC及ERK通路分子表达情况,加入ERK抑制剂U0126后,进一步评估MUC5AC mRNA表达情况。结果与对照组相比,TDI哮喘组小鼠糖原染色阳性率及MUC5AC表达升高(P<0.05),p-ERK表达增多(P<0.05),RAGE抑制剂处理组糖原染色阳性率及MUC5AC表达较TDI组减少(P<0.05)。同时,p-ERK表达下降(P<0.05)。体外水平,与对照组相比,TDI-HSA处理组MUC5AC mRNA及p-ERK表达增多(P<0.05),sh RNA-RAGE组MUC5AC mRNA及p-ERK表达下调(P<0.05),ERK抑制剂预处理组,MUC5AC mRNA表达下调(P<0.05)。结论 TDI小鼠哮喘模型下RAGE可能通过激活ERK通路促进黏蛋白MUC5AC的表达及黏液高分泌的发生。Objective To explore the role of the receptor for advanced glycation end products（RAGE） in regulating the expression of MUC5AC and mucus production in a mouse model of toluene diisocyanate（TDI）-induced asthma.Methods BALB/c mice were randomly divided into control group,vehicle（AOO） group,TDI-induced asthma group and RAGE inhibitor（FPS-ZM1） group.PAS staining,Western blotting,and immunohistochemistry were used to analyze the changes in mucus production and MUC5AC expression in the airway of the mice,and the expression of p-ERK was detected with Western blotting.In vitro cultured human bronchial epithelial cell line 16 HBE was transfected with lentiviral vector carrying short hairpin RNA targeting RAGE（sh RNA-RAGE） and subsequently challenged with a TDI-human serum albumin（TDIHSA） conjugate,and the changes in cellular MUC5AC mRNA expression as detected using RT-PCR;the protein expressions of ERK and p-ERK in the cells were examined with Western blotting.The effect of ERK inhibitor U0126 pretreatment on MUC5AC mRNA expression was also analyzed in the cells.Results Compared with the control mice,TDI-induced asthmatic mice showed significantly higher rates of PAS positivity and increased MUC5AC and p-ERK expressions in the airway（P〈0.05）.Treatment with FPS-ZM1 significantly decreased PAS positivity and lowered MUC5AC and p-ERK expressions in the airway of the asthmatic mice（P〈0.05）.Exposure of 16 HBE cells to TDI-HSA caused a significant increase in MUC5AC mRNA expression and p-ERK protein expression（P〈0.05）,while RAGE knockdown obviously suppressed TDI-HSAinduced upregulation of p-ERK and MUC5AC mRNA（P〈0.05）.Treatment with the ERK inhibitor U0126 also lowered TDI-HSA-induced up-regulation of MUC5AC mRNA in the cells（P〈0.05）.Conclusion RAGE signaling induces MUC5AC expression via extracellular signalregulated kinase pathway to promote mucus overproduction in mice with TDI-induced asthma.

关 键 词：甲苯二异氰酸酯哮喘 晚期糖基化终末产物受体 MUC5AC P-ERK 

分 类 号：R562.25[医药卫生—呼吸系统]