Xiong Jing, Zhao Wen-Qu, Huang Guo-Hua, Yao Li-Hong, Dong Hang-Ming, Yu Chang-Hui, Zhao Hai-Jin, Cai Shao-Xi
Laboratory of Chronic Airway Diseases, Department of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Oct 20;37(10):1301-1307. doi: 10.3969/j.issn.1673-4254.2017.10.04.
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.
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 16HBE was transfected with lentiviral vector carrying short hairpin RNA targeting RAGE (shRNA?RAGE) and subsequently challenged with a TDI?human serum albumin (TDI-HSA) 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.
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 16HBE 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?HSA-induced 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).
RAGE signaling induces MUC5AC expression via extracellular signal-regulated kinase pathway to promote mucus overproduction in mice with TDI-induced asthma.
探讨晚期糖基化终末产物受体（RAGE）在调节甲苯二异氰酸酯（TDI）诱导的哮喘小鼠模型中黏蛋白5AC（MUC5AC）表达及黏液分泌中的作用。
将BALB/c小鼠随机分为对照组、溶剂（AOO）组、TDI诱导的哮喘组和RAGE抑制剂（FPS-ZM1）组。采用过碘酸雪夫（PAS）染色、蛋白质印迹法和免疫组织化学法分析小鼠气道黏液分泌及MUC5AC表达的变化，并用蛋白质印迹法检测p-ERK的表达。用携带靶向RAGE的短发夹RNA（shRNA-RAGE）的慢病毒载体转染体外培养的人支气管上皮细胞系16HBE，随后用TDI-人血清白蛋白（TDI-HSA）偶联物刺激，用逆转录聚合酶链反应（RT-PCR）检测细胞内MUC5AC mRNA表达的变化；用蛋白质印迹法检测细胞中ERK和p-ERK的蛋白表达。还分析了ERK抑制剂U0126预处理对细胞中MUC5AC mRNA表达的影响。
与对照小鼠相比，TDI诱导的哮喘小鼠气道PAS阳性率显著升高，MUC5AC和p-ERK表达增加（P<0.05）。FPS-ZM1处理显著降低了哮喘小鼠气道的PAS阳性率，降低了MUC5AC和p-ERK的表达（P<0.05）。16HBE细胞暴露于TDI-HSA后，MUC5AC mRNA表达和p-ERK蛋白表达显著增加（P<0.05），而RAGE基因敲低明显抑制了TDI-HSA诱导的p-ERK和MUC5AC mRNA上调（P<0.05）。ERK抑制剂U0126处理也降低了TDI-HSA诱导的细胞中MUC5AC mRNA上调（P<0.05）。
RAGE信号通路通过细胞外信号调节激酶途径诱导MUC5AC表达，促进TDI诱导的哮喘小鼠黏液过度分泌。
