JNK12-AP1信号转导通路对PM2.5所致气道黏液高分泌的介导作用.docVIP

基金项目：国家自然科学基金1作者单位：重庆医科大学附属第二医院呼吸科,E-mail:zywenjianli@163.com1*通讯作者：周向东, 基金项目：国家自然科学基金 1作者单位：重庆医科大学附属第二医院呼吸科,E-mail:zywenjianli@163.com 1*通讯作者：周向东, E-mail:zxd999@263.net 泌的介导作用 温建立1 李琪1 周向东1* （重庆医科大学附属第二医院呼吸内科，重庆 400010;） 摘要：目的 研究直径≤2.5μm 空气中可吸入颗粒（PM2.5）所致气道黏液高分泌的信号转导机制。方法 PM2.5刺激大鼠建立气道黏液高分泌模型，Wistar大鼠随机分为对照组、PM2.5组、PM2.5+SP600125组。ELISA检测各实验组中粘蛋白（MUC）5AC在蛋白水平的表达量，Real time-PCR检测MUC5AC 在mRNA水平的表达量，Western blotting检测JNK 3个亚型（p-JNK1、p-JNK2、p-JNK3）的蛋白含量，EMSA检测该信号通路下游激活蛋白1（AP-1）与DNA的结合活性。结果 PM2.5刺激组与空白对照组相比， 前者MUC5AC在mRNA水平和蛋白水平的表达量均显著高于后者（P＜0.01），通过Western Blotting法测得p-JNK1、p-JNK2的含量也明显高于对照组（P＜0.01），p-JNK3的蛋白含量则无明显变化，EMSA结果提示：PM2.5刺激大鼠后，测得激活蛋白1结合活性明显升高（P＜0.01），给予JNK特异性抑制剂SP600125后，测得激活蛋白1结合活性明显降低（P＜0.05）。与PM2.5刺激组比较，PM2.5+SP600125组的MUC5AC在mRNA水平和蛋白水平的表达明显降低（P＜0.05）。结论 PM2.5可以通过JNK1/2-AP-1信号传导通路调节气道黏液高分泌。 关键词：PM2.5; c-Jun N端激酶; 气道黏液高分泌; 信号传导 中图分类号： 文献标识码： PM2.5 induce mucus secrection via JNK1/2-AP-1 pathway in vitro WEN Jian Li1, LI Qi1, ZHOU Xiang Dong1* Abstract：Objective To investigate the mechanism of PM2.5 on airway mucous hypersecretion. Methods Stimulated rats bronchial epithelial cell with PM2.5 to create airway mucous hypersecrection model in vitro , Wistar rats were divided into 3 groups: control group, PM2.5 group and PM2.5+SP600125 group. The mRNA and protein of MUC5AC were measured by real-time PCR and ELISA, respectively. The subtype of p-JNK (p-JNK1, p-JNK2, p-JNK3 ) were measured by western blotting. Electrophoretic mobitity shift assay (EMSA) was used to detect the DNA-binding activities of activator protein-1. Results As compared with the control group, the mRNA and protein of MUC5AC and p-JNK1, p-JNK2, in PM2.5 group were obviously higher than those of the control group (all P＜0.01). but did not affect the phosphorylation of JNK3, The DNA-binding activities of activator protein-1 was up-regulated in PM2.5 group by EMSA, The DNA-binding activities of activator protein-1 was down-regulated in PM2.5+SP60