离体热损伤成纤维细胞所涉及的细胞信号通路  

SIGNAL PATHWAY INVOLVED IN HEAT-STRESSED FIBROBLASTS

作　　者：程飚[1] 付小兵[1] 盛志勇[1] 张萍[2] 孟晋红[2] 

机构地区：[1]解放军第304医院全军烧伤研究所,北京100037 [2]第四军医大学基础部全军神经科学研究所

出　　处：《解放军医学杂志》2003年第7期590-591,I004,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家重点基础研究规划 (编号G1 9990 542 0 4 );国家自然科学基金重点项目 (编号 30 2 30 370 );国家杰出青年科学基金 (编号 3952 50 2 4 );国家自然科学基金 (编号 30 1 70 966);博士后基金 (编号中博基 2 0 0 1 1 4 )资助课题

摘　　要：利用已建立的成纤维细胞热损伤诱导凋亡模型观察离体培养的成纤维细胞热烫伤后信号通路的活化情况。首先将原代培养的人成纤维细胞经 5～ 6传代后 ,45℃水中孵育 10min,造成单纯热损伤刺激 ,加入 5 %小牛血清继续培养 ,分别在伤后 0、3 0、60和180min 4个时相点冻存细胞 ,运用Westernblot进行MAPKs信号通路磷酸化蛋白印迹检测 ,同时采用荧光显微镜检测热损伤成纤维细胞caspase3蛋白的表达情况。结果显示 ,经过热损伤刺激的成纤维细胞 ,c junN末端激酶 (JNK)首先开始表达 ,并在 60min达到高峰 ,能持续至 180min。细胞外信号调节激酶(ERK)的表达高峰在伤后 3 0min,随后迅速消退。损伤早期 ,成纤维细胞内caspase3阳性标记的细胞数较少 ,至伤后 60min ,caspase3的表达明显增强。研究表明 。To explore the activities of signal transduction pathway involved in heat-stressed fibroblast in vitro. Human dermal fibroblasts were cultured in Dulbecco′s modified Eagle′s medium with 5% calf serum at 5%CO 2 in a water-saturated atmosphere. Cultured cells were heated to 45℃ for 10min. Western blotting was used to detect ERK1/2 and JNK expression, and the expression of caspase-3 protein was observed by immunoflurescence technique. The results showed that the MAPKs signal transduction pathway was activated in heat-stressed fibroblasts. The expression of JNK reached the peak at 60min, then maintained up to 180min. The expression of ERKs peaked at 30min, then lowered. The expression of caspase-3 was weak at 30min after heat-stress, and became evidently strong at 60min. The signal pathway of ERKs and JNK played important roles in the changes in biologic characteristics of fibroblasts after heat-stress.

关 键 词：丝裂素活化蛋白激酶 创伤与损伤 信号传递 成纤维细胞 

分 类 号：R644[医药卫生—外科学] R329.25[医药卫生—临床医学]