离体成纤维细胞热损伤模型制作的研究  

Heat injury fibroblasts: a useful model to study wound healing in vitro

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]解放军第三○四医院创伤外科研究室,北京100037 [2]第四军医大学基础部全军神经科学研究所

出　　处：《中华创伤杂志》2002年第7期391-393,共3页Chinese Journal of Trauma

基　　金：国家重点基础研究发展规划资助项目(G19990 54 2 0 4 );国家自然科学基金资助项目 ( 30 170 96 6 );国家杰出青年科学基金资助项目 ( 3952 50 2 4 );国家博士后基金资助项目 (中博基[2 0 0 1]No .14 )

摘　　要：目的　建立离体细胞烫伤诱导凋亡的模型 ,深入研究组织烫伤后主要修复细胞成纤维细胞的变化。　方法　利用体外细胞培养技术对含小牛血清 (体积分数为 5 %和 1 0 % )的DMEM培养液中的成纤维细胞进行不同温度 (43 ,45 ,48℃ )和不同时间 (1 0 ,30 ,40min)的处理 ,对照组细胞置于 37℃水浴 30min。采用DNA凝胶电泳、Hoeschst332 58荧光染色、透射电镜等技术对烫伤细胞进行检测并分析结果。　结果　在 43℃和 45℃水浴中 ,成纤维细胞孵育 1 0 ,30 ,40min均可出现细胞凋亡 ,以含体积分数为 5 %小牛血清的DMEM培养液、45℃水浴 1 0min一组最为明显。 48℃中水浴 30min以上 ,则出现大量的细胞坏死。对照组中无明显的细胞凋亡发生。　结论　热损伤能造成离体成纤维细胞发生凋亡。Objective To explore the change of reparable cells in scalded tissue and to establish a model of apoptosis due to heat injury in vitro. Methods Human dermal fibroblasts were cultured in vitro, cells in Dulbecco's modified Eagle's medium cultured in a water saturated atmosphere. Dulbecco's modified Eagle's medium was supplied with 10% or 5% calf serum. Cells were stimulated at 43℃,45℃ and 48℃ for 10 minutes, 30 minutes and 40 minutes, respectively. DNA gel electrophoresis, Hoeschst 33258 fluorescence staining and transmission electron microscope were used to determine the condition of apoptosis. In the control group, fibroblasts were incubated at 37℃ for 30 minutes. Results Fibroblast apoptosis occurred at 10, 30 and 40 minutes under water bath at 43℃ and 45℃. Fibroblasts in Dulbecco's modified Eagle's medium was supplied with 5% calf serum and stimulated at 45℃ for 10 minutes. There were a lot of apoptotic cells. However, in the cultured fibroblasts stimulated at 48℃ for more than 30 minutes, there was mass cellular necrosis, on the contrary, there were no significant apoptotic cells in the control group. Conclusions Apoptosis in fibroblasts can be successfully induced by heat injury in vitro. This model can be used to study the biological character of fibroblasts under wound healing in vitro.

关 键 词：成纤维细胞 创伤 损伤 凋亡 生物学模型 

分 类 号：R-331[医药卫生]