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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。该方法有助于在离体情况下探讨创伤修复过程中成纤维细胞生物学特性
OBJECTIVE: To establish a model of apoptosis induced by cell scald in vitro and to study the changes of the main repair cells fibroblasts after scald. Methods Fibroblasts in DMEM medium containing 5% and 10% FCS were cultured in vitro at different temperatures (43, 45, 48 ℃) and different time (10, 30 , 40min), the control group cells were placed in 37 ℃ water bath for 30min. The scalded cells were detected by DNA gel electrophoresis, Hoeschst332 58 fluorescence staining and transmission electron microscopy, and the results were analyzed. Results In 43 ℃ and 45 ℃ water bath, the apoptosis of fibroblasts could be observed at 10, 30 and 40 min, and the best one was DMEM containing 5% fetal bovine serum at 10 min in 45 ℃ water bath obvious. 48 ℃ water bath for more than 30min, then a large number of cell necrosis. No significant apoptosis occurred in the control group. Conclusion Thermal injury can cause apoptosis of isolated fibroblasts. This method is useful for exploring the biological characteristics of fibroblasts during wound healing in vitro