紫外线照射后生成的微囊泡对成纤维细胞氧化损伤及凋亡的作用

摘要

Objective:To determine the effects of microvesicles induced by UVA and UVB irradiation on oxidative damage and apoptosis skin fibroblasts. Methods:Cultured skin fibroblasts were irradiated with UVA and UVB. Microvesicles were extracted from supernatant of cultured fibroblasts. The size and the number of microvesicles were detected by light scattering analysis technology. The microvesicles were co-incubated with normal fibroblasts. The active oxygen content was detected by reactive oxygen species ( ROS) kit and apoptosis rate was detected by flow cytometry. Results:The size and the mumber of microvesicles abstracted from fibro-blasts after UVA and UVB radiation was significantly more than those in normal fibroblasts. The active oxygen content in fibroblasts irradiated by UVA and UVB and normal fibroblasts after co-incubated with microvesicles was (52.76±1.4347), (82.60±4.082) and (85.94±6.264), and the apoptosis rates were (3.260±1.732)%, (28.94±2.430)% and (34.48±2.718)%. The cell oxidative damage and apoptosis rate can be reversed by the antioxidant. Conclusion:UVA/UVB radiation can induce skin fibroblasts to release microvesicles, which can stimulate oxidative damage and apoptosis of the cells.%目的：明确对UVA及UVB照射后皮肤成纤维细胞生成的微囊泡对成纤维细胞氧化损伤及凋亡的作用。方法：紫外线照射人皮肤成纤维细胞，提取细胞上清液中的微囊泡，利用光散射分析技术鉴定分析微囊泡的大小及数量。将紫外线照射后生成的微囊泡与正常成纤维细胞共孵育，荧光酶标仪定量检测活性氧含量，流式细胞仪检测细胞凋亡率。结果： UVA及UVB照射后皮肤成纤维细胞释放的微囊泡数量及大小明显高于正常成纤维细胞释放的微囊泡。正常纤维细胞、UVA和UVB照射后的成纤维细胞与微囊泡共孵育后活性氧荧光值分别为(52.76±1.4347)、(82.60±4.082)和(85.94±6.264)，凋亡率分别为(3.260±1.732)％，(28.94±2.430)％和(34.48±2.718)％，细胞的氧化损伤和凋亡可被抗氧化剂逆转。结论：急性中长波紫外线照射可诱导皮肤成纤维细胞释放微囊泡进一步介导细胞的氧化损伤和凋亡。