正常人成纤维细胞老化过程中活性氧水平的变化

The Alteration of the Level of Reactive Oxygen Species in the Aging of Normal Human Fibroblasts

目的研究正常人成纤维细胞在老化过程中细胞活性氧水平的变化。 方法取常规体外培养的原代人成纤维细胞 ,按不同传代数 (PD2 2 - 2 4和PD5 0 - 5 2 )分为两组 ,代表老化过程中两个阶段 (早期和中—晚期 ) ,用流式细胞仪检测罗丹明 12 3显示细胞活性氧水平 ,同时用β -半乳糖苷酶组化染色 ,形态观察衰老细胞 ,用群体倍增时间和流式细胞周期分析显示细胞增殖情况。 结果中—晚期细胞中衰老细胞增多 ,活性氧水平增高 ,未出现增殖阻滞。 结论正常人成纤维细胞在老化过程中细胞活性氧水平呈升高趋势。

Objective To study the alteration of the level of reactive oxygen species (ROS) in the aging of the normal human fibroblasts. Methods The primary human fibroblasts cultured under routine condition at PD 22-24 and 50-52 were taken to represent respectively two stages during aging (early phase and middle-late phase). Flow cytometry was used to detect rodamine 123 and thus to show the ROS level in the cells. Aging morphology was displayed by microscopy and β-galactosidase histochemical staining. Cell replication was assessed by population doubling time and cell cycle analysis. Results Cells with senescence morphology were increased in middle-late phase fibroblasts. The ROS level was remarkably higher in middle-late phase cells than in early phase ones, though replication arrest didnt occur at this stage yet. Conclusion The level of ROS in the normal human fibro blasts is increased in the aging progress.