摘要
目的 用三丁基过氧化氢 (t- BHP)诱导 WI- 38细胞衰老 ,建立体外细胞衰老的模型。 方法 用t- BHP(10 0μmol/ L )作用于 WI- 38细胞 ,从 30代开始 ,隔代作用 1次 ,每次 1h,共 4次 ,诱导细胞衰老。从光镜、透射电镜观察细胞形态及细胞超微结构 ,流式细胞术分析细胞周期 ,计算 G1 期细胞比例 ,对细胞进行 SA-β-半乳糖苷酶染色 ,观察 t- BHP对上述指标的影响。 结果 t- BHP(10 0μm ol/ L )作用 4次后 ,WI- 38细胞出现衰老的特征 ,细胞停止增殖分裂 ,体积增大 ,胞体变平 ,次级溶酶体增多 ,同时 G1 期细胞比例增加 ,β-半乳糖苷酶染色阳性细胞数增加。 结论 t- BHP可诱导 WI- 38细胞衰老 。
Objective\ To induce WI\|38 cells senescence by tertbutyl\|hydroperoxide(t\|BHP) and establish a model of premature senescence in vitro.\ Methods\ From 30 to 38 population doubling(PD), WI\|38 cells were exposed to 100 μmol/L t\|BHP for 1 h at every two PD.\ Then the changes of cell shape and ultrastructure were observed by microscope and transmission electron microscopy, and the percent of cells in the G\-1 phase and SA\|β\|galactosidase activity were measured with flow cytometry and cytochemistry staining, respectively.\ Results\ After four attack with 100 μmol/L t\|BHP, the cell proliferation and division in WI\|38 cells was ceased, which showed cell became larger and flat and cell growth was arrested in G\-1 phase.\ In addition, the number of secondary lysosome and activity of β\|galactosidase were increased in WI\|38 cells, which is the characteristic of premature senescence.\ Conclusion\ t\|BHP can induce senescence in WI\|38 cells and it can be acted as a model of premature senescence in vitro.\;
出处
《福建医科大学学报》
2003年第1期19-22,T001,共5页
Journal of Fujian Medical University
基金
福建省自然科学基金资助项目 ( C0 2 10 0 17)
