三氧化二砷对兔晶体上皮细胞增殖的影响及其机制的实验研究

In Vitro Study on Inhibition of Rabbit Crystal Epithelial Cell by Arsenic Trioxide

目的:观察不同浓度的三氧化二砷作用不同时间对兔晶体上皮细胞影响及其作用机理。方法:应用不同浓度的三氧化二砷作用于兔晶体上皮细胞后,观察三氧化二砷对兔晶体上皮细胞生长状态的影响;用噻唑蓝(MTT)比色法、透射电镜技术观察其对兔晶体上皮细胞增殖的影响;应用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)及流式细胞技术检测三氧化二砷对兔晶体上皮细胞凋亡的诱导作用。结果:不同浓度的三氧化二砷作用于兔晶体上皮细胞有明显的时间和剂量依赖性。三氧化二砷作用后细胞生长明显受抑制,并出现明显的凋亡特征性改变;8μmol/L三氧化二砷作用24小时、48小时及72小时后TUNEL法可检测到细胞凋亡水平显著性升高,流式细胞仪分析显示,兔晶体上皮细胞在G1峰前出现明显的凋亡峰。结论:三氧化二砷可明显抑制兔晶体上皮细胞的生长,其机理主要是诱导兔晶体上皮细胞凋亡。

Objective：To investigate the effects and mechanisms of arsenic trioxide（As2O3）on rabbit crystal epithelial cell.Methods：Rabbit crystal epithelial cells were cultured with 0.5 to 16 μmol/L arsenic trioxide for various durations,then cell viability was measured by MTT assay.The morphological changes of the cells were examined by electron microscopy.The cell necrosis and apoptosis rates were observed by terminal deoxynucleotidyl transferase mediated nick end labeling（TUNEL）and flow cytometry.Results：The effect of arsenic trioxide on Rabbit crystal epithelial cells was dependent on the time and concentration obviously.The decrease in cell number was preceded by electron morphological changes in the treated Rabbit crystal epithelial cells that were characteristic of apoptosis,including membrane blebbing,shrunken cytoplasm,nuclear condensation and loss of adhesion.Apoptosis index was elevated by TUNEL after incubating with 8μmol/L arsenic trioxide for 1～3 days.Flow cytometry study indicated that ratio of G0/G1 phase cell was decreased and the portion of S phase cell increased following treated with As2O3.Conclusion：As2O3 could obviously inhibit the growth of Rabbit crystal epithelial cells through inducing Rabbit crystal epithelial cells apoptosis.