氨磷汀对氢醌诱导细胞凋亡的保护作用

The protective effect of amifostine on hydroquinone-induced apoptosis in bone marrow

目的 研究体外培养条件下氨磷汀对氢醌诱导的骨髓单个核细胞凋亡的影响 ,探讨氨磷汀对苯骨髓毒性的抑制作用。方法 分离人骨髓单个核细胞 ,分成空白对照组、氨磷汀组、氨磷汀+氢醌组和氢醌组。培养后 ,采用HT荧光染色观察形态学变化 ,DNA片段的琼脂糖凝胶电泳检测细胞凋亡 ,流式细胞术ANNEXINV FITC加碘化丙啶 (PI)双染定量检测不同时间细胞凋亡率和坏死率的变化。结果 培养 10h后氢醌组可见大量的凋亡细胞 ,氨磷汀加氢醌组凋亡细胞明显减少 ,差异有显著性 (P <0 .0 1)。培养 8～ 12h ,氨磷汀加氢醌组细胞凋亡率明显低于氢醌组 ,差异有显著性 (P <0 .0 1) ,培养 18～ 4 8h ,氨磷汀加氢醌组的细胞坏死率明显低于氢醌组 ,差异有显著性 (P <0 .0 5 )。结论 氨磷汀能抑制氢醌诱导的骨髓单个核细胞凋亡和减少坏死。

Objective To evaluate the protective effect of amifostine on hydroquinone-induced apoptosis of bone marrow mononuclear cells in vitro. Methods The mononuclear cells were separated and divided into four groups:blank control,amifostine group,hydroquinone group,amifostine + hydroquinone group.The cell apoptotic rate was examined in separated group at different time point,and apoptosis was detected by HT stain,then cell morphology was observed under fluorescent microscope and DNA fragments was tested by agarose gel electrophoresis.In addition,apoptotic and necrotic rate was detected by flow cytometer. Results After 10 hour culture,DNA ladder was detected in the hydroquinone group,but not in other groups.The apoptotic rate was not significantly different between amifostine group and blank control group at different culture time(P>0.05).After 8-12 hour culture,the apoptotic rate in amifostine+hydroquinone group was significantly lower than that in the group of hydroquinone alone(P<0.01).After 18-48 hour culture,the necrotic rate in amifostine+hydroquinone group was lower than that in the group of hydroquinone alone(P<0.05). Conclusion Amifostine can protect cell from hydroguinone-induced bone marrow damage through inhibition on cell apoptosis,and decrease in cell necrosis.