目的 探讨细胞外三磷腺苷(ATP)对PC12细胞损伤作用的剂量和时间依赖性特点.方法 将对数生长期的PC12细胞分别用0.00、0.03、0.10、0.30、0.50、1.00、2.00、3.00、5.00、7.00和9.00 mmol·L-1ATP处理3h后,光学显微镜下观察细胞形态的变化,甲氮甲唑蓝(XTT)比色法检测细胞死亡率,锥虫蓝染色检测细胞存活率.将对数生长期的PC12细胞分别在7.00 mmol·L-1ATP作用0、1、2、3、6、12、24 h时,用XTT比色法检测细胞死亡率.结果 (1)XTT比色法检测结果显示,0.03、0.10、0.30、0.50、1.00和2.00 mmol·L-1 ATP组PC12细胞死亡率与0.00 mmol · L-1 ATP组比较差异无统计学意义(P＞0.05).随着浓度升高,ATP诱导PC12细胞死亡作用逐渐增强,3.00、5.00、7.00和9.00 mmol·L-1 ATP明显增加细胞的死亡率,与0.00 mmol·L-1 ATP组比较差异有统计学意义(P＜0.05).(2)锥虫蓝染色检测结果显示,0.03、0.10、0.30、0.50和1.00 mmol·L-1ATP组细胞存活率与0.00 mmol·L-1 ATP组比较差异无统计学意义(P ＞0.05)；2.00、3.00、5.00、7.00和9.00 mmol· L-1ATP组细胞存活率与0.00 mmol · L-1 ATP组比较差异有统计学意义(P＜0.05,P＜0.01).(3)XTT法检查7.00 mmol·L-1ATP处理细胞死亡率显示,1、2h组与0h组比较差异有统计学意义(P＜0.05),3、6、12、24h组与0h组比较差异更显著(P＜0.01),3、6、12、24h组之间比较差异无统计学意义(P＞0.05).结论 ATP对PC12细胞毒性作用具有剂量和时间依赖性特点.%Objective To explore the dose and time dependence feature of extracellular adenosine triphosphate (ATP) induced damage in PC12 cells. Methods The exponential growth phase cells were induced by 0.00,0.03,0. 10,0.30,0. 50, 1.00,2.00,3.00,5.00,7.00 and 9.00 mmol·L-1 ATP. The following tests were done when PC12 cells exposed to ATP for 3 hours. Cells morphology were observed by microscopic inspection. Cell death rate was detected by 2,3-bis(2-methoxy-4-ni-tro-5-sulfophenyl)-5-[ ( phenylamino) carbonyl ]-2H-tetrazolium hydroxide ( XTT). The cell survival rale was detected by trypan blue staining. The exponential growth phase cells were exposed to 7.00 mmol·L-1 ATP for 0,1,2,3,6,12,24 hours respectively, and the cell death rate of the corresponding time point was detected by XXT. Results (1) Compared with 0.00 mmol·L-1 ATP group, there was no significant difference in mortality rate in 0. 03,0. 10, 0. 30,0. 50,1. 00 and 2.00 mmol·L-1 ATP groups( P >0. 05). But with increasing of ATP concentration,the mortality rate increased gradually; there was significantly difference in cells mortality rate in 3.00,5.00,7.00 and 9.00 mmol·L-1 ATP groups compared with 0.00 mmol·L-1 group(P<0. 05). (2) Compared with 0.00 mmol·L-1 ATP group,there was no significant difference in cells survival rate in 0. 03,0.10,0. 30,0.50 and 1.00 mmol· L-1 ATP groups(P>0.05) ;there was significant difference in cells survival rate in 2.00,3.00,5.00,7.00 and 9.00 mmol·L-1 ATP groups( P <0. 05,P <0. 01). (3 ) The result of the cells which were exposed to 7.00 mmol·L-1 ATP for 0,1,2,3,6,12,24 hours showed that, there was significant difference in cell death rate in 1 hour group and 2 hours groups compared with 0 hour group(P<0. 05) ;compared with 0 hour group,and the differences in 3,6,12,24 hours group were more significantly(P <0. 01) ,and the cell death rate in 3,6,12,24 hours groups had no obviously difference each other( P >0.05). Conclusion The toxicity of ATP to PC12 cells has the dose and time dependence feature.
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