caspase-3在多塞平诱导大鼠神经细胞凋亡中的作用

Effect of caspase-3 on doxepin-induced apoptosis in rat neurons

目的 探讨caspase-3在多塞平诱导大鼠神经细胞凋亡中的作用.方法 将PC12细胞接种于培养板中,采用随机数字表法分为4组（n=10）,正常对照组（C组）不做任何处理,继续培养24 h;多塞平组（D组）培养液中加入多塞平,终浓度为120 μmol/L,孵育24 h;caspase-3抑制剂Z-DEVD-FMK组（Z组）培养液中加入Z-DEVD-FMK,终浓度为10 μmol/L,孵育24 h;多塞平＋Z-DEVD-FMK组（DZ组）培养液中加入多塞平和Z-DEVD-FMK,终浓度分别为120和10 μmol/L,孵育24 h.各组处理结束后,采用MTT法检测细胞存活率,倒置显微镜下观察细胞形态并采用流式细胞术检测细胞凋亡率.结果 与C组比较,D组和DZ组细胞存活率降低,凋亡率升高（P＜0.01）,Z组上述指标差异无统计学意义（P＞0.05）;与D组比较,DZ组细胞存活率升高,凋亡率降低（P＜0.01）.DZ组细胞形态学改变较D组减轻.结论 caspase-3可能介导了多塞平诱发的大鼠神经细胞凋亡.

Objective To investigate the effect of caspases-3 on doxepin-induced apoptosis in rat neurons.Methods The PC12 cells seeded in culture plates were randomly divided into 4 groups （n =10 each） using a random number table：normal control group （group C）;doxepin group （group D）;caspase-3 inhibitor Z-DEVD-FMK group （group Z）;doxepin ＋ Z-DEVD-FMK group （group DZ）.In group C,the cells were continuously incubated for 24 h.In group D,doxepin was added with the final concentration of 120 μmol/L,and the cells were continuously incubated for 24 h.In group Z,Z-DEVD-FMK was added with the final concentration of 10 μmol/L,and the cells were continuously incubated for 24 h.In group DZ,doxepin and Z-DEVD-FMK with the final concentrations of 120 and 10 μmol/L,respectively,were added,and the cells were continuously incubated for 24 h.After 24 h of incubation,the cell viability was detected by methyl thiazolyl tetrazolium assay,the cell morphology was observed under inverted microscope,and the neuronal apoptosis was measured by flow cytometry.Apoptosis rate was calculated.Results Compared with group C,the cell viability was significantly decreased,and apoptosis rate was increased in D and DZ groups （P〈0.01）,and no significant change was found in the parameters mentioned above in group Z （P 〉 0.05）.Compared with group D,the cell viability was significantly increased,and apoptosis rate was decreased in group DZ （P〈 0.01）.The morphological changes were significantly mitigated in group DZ as compared with group D.Conclusion Caspases-3 may mediate doxepin-induced apoptosis in rat neurons.