苦参碱对人神经母细胞瘤SH-SY5Y细胞的作用机制

Mechanism of Matrine on Human Neuroblastoma Cell SH-SY5Y

目的探讨苦参碱对人神经母细胞瘤(NB)SH-SY5Y细胞的作用机制。方法取对数生长期的细胞,分为对照组(含细胞、无苦参碱)及药物处理组(苦参碱质量浓度为2.0 g.L-1,对细胞的作用时间分别为16 h、24 h、32 h)共4组,每组5个复孔。应用四甲基偶氮唑蓝(MTT)比色法检测NB SH-SY5Y细胞增殖抑制率;流式细胞仪(FCM)检测细胞凋亡率;比色法检测Caspase-8的相对活性。采用SPSS 16.0统计软件进行单因素方差分析。结果 MTT检测对照组及各时间点药物处理组增殖抑制率分别为(4.98±1.20)%、(11.01±0.85)%、(15.22±0.71)%和(22.31±1.45)%;FCM法检测对照组及各时间点药物处理组细胞的凋亡率分别为(5.23±1.19)%、(10.74±1.65)%、(14.00±0.98)%和(17.81±1.06)%;比色法测定各组Caspase-8的相对活性分别为(4.25±1.00)%、(10.69±1.10)%、(14.80±1.44)%和(19.80±0.97)%;以上检测各组间比较差异均有统计学意义(Pa<0.05)。结论苦参碱可抑制人NB SH-SY5Y细胞增殖,并可通过上调Caspase-8的活性诱导NB SH-SY5Y细胞凋亡,其作用随时间的延长逐渐增强。

Objective To investigate the mechanism of matrine on human neuroblastoma cell SH -SYSY（ NB SH -SYSY） through the experiments. Methods The cells at logarithm growth phase were chosen and randomly divided into control group without matrine and 3 time groups,in which the action time of matrine were 16 h ,24 h and 32 h,respectively. Five duplicating holes were set up on each group and the matrine concentrations were 2.0 g L-1. The proliferation inhibition rate was detected by methyl thiazolyl tetrazolium, the apoptosis rate by flow cytometry （FCM）, and the relative Caspase -8 activity by colorimetric assay. Statistical analysis was performed by using SPSS 16.0 soft- ware for One - Way ANOVA. Results The proliferation inhibition rates of all the groups were （ 4.98 ± 1.20 ） % , （ 11. O1 ± 0.85 ） % , （ 15.22 ＋ O. 71 ） % and （22.31 ＋ 1.45 ） %, respectively ; The apoptosis rates were （5.23 ± 1.19） %, （ 10.74 ± 1.65 ） %, （ 14. O0 ± 0.98 ） % and （ 17.81 ± 1.06） %, respectively ; The relative Caspase - 8 activity were （ 4.25 ± 1.00 ） %, （ 10.69 ± 1.10 ） %, （ 14.80 ± 1.44 ） % and （ 19.80 ± 0.97 ） % , respectively. Significant differences were found in between all the groups （ Pl 〈 O. 05 ）. Conclusions Matrine can restrain human neuroblastoma SIt - SY5Y cells proliferation and induce them apoptosis by increasing the Caspase - 8 activity, whose effects strengthened following with the extending of time.