苦参碱抑制神经母细胞瘤SK-N-SH细胞增殖作用研究

目的: 观察苦参碱对人神经母细胞瘤SK-N-SH细胞增殖的影响，并探讨其作用机制。 方法: 取对数生长期SK-N-SH细胞，用磺酰罗丹明B（SRB）法检测苦参碱（0.5，1.0，2.0 g·L^-1）作用24，48，72 h后 SK-N-SH细胞的增殖抑制率，流式细胞仪检测细胞凋亡率和细胞周期，Western blot 检测半胱天冬氨酸蛋白酶（Caspase）-9和Caspase-3蛋白的表达。 结果: 苦参碱对 SK-N-SH细胞增殖具有明显抑制作用，且表现出剂量和时间依赖性。0.5，1.0，2.0 g·L^-1苦参碱作用24，48，72 h后SK-N-SH细胞的抑制率分别为（19.06±3.17）%，（30.18±3.02）%，（38.55±6.12）%;（45.12±4.02）%，（60.45±5.51）%，（71.38±7.91）%;（58.91±4.36）%，（73.44±8.17）%，（88.37±4.57）%。流式细胞仪检测对照组及各浓度组凋亡率分别为（2.45±0.49）%，（12.56±2.21）%，（19.44±4.32）%，（30.12±3.35）%，与对照组比较，苦参碱组的凋亡率均显著性增高（P<0.001）;细胞周期结果显示苦参碱作用SK-N-SH细胞48 h后G₀/G₁期细胞明显增多（P<0.01），S期细胞明显减少（P<0.01）。苦参碱可显著上调Caspase-9和Caspase-3蛋白表达（P<0.01，P<0.01）。 结论: 苦参碱对神经母细胞瘤SK-N-SH细胞的增殖具有抑制作用，这种作用与其阻滞细胞周期和激活Caspase-9和Caspase-3表达进而诱导产生细胞凋亡有关。

Anti-proliferative Effects of Matrine on Human Neuroblastoma SK-N-SH Cells

Objective: To investigate anti-proliferative effects of matrine on human neuroblastoma SK-N-SH cells and its mechanism. Method: Effects of matrine(0.5,1.0,2.0 g· L^-1) on SK-N-SH cell proliferation were tested by SRB assay. Cell cycle and apoptosis of SK-N-SH were measured by flow cytometry. The levels of caspase-3 and caspase-9 within the cells were analyzed by Western blot. Result: Matrine inhibited the proliferation of SK-N-SH cells in a dose-and time-dependent manner.The inhibition rate was(19.06±3.17)%,(30.18±3.02)%,(38.55±6.12)%;(45.12±4.02)%,(60.45±5.51)%,(71.38±7.91)%;(58.91±4.36)%,(73.44±8.17)%,(88.37±4.57)% after treated with matrine by 0.5,1.0,2.0 g· L^-1 in 24 h,48 h and 72 h,respectively. Flow cytometry apoptosis in the control group and each concentration group were (2.45±0.49)%,(12.56±2.21)%,(19.44±4.32)%,(30.12±3.35)%,compared with the control group,the matrine-treated cells showed increased apoptosis rate (P<0.001),increased percentage arrested in G₀/G₁ phase (P<0.01) and decreased S phase cells (P<0.01).Matrine significantly increased the expression of caspase-3 and caspase-9 of SK-N-SH cells. Conclusion: Matrine can inhibit the proliferation of SK-N-SH cell and the mechanism may be correlated to cell cycle arrest and apoptosis induced by activation of caspase-3 and caspase-9.