类叶升麻苷对鱼藤酮致SH-SY5Y细胞凋亡的保护作用

Protective effect of acteoside on rotenone-induced apoptosis in SH-SY5Y cells

目的探讨类叶升麻苷对鱼藤酮致多巴胺能神经元SH-SY5Y细胞凋亡的保护作用及其机制。方法采用MTT法检测细胞存活率,以荧光染料Hoechst33342染色分析细胞核的形态学变化,用流式细胞仪定量分析细胞凋亡峰,以2,′7′-二氢二氯荧光黄双乙酸钠(DCFH-DA)为标记探针检测细胞内活性氧的产生。结果①0.5μmol.L-1的鱼藤酮处理SH-SY5Y细胞48 h能引起细胞存活率的显著下降;诱导细胞发生凋亡,凋亡率达47.39%;大部分细胞胞体皱缩,突起缩短消失或断裂;染色质皱缩、浓缩、断裂及形成凋亡小体;细胞内活性氧水平上升。②预先用盐生肉苁蓉提取物类叶升麻苷(10,20或40 mg.L-1)处理细胞6 h,可提高细胞存活率;明显改善鱼藤酮引起的细胞形态学变化;流式细胞仪检测凋亡率分别降低到25.87%,23.97%,10.45%;以DCFH-DA为标记探针检测到20 mg.L-1类叶升麻苷可明显抑制鱼藤酮引起的细胞内活性氧产生。结论类叶升麻苷能抑制鱼藤酮诱导的多巴胺能神经元SH-SY5Y细胞凋亡,其神经细胞保护作用可能与降低细胞内活性氧水平有关。

Aim To investigate whether acteoside can protect the dopaminergic neurons, SH-SYSY cells,from rotenone-induced apoptosis. Methods Cell viability was analyzed by MTF assay. The neurons were stained with the fluorescent dye, Hoechst 33342, to analyze the nuclear change. Flow cytometry was used to determine neuronal apoptotic peak quantitatively. The level of intracellular reactive oxygen species （ROS） was monitored using the fluorescent probe 2＇, 7＇-dichlorofluorescin- diacetate （DCFH-DA）. Results ①SH-SY5Y cells treated with 0. 5 μmol·L^-1 rotenone for 48 h had a significantly decrease on cell viability compared with control, and the percentage of apoptosis was increased to 47.39%. The cell bodies of most cells shrinked and the dendrites of most cells became shorten or broken. condensation and cleavage be observed. The level of Meanwhile, the shrinkage, of nuclei in most cells could intracellular oxygen species was enhanced. ② The preincubation with acteoside （10,20 or 40 mg · L^-1）, one of the phenylethanoids isolated from Cistanche salsa, for 6 h enhanced the cell viability, and improved the rotenone-induced cell mor- phological change. Moreover,the percentage of apoptosis was significantly decreased to 25.87%, 23. 97% and 10. 45% in a dose-dependent manner. The enhancement of ROS induced by rotenone was inhibited by 20 mg · L^-1 acteoside. Conclusion These results demonstrate that acteoside can protect SH-SYSY cells against rotenone-induced apoptosis. The neuroprotective effect might be related with the function that acteoside can reduce the level of ROS.