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目的研究鱼藤素对SH-SY5Y细胞凋亡的诱导作用。方法鱼藤素(0、0.625、1.25、2.5、5、10、20μmol·L~(-1))处理SH-SY5Y细胞24、48、72 h后,CCK-8法测定细胞存活率。鱼藤素(0、8、20、50μmol·L~(-1))处理SH-SY5Y细胞24 h,光镜下及AO/EB双染分别观察细胞形态和凋亡形态,流式细胞术检测细胞凋亡率,DCFH-DA荧光探针法检测细胞活性氧水平,分光光度法检测caspase-3活化程度。结果鱼藤素对SH-SY5Y细胞存活率呈时间和浓度依赖性抑制作用,作用24、48、72 h的IC50值分别为(26.07±2.18)、(18.33±0.94)、(12.5±1.49)μmol·L~(-1)。鱼藤素处理24 h后,细胞凋亡率、细胞活性氧水平明显上升(P<0.05),caspase-3活化程度升高,且三者均有浓度-效应特征。结论鱼藤素可抑制SH-SY5Y细胞存活,诱导细胞凋亡,其机制可能与升高活性氧水平和活化caspase-3相关。
Objective To study the induction of deguelin on apoptosis in SH-SY5Y cells. Methods After 24h, 48h and 72h treatment of SH-SY5Y cells with deguelin (0,0.625,1.25,2.5,5,10,20 μmol·L -1), cell viability was determined by CCK-8 assay. The morphology and morphology of SH-SY5Y cells were treated with deguelin (0, 8, 20, 50μmol·L -1) for 24 h under light microscope and AO / EB double staining. Flow cytometry The apoptotic rate was detected by flow cytometry. The level of reactive oxygen species (ROS) was measured by DCFH-DA fluorescence probe and the activation of caspase-3 was detected by spectrophotometry. The results showed that deguelin inhibited the survival rate of SH-SY5Y cells in a time and concentration-dependent manner. The IC50 values of 24, 48 and 72 h were (26.07 ± 2.18), (18.33 ± 0.94) and (12.5 ± 1.49) μmol · L ~ (-1). After deguelin treatment for 24 h, the apoptotic rate and ROS level were significantly increased (P <0.05), and the activation of caspase-3 increased. All of them had concentration-response characteristics. Conclusions deguelin can inhibit the survival of SH-SY5Y cells and induce apoptosis, which may be related to the increase of reactive oxygen species level and activation of caspase-3.