鱼藤素对SH-SY5Y神经母细胞瘤细胞凋亡的诱导作用  

The apoptotic inducing effect of deguelin on SH-SY5Y cells

作　　者：吴碧娟[1,2] 姜志辉[2] 孙婧雯[1] 谭翠雯 范俣琳 丁晓艳[1] 上官信一 吴新荣[1] 

机构地区：[1]广州军区广州总医院药剂科-广州市老年慢病患者合理用药重点实验室,广东广州510010 [2]南方医科大学研究生院,广东广州510515

出　　处：《中国药理学通报》2017年第8期1136-1140,共5页Chinese Pharmacological Bulletin

基　　金：广东省科技计划项目(No 2011A080300004);广州市科技计划项目(No 201509010012)

摘　　要：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。Aim To study the apoptotic inducing effects of deguelin on SH-SY5 Y cells. Methods SHSY5 Y cells were treated with 0,0. 625,1. 25,2. 5,5,10 and 20 μmol·L-1deguelin for different time（ 24,48,72 h）; cell viability was detected by CCK-8 assay.SH-SY5 Y cells were treated with 0,8,20,50 μmol ·L-1deguelin for 24 h; light microscope and AO/EB double stained method were employed for observing the morphology and apoptotic morphology of treated cells.Apoptotic rate of treated cells was determined by flow cytometry. Cells were stained by DCFH-DA,and the whole reactive oxygen species（ ROS） was determined by flow cytometry. Spectrophotometry was employed to determine the activation degree of caspase-3. Results Deguelin inhibited cell growth in a time-and dose-dependent manner,and the IC50 value of deguelin was（ 26. 07 ± 2. 18）,（ 18. 33 ± 0. 94）,（ 12. 5 ± 1. 49）μmol·L-1when treated with 24,48,72 h respectively. After treated with 8,20,50 μmol·L-1deguelin for24 h,cell apoptotic rate,ROS and activation rate of caspase-3 increased markedly（ P〈0. 05）,all of which performed a dose related effect. Conclusion Deguelin can inhibit SH-SY5 Y cell proliferation and induce cell apoptosis,and the mechanism may be concerned with the elevated ROS and activated caspase-3.

关 键 词：鱼藤素 SH-SY5Y细胞 凋亡 活性氧 CASPASE-3 神经毒性 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R730.264[医药卫生—基础医学]