白果内酯在鱼藤酮处理的PC12细胞中抑制α-共核蛋白蛋白寡聚体形成的实验研究  

Bilobalide restrains the oligomer of α-synuclein in PC12 cells treated with rotenone

作　　者：曾志芬[1,2] 井秀娜[1] 梁嫣然[1] 杨炼红[1] 刘军[1] 肖颂华[1] 陶恩祥[1] 

机构地区：[1]中山大学孙逸仙纪念医院神经科老年神经专科,广州510120 [2]佛山市南海区第二人民医院神经内科

出　　处：《中华脑科疾病与康复杂志（电子版）》2013年第1期25-29,共5页Chinese Journal of Brain Diseases and Rehabilitation（Electronic Edition）

摘　　要：目的研究白果内酯对鱼藤酮诱导的帕金森病细胞模型是否具有保护作用,对α-共核蛋白(α-synuclein)的表达及聚集是否有影响。方法使用鱼藤酮对嗜铬细胞瘤株PC12细胞进行处理,建立α-synuclein蛋白高表达的帕金森病细胞模型,白果内酯进行干预,采用四甲基偶氮唑盐法(MTT法)检测细胞活性,流式细胞术检测细胞凋亡,Western blotting法检测α-synuclein蛋白的表达。数据以均数±标准差(x珋±s)表示,应用SPSS 16.0统计软件,多组间比较采用单因素方差分析,两组间独立样本的比较采用LSD-t检验,P<0.05认为差异有统计学意义。结果 1.6μmol/L鱼藤酮组细胞活性显著低于对照组(t=17.422,P<0.01),细胞凋亡率和α-synuclein蛋白量高于对照组(t=9.141,t=8.392;P均<0.01);10μmol/L及50μmol/L白果内酯组细胞活性均高于鱼藤酮组(t=4.257,t=6.501;P均<0.01),且高剂量者细胞活性更优(t=2.933,P=0.043);10μmol/L及50μmol/L白果内酯组细胞凋亡率较鱼藤酮组低(t=4.482,t=4.488,P均<0.01),α-synuclein蛋白的表达量低于鱼藤酮组(t=8.349,t=9.028,P均<0.01),但两剂量间二者均无统计学差异(t=0.831,P=0.45;t=2.178,P=0.095);实验中所显影的α-synuclein蛋白分子量在57 kD左右。结论白果内酯对鱼藤酮诱导的PC12细胞损伤有保护作用,该保护作用可能通过抑制α-synuclein蛋白寡聚物的形成来实现。Objective To investigate the effects of bilobalide on rotenone-mediated cell damage and expression of α-synuclein. Methods PC12 cells were treated with rotenone to create a Parkinson＇s disease cell model with over expression of α-synuelein. After co-incubation with balobalide, cell viability was determined by MTF assay, the expression of α-synuclein was measured by Western blotting, cell apoptosis was assessed by flow cytometry （ FCM ）. The results were presented as mean ＋ SD, using SPSS 16. 0. Overall difference among 4 groups were determined by analysis of variance（ANOVA）. Specific pair-wise differences were determined using LSD-t-test. Differences were considered significant when P 〈 0. 05. Results Compared with control, the cell viability was significantly decreased （ t = 17.422, P 〈 0. 01 ）, cell apoptosis rate and a-synuelein expression was significantly increased （ t = 9. 141, t = 8. 392 ; both P 〈 0. 01 ） in rotenone （1.6 p.mol/L ）group. Compared with rotenone group, after co-incubation with bilobalide of 10 μmol/L or 50 μmol/L, cell viability was significantly increased（ t = 4. 257, t = 6. 501, both P 〈 0. 01 ）, and the latter got better viability than the former（ t = 2. 933, P = 0. 043 ） ; cell apoptosis rate （ t = 4. 482, t = 4. 488, both P 〈 0. 01 ） and α-synuclein expression （ t = 8. 349, t = 9. 028, both P 〈 0. 01 ） was significantly decreased, but no difference was seen between the two groups （ t = 0. 831, P = 0. 45 ; t = 2. 178,P = 0. 095 ）.The α-synuclein protein detected had a molecular weight of 57 kD, and was therefore an oligomer or an aggregated form of the protein. Conchmion Bilobalide may restrain the expression and aggregation ofα-synuclein and have a protective effect on PC12 cells treated with rotenone.

关 键 词：突触核蛋白 鱼藤酮 帕金森病 阿尔茨海默病 白果内酯 

分 类 号：R285.5[医药卫生—中药学]