丁基苯酞对帕金森病细胞模型保护作用的初步研究

The preliminary study on the protective effects of dl-3-n-butylphthalide on a parkinsonian cell model

目的探讨丁基苯酞(dl-3-n-Butylphthalide,NBP)对鱼藤酮诱导的帕金森病细胞模型的保护作用及其机制。方法分别使用终浓度为0.1、1、10、100μM NBP和溶剂二甲基亚砜(DMSO)预处理SH-SY5Y细胞24h后,加入终浓度为200nM的鱼藤酮处理24h建立多巴胺能细胞损伤模型,观察各组细胞形态,采用四甲基偶氮唑盐(MTT)比色法检测细胞活性,流式细胞术检测细胞凋亡率(Annexin V-FITC/PI)、线粒体膜电位(JC-1)、细胞内活性氧水平(DCFH-DA)。结果200nmol/L鱼藤酮处理SH-SY5Y细胞24h能够诱导细胞活性下降和细胞凋亡,NBP预处理后SH-SY5Y细胞存活率明显升高,细胞凋亡率降低,线粒体膜电位显著升高(P<0.05),细胞内活性氧水平显著降低(P<0.05),且随NBP浓度的增加对SH-SY5Y细胞的保护作用增强。结论NBP对鱼藤酮诱导的SH-SY5Y细胞损伤具有良好的保护作用,线粒体保护可能是其作用机制之一。

Objective To explore whether dl-3-n-Butylphthalide（NBP） has protective effects on a parkinsonian cell model induced by rotenone and the probable underlying mechanisms.Methods Rotenone was administrated into the cultured human neuroblastoma SH-SY5Y cells to establish a cell model of Parkinson＇s disease.The cells were pretreated with 0.1 μM,1 μM,10 μM and 100 μM NBP for 24 hours prior to rotenone administration.The cell morphology was observed,cell viability was detected by MTT,Annexin V-FITC/ propidium iodide staining was employed to analyze the apoptosis,mitochondrial membrane potential was detected by JC-1 staining;reactive oxygen species contents was tested by DCFH-DA staining.Results Rotenone induced cell viability loss and apoptosis in SH-SY5Y cells.NBP pretreatment attenuated the cell viability loss and apoptosis induced by rotenone.As compared with rotenone group,the mitochondrial membrane potential was markedly elevated（P〈0.05）,and the levels of reactive oxygen species were significantly declined（P0.05）,the protective effect of NBP was more significant at higher concentration.Conclusions NBP could ameliorate the injury induced by rotenone in SH-SY5Y cells,mitochondrial function reservation and the subsequent anti-apoptotic effect may be the underlying mechanisms in the neuroprotective effects of NBP.