小蜡树皮单体化合物—Esculin对MPP^+所致SH-SY5Y细胞损伤的保护作用

Esculin,an extract from Fraxinus sielboldiana blume,protects SH-SY5Y cells from MPP^+-induced cytotoxicity

目的:在MPP+诱导的SH-SY5Y细胞凋亡模型中,研究从小蜡树皮提取的单体化合物Esculin的神经保护作用。方法:应用MTT法检测Esculin对细胞存活率的影响。应用Annexin V-FITC与PI双染流式细胞仪检测Esculin对MPP+诱导的细胞凋亡的影响。此外,应用荧光染料DCFH-DA及Rhodamine 123对细胞内活性氧簇(ROS)及线粒体膜电位(ΔΨm)进行了检测。结果:在经过10-7mol/L,10-6mol/L,10-5及10-4mol/L浓度Esculin处理后,细胞存活率与MPP+处理组相比,有显著性提高。结果显示,Esculin具有显著的抗细胞凋亡作用。同时Esculin可明显改善MPP+引起的ΔΨm下降,并可以逆转MPP+诱导的ROS水平升高。结论:Esculin对MPP+诱导的细胞损伤有明显保护作用,该化合物可以作为治疗退行性神经疾病如帕金森病等的候选化合物。

Objective：To investigate the effect of Esculin extracted from Fraxinus sielboldiana blume belonging to the Oleaceae family on MPP^＋ -induced cytotoxicity in human neumblastoma SH-SYSY cells. Methods： Cell viability was tested when treated with 200 μmol/L concentrations of MPP^＋ for 48h by MTT assay. Early apoptosis, late apoptosis/necrosis were analyzed by flow cytometry using Annexin V-FITC and propidium iodide （PI） staining, respectively. Generation of reactive oxygen species （ROS） was assessed by DCFH-DA, an oxidation-sensitive fluorescent probe. To evaluate mitochondrial membrane potential （△ψm） by using flow cytometry with the fluorescent dye Rhodamine 123. Results：The MPP^＋ -induced loss of cell viability was significantly increased by Esculin treatment at the concentration of 10.7 mol/L, 10^-6 mol/L, 10^-5 and 10^-4mol/L. The protective effects of Esculin on MPP^＋ -induced cytotoxicity might be ascribed to its anti-oxidative properties by reducing ROS level and antiapoptotic effect on protection of △ψm. Conclusions：Esculin may provide a useful therapeutic strategy for the treatment of progressive neurodegenerative diseases such as Parkinson＇s disease （PD）.