目的:在MPP^+诱导的SH—SYSY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MTF法检测Esculin对细胞存活率的影响。应用AnnexinV—FITC与H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Rhodamine123对细胞内活性氧簇(ROS)及线粒体膜电位(AWm)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AWm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).