CAJ | 학술논문

目的用多巴胺诱导的SH-SY5Y细胞凋亡模型,研究从小蜡树皮提取的单体化合物liriodendrin的神经保护作用.方法应用MTT法检测liriodendrin对细胞存活率的影响.应用Annexin V-FITC与PI双染流式法检测liriodendrin对多巴胺诱导的细胞凋亡的影响.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(⊿ψm)进行了检测.此外,应用RT-PCR方法,对细胞内P53基因的转录水平进行了检测.结果在经过10-8,10-7,10-6,10-5及10-4 mol·L-1 浓度liriodendrin处理后,细胞存活率与多巴胺处理组相比有显著性提高.流式细胞术结果显示liriodendrin具有显著的抗细胞凋亡作用.同时liriodendrin可明显改善多巴胺引起的⊿ψm下降,并可以逆转多巴胺诱导的ROS水平升高.此外,liriodendrin还可以降低多巴胺引起的P53基因转录水平的升高.结论 Liriodendrin对多巴胺诱导的细胞损伤有明显保护作用.推测主要机制可能与其调节细胞氧化应激反应及细胞凋亡的信号转导通路有关,提示该化合物可以作为治疗退行性神经疾病如帕金森氏病等的候选化合物.
목적 용다파알유도적SH-SY5Y세포조망모형,연구종소사수피제취적단체화합물liriodendrin적신경보호작용.방법 응용MTT법검측liriodendrin대세포존활솔적영향.응용Annexin V-FITC여PI쌍염류식법검측liriodendrin대다파알유도적세포조망적영향.응용형광염료DCFH-DA급Rhodamine 123대세포내활성양족(ROS)급선립체막전위(⊿ψm)진행료검측.차외,응용RT-PCR방법,대세포내P53기인적전록수평진행료검측.결과 재경과10-8,10-7,10-6,10-5급10-4 mol·L-1 농도liriodendrin처리후,세포존활솔여다파알처리조상비유현저성제고.류식세포술결과현시liriodendrin구유현저적항세포조망작용.동시liriodendrin가명현개선다파알인기적⊿ψm하강,병가이역전다파알유도적ROS수평승고.차외,liriodendrin환가이강저다파알인기적P53기인전록수평적승고.결론 Liriodendrin대다파알유도적세포손상유명현보호작용.추측주요궤제가능여기조절세포양화응격반응급세포조망적신호전도통로유관,제시해화합물가이작위치료퇴행성신경질병여파금삼씨병등적후선화합물.
Aim To investigate the effect of liriodendrin,an extract from Fraxinus sielboldiana blume belonging to the Oleaceae familv,on dopamine-induced cytotoxicity in human neuroblastoma SH-SY5Y cells.Methods Cell viability was processed when treated with 50 μmol·L-1 of dopamine for 24 h by MTT assay.Early apoptosis,late apoptosis/necrosis were analyzed by flow cytometry using Annexin V-FITC and propidium iodide(PI)double-staining,respectively. Generation of reactive oxygen species (ROS) was assessed by DCFH-DA,an oxidation-sensitive fluorescent probe.To evaluate mitochondrion membrane potential (⊿ψm) using flow cytometry with the fluorescent dye Rhodamine 123.The transcriptional level of P53 Was studied using RTPCR.Results The dopamine-induced loss of cell viability was significantly attenuated by liriodendrin treatment at the concentration of 10-8,10-7,10-6,10-5 and 10-4 mol·L-1.The protective effects of liriodendrin(10-7,10-6 and 10-5 mol·L-1)on dopamine-induced cytotoxicity mav be ascribed to its anti-oxidative properties by reducing ROS level and anti-apoptotic effect via protection of ⊿ψm.In addition.the effect of liriodendrin may involve the P53 pathway in apoptosis.Conclusion Liriodendrin may provide a useful therapeutic strategy for the trealment of neurodegenerative diseases such as Parkinson's disease(PD)