雷公藤单体T_(10)对Aβ1-42所致PC12细胞凋亡的抑制作用  

Effect of Chinese herb Tripterygium wilfordii Hook F monomer triptolide on apoptosis of PC12 cells induced by Aβ1-42

作　　者：顾明[1] 周慧芳[1] 薛冰[1] 牛东滨[1] 何其华[1] 王晓民[1] 

机构地区：[1]北京大学神经科学研究所,北京100083

出　　处：《生理学报》2004年第1期73-78,共6页Acta Physiologica Sinica

基　　金：This work was supported by the National Basic Research Priorities Programme of China(G1999054008);the National Natural Science Foundation of China(3027 1494).

摘　　要：阿尔茨海默病(Alzheimer’s disease,AD)是发病率最高的中枢神经系统退变性疾病。目前AD的病因不清,亦无有效的防治手段,其重要的原因是尚无适宜的AD模型。因此,本实验首先建立了PC12细胞系β淀粉样蛋白(β—amyloid,Aβ)细胞损伤模型,在此基础上,探讨了中药免疫抑制剂雷公藤单体T10对细胞的保护作用及其机制。首先用不同浓度的Aβ(5×10～4、5×10-3 、5×10-2 、5×10-1、5、50 μmol/L)与PC12细胞共孵育48 h,用MTT法检测细胞存活率。选取Aβ致使细胞存活率降低的浓度(0.5、5、50 μmol/L)与PC12细胞共孵育48 h,通过流式细胞仪检测凋亡细胞百分比。用1×10-11mol/L的T10预孵育PC12细胞48 h后,加入50μmol/L Aβ共孵育48 h,亦用流式细胞仪检测凋亡细胞百分比,激光共聚焦显微镜检测细胞内钙离子浓度变化。结果显示,Aβ的浓度在50 μmol/L时可使细胞存活率降低至55.1％,凋亡细胞比例显著增加,而1×10-11mol/L的T10可明显降低50μmol/L,Aβ诱导的PC12细胞死亡。50μmol/L。Aβ可促进PC12细胞胞外钙离子内流,1×10-11mol/L的T10对Aβ诱导的胞外钙离子内流有抑制作用。这些观察结果表明T10对Aβ导致的PC12细胞损伤具有明显的保护作用,其机制可能与抑制Aβ诱导的胞内钙离子浓度升高和细胞凋亡有关。Recent studies indicate that β-amyloid (Aβ) is the key factor to cause neuronal degeneration in Alzheimer's disease (AD). In the present study, we set up an Aβ induced PC 12 cell damage modle and studied the protective effect and related mechanisms of T10, monomer extracted from Chinese herb Tripterygium wilfordii Hook F. PC12 cells were treated with different concentrations of Aβ (5× 10 -4, 5× 10 -3 5×10 -2, 5× 10 -1, 5, 50 μmol/L) for 48 h, cell viability was detected by MTT conversion. The apoptotic rate of PC12 cells was quantitatively determined using FACS assay. After PC12 cells were treated with 1×10-11 mol/L T10 for 48 h and then co-treated with 50 μmol/L Aβ for 48 h, the apoptotic rate and the change in intracellular Ca2+ concentration of PC12 cells were analyzed by FACS assay and confocal, respectively. It was found that 5 μmol/L Aβ decreased the cell viability to 66.3% and 50 μmol/L Aβ decreased it to 55.1%, significantly different from that of the control group. After treatment with 50 μmol/L Aβ for 48 h, the apoptotic rate of PC 12 cells increased obviously. The apoptotic rate was 5.37% in the control group, while after treatment with 0.5, 5 and 50 μmol/L Aβ for 48 h, the apoptotic rate of PC 12 cells went up to 10.19%, 8.02% and 16.63%, respectively. At the same time, the concentration of intracellular Ca2+ increased greatly after treatment with 50 μmol/L Aβ for 48 h. At the concentration of 1×10-11 mol/L T10 remarkably inhibited the apoptosis induced by 50 μmol/L Aβ. In the naive group, the apoptotic rate was 4.83%. The apoptotic rate went up to 17.24% after treatment with 50 μmol/L Ap for 48 h. After co-treatment with 1×10-11 mol/L T10 and 50 μmol/ L Aβ, the apoptotic rate decreased to 8.91%, significantly different from that of the control group. At the same time, at the concentration of 1× 10 -11 mol/L T10 remarkably inhibited the increase of intracellular Ca2+ concentration induced by Aβ. The results indicate that T10 has obvious protective effect on PC12 cells, w

关 键 词：阿尔茨海默病 Β-淀粉样蛋白 雷公藤单体T10 神经保护 

分 类 号：R285[医药卫生—中药学]