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雷公藤单体T_(10)对A#beta#1-42所致PC12细胞凋亡的抑制作用

顾明, 周慧芳, 薛冰, 牛东滨, 何其华, 王晓民

北京大学神经科学研究所.北京 100083

摘要

阿尔茨海默病(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,该实验首先建立了PC12细胞系#beta#淀粉样蛋白(A#beta#)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T_(10)对细胞的保护作用及其机制。首先用不同浓度的A#beta#(5×10~(-4)、5×10~(-3)、5×10~(-1)、5、50 #mu#mol/L)与PC12细胞共孵育48h,用MTT法检测细胞存活率。选取A#beta#致使细胞存活率降低的浓度(0.5、5、50 #mu#mol/L)与PC12细胞共孵育48h,通过流式细胞仪检测凋亡细胞百分比。用1×10~(-11) mol/L的T_(10)预孵育PC12细胞48h后,加入50 #mu#mol/L A#beta#共孵育48h,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,A#beta#的浓度在50 #mu#mol/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~(-11) mol/L的T_(10)可明显降低50 #mu#mol/L A#beta#诱导的PC12细胞死亡。50 #mu#mol/L A#beta#可促进PC12细胞胞外钙离子内流,1×10~(-11) mol/L的T_(10)对A#beta#诱导的胞外钙离子内流有抑制作用。这些观察结果表明T_(10)对A#beta#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A#beta#诱导的胞内钙离子浓度升高和细胞凋亡有关。

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

Effect of Chinese herb Tripterygium wilfordii Hook F monomer triptolide on apoptosis of PC12 cells induced by A#beta#1-42

Gu Ming, Zhou Huifang, Xue Bing, Niu Dongbin, He Qihua, Wang Xiaomin

Neuroscience Research Institute,Peking University.Beijing 100083

Abstract

It was found that 5 #mu#mol/L A#beta# decreased the cell viability to 66.3% and 50 #mu#mol/L A#beta# decreased it to 55.1%, significantly different from that of the control group. After treatment with 50 #mu#mol/L A#beta# for 48h, the apoptotic rate of PC12 cells increased obviously. The apoptotic rate was 5.37% in the control group, while after treatment with 0.5, 5 and 50 #mu#mol/L A#beta# for 48h, the apoptotic rate of PC12 cells went up to 10.19%, 8.02% and 16.63%, respectively. At the same time, the concentration of intracellular Ca~(2+) increased greatly after treatment with 50 #mu#mol/L A#beta# for 48h. At the concentration of 1×10~(-11) mol/L T_(10) remarkably inhibited the apoptosis induced by 50 #mu#mol/L A#beta#. In the naive group, the apoptotic rate was 4.83%. The apoptotic rate went up to 17.24% after treatment with 50 #mu#mol/L A#beta# for 48 h. After co-treatment with 1×10~(-11) mol/L T_(10) and 50 #mu#mol/L A#beta#, 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 T_(10) remarkably inhibited the increase of intracellular Ca~(2+) concentration induced by A#beta#. The results indicate that T_(10) has obvious protective effect on PC12 cells, which may be related to the inhibition of the cell apoptosis and increment of intracellular Ca~(2+) concentration induced by A#beta#.

Key words: Alzheimer's disease;P-amyloid;T_(10);neuroprotective effect

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引用本文：

顾明, 周慧芳, 薛冰, 牛东滨, 何其华, 王晓民. 雷公藤单体T_(10)对A#beta#1-42所致PC12细胞凋亡的抑制作用[J]. 生理学报 2004; 56 (1): .

Gu Ming, Zhou Huifang, Xue Bing, Niu Dongbin, He Qihua, Wang Xiaomin. Effect of Chinese herb Tripterygium wilfordii Hook F monomer triptolide on apoptosis of PC12 cells induced by A#beta#1-42. Acta Physiol Sin 2004; 56 (1): (in Chinese with English abstract).