1-三氯甲基-l,2,3,4-四氢化-β-咔啉(TaClo)介导的体外培养多巴胺能神经元凋亡及其机制  

Dopaminergic neuron apoptosis induced by 1-trichloromethyl-1,2,3,4tetrahydro-carboline(TaClo) in vitro and its mechanism

作　　者：杜鑫瑞[1] 辛涛[1] 滕良珠[1] 邴国英[2] 庞琦[1] 

机构地区：[1]山东大学附属省立医院神经外科,济南250021 [2]肯塔基大学解剖与神经生物学系,美国肯塔基州列克星敦市40536

出　　处：《山东大学学报（医学版）》2011年第8期41-44,83,共5页Journal of Shandong University:Health Sciences

摘　　要：目的研究TaC lo介导体外培养多巴胺能神经元细胞凋亡及其机制。方法分别用不同浓度的TaC lo(50、100、200、500、1 000μm o l/L)对原代神经元-胶质细胞混合培养和原代神经元培养的细胞进行干预,作用72h,细胞酪氨酸羟化酶(tyrosine hydroxy lase,TH)免疫细胞化学染色后多巴胺神经元计数;分别用浓度为50和200μm o l/L的TaC lo干预混合培养的细胞,作用1、24h后,W estern-b lot法测Caspase-3蛋白表达。结果 TaC lo作用后多巴胺能神经元数量明显下降,50μm o l/L浓度的TaC lo诱发40%的多巴胺能神经元死亡;当TaC lo浓度为200μm o l/L时,接近90%的多巴胺能神经元死亡(P<0.01、P<0.001);TaC lo(50、100、200μm o l/L)诱导的多巴胺能神经元死亡具有明显的剂量依赖性,结果有统计学意义(P<0.001)。W estern-b lot显示,TaC lo作用2个时间点,Caspase-3表达增高,表达均具有药物剂量依赖性。结论 TaC lo可以导致多巴胺能神经元凋亡,这一过程是通过活化的caspase-3蛋白表达等通路完成的。Objective To study the toxic effect and mechanisms of TaClo on rat dopaminergic neurons. Methods Pri- mary neuron-enriched and mesencephalic cultures were exposed to five different doses of TaClo （50, 100, 200, 500, and 1 000 ixmol/L） for 72 hours. Potential neurotoxicity was detected in morphology and by TH staining. Caspase-3 was detected by Western blot after exposure to two different doses of TaClo （50 and 200 ixmol/L） for 1 hour and 24 hours. Results TaClo induced dopaminergic neuronal loss. TaClo induced 40% of neuronal loss at 50 txmol/L, and a- bout 90% at 200 ixmol/L（ P 〈 0. 01, P 〈 0.001 ）. Under the same experimental condition, TaClo induced dopaminergic neuronal loss in a dose-dependent manner from 50 ixmol/L to 200 ixmol/L（ P 〈 0. 001 ）. Within one hour and twenty- four hours challenged with TaClo, cleaved caspase-3 was increased in both TaClo groups （50 and 200 txmol/L）, and activity of caspase-3 seemed to be increased in a dose-dependent manner. Conclusion TaClo induces dopaminergic neuronal apoptosis. The mechanism is that caspase-3 is activated by TaClo stimuli in dopaminergic neurons.

关 键 词：多巴胺能神经元 体外培养 TaClo Caspase.3 细胞凋亡 

分 类 号：R742.5[医药卫生—神经病学与精神病学]