牛膝多肽对体外培养的MPP^+诱导的大鼠多巴胺能神经元凋亡的保护作用  

Protective effects of achyranthes bidentata polypeptides(ABPP) against MPP^+ induced cell apoptosis in vitro rat dopaminergic neurons

作　　者：管菁菁[1] 王军[1] 顾晓松[2] 孙诚[2] 

机构地区：[1]南通大学附属医院老年医学科,南通226001 [2]南通大学神经再生重点实验室,南通226001

出　　处：《神经解剖学杂志》2016年第5期607-612,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31271260);教育部博士点基金(20110091120029);南通大学研究生科技创新计划项目(YKS14024)

摘　　要：目的:观察牛膝多肽(achyranthes bidentatapolypeptides,ABPP)对MPP^+(1-甲基-4-苯基-吡啶离子)损伤多巴胺能神经元的保护作用。方法:原代培养大鼠胚胎中脑多巴胺能神经元,与浓度50 ng/ml的ABPP或50ng/ml神经生长因子(NGF)共同孵育6 h,加入20μmol/L MPP^+损伤多巴胺能神经元12 h。使用四甲基偶氮唑盐(methyl thiaaolyl terazolium salt color imetry,MTT)比色法测定细胞活力;采用末端标记技术(Td T-mediated d UTP Nick-End Labeling,TUNEL)检测细胞凋亡;流式细胞仪检测细胞凋亡;Western Blot检测细胞凋亡相关蛋白Bcl-2和Bax的表达水平。结果:ABPP或NGF预处理6 h可增强多巴胺能神经元的细胞活力,抑制MPP^+损伤后多巴胺能神经元的凋亡,上调Bcl-2/Bax蛋白的表达水平。ABPP对大鼠多巴胺能神经元的保护作用优于NGF。结论:与NGF相比,ABPP发挥了更好的拮抗MPP^+对多巴胺能神经元损伤的作用,其机制可能与上调Bcl-2/Bax蛋白的表达水平相关。Objective： To investigate the protective effects of achyranthes bidentata polypeptides（ ABPP） on MPP~＋induced cell injury in cultured dopaminergic neurons. Methods： Cultured primary rat embryonic mesencephalic dopaminergic neurons were incubated with ABPP（ 50 ng / ml） or NGF（ 50 ng / ml） for 6 h,and then 20 μmol / L MPP~＋was added for 12 h to injury cells. Cell viability was assessed by methyl thiazolyl terazolium salt colorimetry assay（ MTT）. Cell apoptosis was evaluated by end labeling technique（ Td T mediated d UTP Nick End Labeling,TUNEL） and flow cytometry. Protein level of Bcl-2 and Bax were assayed by Western Blot. Results： 6 h-pretreatment with ABPP or NGF increased the viability of dopaminergic neurons,attenuated MPP~＋-induced cell apoptosis; up-regulated the expression levels of Bcl-2 / Bax protein. Moreover,the neuroprotective effects of ABPP were superior to those of NGF. Conclusion：ABPP has more potent neuroprotective effects as compared to NGF,which might be due to the up-regulation of the expression level of Bcl-2 / Bax induced by ABPP.

关 键 词：牛膝多肽 中脑多巴胺能神经元 MPP+ 细胞凋亡 神经保护 帕金森病 细胞培养 大鼠 

分 类 号：R742.5[医药卫生—神经病学与精神病学]