3-硝基丙酸预处理对多巴胺神经元生存预后影响的研究  

Experimental study on neuroprotective effects of preconditioning with 3NP on prognosis of dopaminergic neurons in substantia nigra of 6-OHDA-lesioned PD rats

作　　者：钟建新[1] 孙圣刚[2] 曹学兵[2] 钟森[3] 陈克强[1] 黄俊杰[1] 黎冠东[1] 

机构地区：[1]江门市中心医院神经内科,广东江门529071 [2]华中科技大学同济医学院协和医院神经内科,湖北武汉430030 [3]十堰市人民医院,湖北十堰44200

出　　处：《中国热带医学》2007年第3期343-346,共4页China Tropical Medicine

基　　金：国家自然科学基金(30170334);广东省医学科研基金(A2004724)资助项目

摘　　要：目的观察不同3-硝基丙酸(3-Nitropropionic acid,3NP)预处理方式对黑质多巴胺神经元(Dopaminergic neurons,DA)存活能力的影响,寻求3NP预处理保护黑质DA神经元的最佳预处理方式。方法以6-OHDA为损毁剂建立大鼠在体及离体PD模型。在体3NP预处理组于造模前2h、1d、3d、5d、7d分别腹腔注射不同剂量3NP(10mg/kg、20mg/kg、40mg/kg),观察6-OHDA损毁2周后大鼠的行为学;黑质脑片体系在含0.01mM6-OHDA的人工脑脊液Ⅱ(ACSFⅡ)中加入不同浓度3NP(0.1mM、0.3mM、0.6mM)共孵育脑片至不同时间点(0、5h、1h、2h、3h)。采用免疫组织化学方法检测黑质酪氨酸羟化酶(Tyrosine hydroxylase,TH)蛋白表达。结果20mg/kg3NP预处理的各个时间点均可以显著改善APO诱发PD大鼠的旋转行为;同时3NP可以抑制6-OHDA诱导的黑质TH样免疫反应阳性细胞数减少,其中在离体脑片体系中,0.3mM3NP预处理1h、2h时间差异具有显著性(P<0.05或P<0.01);在体试验中20mg/kg3NP预处理的各个时间点差异均有显著性(P<0.05),其中以3NP预处理3d显著性差异更加明显(P<0.01)。结论适宜剂量3NP预处理对PD大鼠黑质DA神经元具有保护效应,3NP预处理可能成为延缓DA神经元变性的新策略。Objective To study the effects of preconditioning pattern with 3 - nitroproplonic acid （3NP） on the survival of dopaminergic （DA） neurons in SN and the neuroprotection of 3NP preconditioning on the survival of DA neurons. Methods PD models were established with unilateral stereotactically microlnjection of 6 - Hydroxydopaminc（6- OHDA, 8μg/4μl） into the right side of SN adult male Sprague - Dawley （SD） rats, weighting 180 - 200g. The rats in 3NP preconditioned group（PC） were intraperitoneaily pretreated with different dosages of3NP （10,20,40mg/kg, respectively） and at different time （2 hours, 1,3,5,7 days, respectively） before making PD models. The rat SN slice models, which were suitable for investigating neurotuxicity on DA neurons, were prepared by incubating in artificial cerebrospinal fluid Ⅱ（ACSF Ⅱ） containing 6 - OHDA（0.01mM） for different duration（0.5, 1,2, 3beurs） . PC dices were incubated in ACSF II buffer solution containing 6 - OHDA （0.01mM） and 3NP（0.1, 0.3, 0.6 mM, respectively）. DA neurons within SN were labeled by immunohistochemistric staining for tyrosine hydroxylase （TH） coupled with diaminobenzidine. Apomorphine - induced rotational behaviors were measured 2 weeks after 6 - OHDA - lesion. Results The num- ber of TH - IR（immuno - reactive） - positive staining neurons in SN of 6 - OHDA lesioned side in PC group with 3NP at dose of 20mg/kg pretreated intraperitoneally at 2h,1d,3d and 5d ,especially the group with 20mg/kg 3NP 3d prior to the 6 - OHDA - lesioned, increased statistically than that in PD groups （ P 〈 0.05, P 〈 0.01 ）. Similarly, the number of TH - IR positive neurons in 0.3mM 3NP PC slice group incubated for lh and 2h were increased markedly than that in the 6 - OHDA SN slice group （ P 〈 0.01, P 〈 0.05, respectively） .Rats in PC group with 20mg/kg 3NP showed a significant improvement in APO - induced rotation behavior than 6 - OHDA - lesioned group, such as the elongation of rotational latency, the mean rota

关 键 词：3-硝基丙酸 预处理 多巴胺神经元 6-羟基多巴胺 酪氨酸羟化酶 神经元保护 

分 类 号：R741[医药卫生—神经病学与精神病学]