改良6-OHDA注射法建立早期帕金森病大鼠模型The Establishment of Rat Model of Early Stage Parkinson's Disease by Improved Method with 6-OHDA Injection
韩飞,李成朋,魏操,杨梅,周波,张春林,焦玲,官志忠,禹文峰
HAN Fei,LI Chengpeng,WEI Cao,Yang Mei,ZHOU Bo,ZHANG Chunlin,JIAO Ling,GUAN Zhizhong,YU Wenfeng
摘要(Abstract):
目的:改良6-羟基多巴胺(6-OHDA)立体定位注射法建立早期帕金森病(PD)大鼠模型。方法:40只雄性SD大鼠随机分为control组10只,model组30只;于手术前30 min,腹腔注射地昔帕明(25 mg/kg)及帕吉林(5 mg/kg),control组采用立体定位右侧纹状体注射生理盐水,model组注射6-OHDA;术后第3、4周采用圆筒测试及旋转测试测定大鼠的左前肢使用率和旋转速度,免疫组化方法检测黑质内酪氨酸羟化酶(TH)阳性神经元数量。结果:圆筒测试显示madel组大鼠左前肢使用率低于control组(P<0.01),旋转测试显示model组平均向左侧旋转速度(3.48圈/min),明显快于对照组(P<0.01),PD模型成功率为89.66%;免疫组化显示,model组右侧中脑黑质致密部(SNc)部位TH阳性神经元数目较control组右侧减少(44.81±2.89)%,差异有统计学意义(P<0.01),符合临床早期PD诊断标准。结论:改良的6-OHDA注射法建立早期PD大鼠模型简单易行,成功率高。
Objective:To establish rat model of early stage of Parkinson's disease( PD) with improved method of stereotactic injection of 6-OHDA. Methods:Forty male SD rats were randomly divided into control group( group C,10 rats) and model group( group PD,30 rats). Desipramine( 25mg/kg) and pargyline( 5 mg/kg) were injected intraperitoneally 30 minutes before surgery,followed by stereotactic injection of saline( control group) or 6-OHDA( model group) in striatum. Cylinder test and rotation experiments were adopted to evaluate the behavior in 3 and 4 weeks after surgery. Immunohistochemical staining was adopted to analyze the positive cells of tyrosine hydroxylase( TH) in substantia nigra compacta( SNc). Results:Cylinder test showed that the utilization rate of the left forelimb in group PD was lower than that in group C( P < 0. 01). Rotation experiments showed that the average number of turns in group PD was about 3. 48 turns/min while there was no rotation in group C( P < 0. 01). The success rate of PD model was 89. 66%. Immunohistochemistry showed TH positive neurons in right SNc of PD model decreased about 44. 81% compared with that of group C,which was in accordance with the criteria of early stage of PD. Conclusions:The improved method of 6-OHDA injection is a simple,feasible and highly successful approach to establish the model of early PD.
关键词(KeyWords):
6-羟基多巴胺;帕金森病;大鼠;多巴胺神经元;酪氨酸羟化酶
6-Hydroxydopamine;Parkinson's disease;rats;dopamine neurons;tyrosine hydroxylase
基金项目(Foundation): 国家自然科学基金(81360199);; 教育部科学技术研究项目(213032A);; 贵州省国际科技合作计划项目[批准号:黔科合外G字(2013)7026号];; 贵州省创新计划项目[黔教合协同创新中心(2014)06];; 贵州省教育厅项目(2015年贵州省普通高等学校地方病和少数民族疾病防控创新团队);; 贵州省科技计划课题任务书[黔科合重大专项字(2014)6008]
作者(Author):
韩飞,李成朋,魏操,杨梅,周波,张春林,焦玲,官志忠,禹文峰
HAN Fei,LI Chengpeng,WEI Cao,Yang Mei,ZHOU Bo,ZHANG Chunlin,JIAO Ling,GUAN Zhizhong,YU Wenfeng
DOI: 10.19367/j.cnki.1000-2707.2017.03.003
参考文献(References):
- [1]Dauer W,Przedborski S.Parkinson's disease:mechanisms and models.[J].Neuron,2003(6):889-909.
- [2]孙倩,陈伟,陈生弟.帕金森病早期诊断的生物学标志物[J].中国现代神经疾病志,2013(8):667-672.
- [3]Kirik D,Rosenblad C,Bj9rklund A.Characterization of behavioral and neurodegenerative changes following partial lesions of the nigrostriatal dopamine system induced by intrastriatal 6-hydroxydopamine in the rat.[J].Experimental Neurology,1998(2):259-277.
- [4]Segura-Aguilar J,Paris I,Muoz P.The need of a new and more physiological preclinical model for Parkinson’s disease[J].Cellular and Molecular Life Sciences,2016(7):1381-1382.
- [5]Jagmag SA,Tripathi N,Shukla SD,et al.Evaluation of models of parkinson's disease[J].Frontiers in Neuroscience,2016(92):503.
- [6]Paxinos G,Watson CR.The rat brain in stereotaxic coordinates[M].New York:Academic Press.1986:6
- [7]Decressac M,Mattsson B,Bj9rklund A.Comparison of the behavioural and histological characteristics of the 6-OHDA andα-synuclein rat models of Parkinson's disease[J].Experimental Neurology,2012(1):306-315.
- [8]Elbaz A,Carcaillon L,Kab S,et al.Epidemiology of Parkinson's disease.[J].Revue Neurologique,2015(1-2):129-151.
- [9]Grealish S,Mattsson B,Draxler P,et al.Characterisation of behavioural and neurodegenerative changes induced by intranigral 6-hydroxydopamine lesions in a mouse model of Parkinson’s disease[J].European Journal of Neuroscience,2010(12):2266-2278.
- [10]李进,孙杰,赵楠,等.3种不同脑区定点注射6-OHDA单侧损伤大鼠帕金森病模型的比较研究[J].河北医科大学学报,2016(6):667-671.
- [11]Simola N,Morelli M,Carta AR.The 6-Hydroxydopamine model of parkinson’s disease[J].Neurotoxicity Research,2007(3-4):151-167.
- [12]Acton PD,Mozley PD.Single photon emission tomography imaging in parkinsonian disorders:a review.[J].Behavioural Neurology,2000(1-2):11-27.
- [13]Tobón-Velasco JC,Limón-Pacheco JH,OrozcoIbarra M,et al.6-OHDA-induced apoptosis and mitochondrial dysfunction are mediated by early modulation of intracellular signals and interaction of Nrf2 and NF-κB factors[J].Toxicology,2013(304):109-119.
- [14]Francardo V,Recchia A,Popovic N,et al.Impact of the lesion procedure on the profiles of motor impairment and molecular responsiveness to L-DOPA in the 6-hydroxydopamine mouse model of Parkinson's disease[J].Neurobiology of Disease,2011(3):327-340.
- [15]Ichitani Y,Okamura H,Nakahara D,et al.Biochemical and immunocytochemical changes induced by intrastriatal6-hydroxydopamine injection in the rat nigrostriatal dopamine neuron system:evidence for cell death in the substantianigra[J].Experimental Neurology,1994(2):269-278.
- [16]Timothy Schallert SMFJ.CNS plasticity and assessment of forelimb sensorimotor outcome in unilateral rat models of stroke,cortical ablation[J].Neuropharmacology,2000(5):777-787.
- [17]Henry B,Crossman AR,Brotchie JM.Characterization of a rodent model in which to investigate the molecular and cellular mechanisms underlying the pathophysiology of L-dopa-induced dyskinesia.[J].Advances in Neurology,1998(3):53-61.
- [18]Thiele SL,Warre R,Nash JE.Development of a Unilaterally-lesioned 6-OHDA Mouse Model of Parkinson's Disease[J].Journal of Visualized Experiments,2012(60):3234-3234.
- [19]王修敏.大鼠早期帕金森病模型的初建与评价[D].温州医学院,2011.
- [20]Truong L,Allbutt H,Kassiou M,et al.Developing a preclinical model of Parkinson's disease:A study of behaviour in rats with graded 6-OHDA lesions[J].Behavioural Brain Research,2006(1):1-9.
文章评论(Comment):
|
||||||||||||||||||
|
- 6-羟基多巴胺
- 帕金森病
- 大鼠
- 多巴胺神经元
- 酪氨酸羟化酶
6-Hydroxydopamine - Parkinson's disease
- rats
- dopamine neurons
- tyrosine hydroxylase
- 韩飞
- 李成朋
- 魏操
- 杨梅
- 周波
- 张春林
- 焦玲
- 官志忠
- 禹文峰
HAN Fei - LI Chengpeng
- WEI Cao
- Yang Mei
- ZHOU Bo
- ZHANG Chunlin
- JIAO Ling
- GUAN Zhizhong
- YU Wenfeng
- 韩飞
- 李成朋
- 魏操
- 杨梅
- 周波
- 张春林
- 焦玲
- 官志忠
- 禹文峰
HAN Fei - LI Chengpeng
- WEI Cao
- Yang Mei
- ZHOU Bo
- ZHANG Chunlin
- JIAO Ling
- GUAN Zhizhong
- YU Wenfeng