Establishment And Evaluation Of A Preclinic Stage Model Of Pakinson's Disease In C57bl/6j Mice Induced By MPTP

Objective:To further search for Parkinson's disease (parkinson disease, PD) diagnosis of pre-clinical indicators and to understand the development process of chronic Parkinson's disease,which we using mice to explore C57BL/6j early mouse model of Parkinson disease. We were observed mice behavior and morphological changes that to find the early diagnosis of Parkinson's disease biomarker.Methods:60C57BL/6j healthy adult male mice, which weighing 22-25g, were randomly divided into control group and model group. model: intraperitoneal injection of 1 - methyl -4-phenyl 1,2,3,6– tet-rahydropyridine , (MPTP, 4mg/kg, 0.9%sterile saline solution, the existing service is used). Model group was divided into four groups, each group successive administration of the morning 5,10,15,20 d, in the administration of the 1d, 3d,6d by traction test, swimming test in mice behavioral test. After 7days each with drawal, and attentive extraction blood perfusion in mice brain samples produced. Minimally invasive extraction of mouse plasma by high performance liquid chromatography 8 - hydroxy-deoxyguanosine (8-OHdG), glutathione (GSH) changes in uric acid was detected by biochemical testing (UA) values change. Morphological study mainly for anti-tyrosine hydroxylase (tyrosine hydroxylase, TH) immunohistochemistry, Nissl (Nissl) staining to observe the histological changes and substantia nigra dopamine (DA) neurons changes observed under light microscope the count, the shape and statistical analysis. Electron microscopy within the substantia nigra in mice ultrastructural changes.Results:Control group and model group: (1) behavior detection coordination of limb movements in mice in the control group and model group uncoordinated limb observation conditions were not found in animals, motion detection score statistics showed that the model group and control group by was no statistically significant difference (P> 0.05). (2)biochemical testing, HPLC results showed that the model group of 8-OHdG, GSH, UA levels with the delivery time are not the same as the changes occurred in the model group compared with the control group increased 8-OHdG is a the trend is to reduce the latter two changes, the model group compared with the control group with significant difference (P <0.05). (3) morphological changes, Nissl staining showed that the mice in the experimental model of substantia nigra in the brain scattered pyknosis of neurons, large neurons stained deep, normal control mice in the substantia nigra nerve argument structure is normal. TH immunohistochemistry showed that the control group, net Tissue nigra (SNc) TH-positive neurons intensive, the overall area, is distributed in the SNr; MPTP model group, the number of TH-positive neurons was significantly reduced, the cell processes to reduce change round, cytoplasm and fibers colored shallow, irregular cell distribution, SNr area fiber decreased. MPTP model mice with substantia nigra (SNC) region the number of TH-positive neurons compared with the control group mice were significantly different (P <0.05). (4) The ultrastructure of the substantia nigra in mice with pathological changes: normal control mice substantia nigra cells and rough endoplasmic reticulum (RER), polysomes and normal mitochondrial ultrastructure; administration 5,10 d super mice micro-structure is normal, and 15,20 d mice can be seen in the substantia nigra in the rough endoplasmic reticulum dilatation, reduced rough endoplasmic reticulum, ribosomes and mitochondria swelling depolymerization, ridge shorter, vacuolization so, these pathological changes with time and increased delivery.Conclusions:The present study, repeated administration of low dose MPTP long way to establish a mouse pre-clinical Parkinson's disease animal model. Ultrastructural morphology of the model changes, biochemical and behavioral testing and other research results can be used as a reference index of early Parkinson's disease, Parkinson's disease for the early screening, diagnosis and timely intervention to control the development of the disease reference value.