Abstract：Parkinson's disease (PD), the second most common neurodegenerative disease. It includes the major motor symptoms of akinesia, rigidity and tremor. A lot of study indicates that apoptosis is involved in the pathogenesis of PD. The present study is to investigate the mRNA expression of tumor suppressor gene P53, cysteinyl aspartate-specific proteinase-3 (Caspase-3), B cell lymphoma-2 (Bcl-2) and BCL-2-associated X protein (Bax) in peripheral blood before and after treatment in monkey (Macaca mulatta) with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Six healthy young rhesus monkeys were respectively daily injected a small dose (0.2 mg/(kg·d)) of MPTP. The behavioral manifestations of all monkeys were evaluated before and after the injection for 30 min, respectively. The forearm venous blood of rhesus monkeys were took at 0 week(non-injected MPTP) and 4, 8, 12, 16, 20, 24 week after injection MPTP. qRT-PCR detected P53, Bax, Bcl-2 and Caspase-3 mRNA expression in peripheral blood. The correlations between mRNA expression of P53, Caspase-3, Bcl-2 and Bax and the severity of behavior scores were analyzed. Behavioral observation indicated that from the 4 week, the experimental animals showed moderate bradykinesia, and then the symptoms gradually worsened, exercise capacity continued to decline, experimental animals showed a significant reduction in activity at 12 week, posture stiff and severe movement obstacle. After the 13 week of clinical symptoms was basically stable, the experimental animals showed slow movement, abnormal posture, muscle rigidity and resting tremor. qRT-PCR detected of apoptosis regulatory factors, compared with the 0 week, the expression of P53 and Caspase-3 significantly increased at the 4, 8, 12 week (P＜0.05), but the difference was not significantly at the 16, 20, 24 week; compared with the 0 week, Bcl-2/Bax ratio at the 4, 8, 12 week significantly decreased (P＜0.05), and the difference was not significantly at 16, 20 and 24 week. The mRNA expression of P53 and Caspase-3 were positively correlated with behavior scores in 0~12 week (P＜0.05). The above results suggested that when P53 and Casapse-3 significantly increased, the clinical symptoms of experimental animals also continued to intensify with the same period. The mRNA expression of P53 and Caspae-3 in peripheral blood can be used as one of the indicators for diagnosing PD, assessing the severity of PD.
万金城，涂文斌，徐芙蓉，等. 2009. MPTP慢性帕金森病小鼠纹状体差异蛋白质的研究[J].中国实验动物学报,17(4)：261-265.(Wan J C, Tu W B, Xu F R, et al . 2009. Proteomic analysis of striatum in the MPTP-induced mouse model of Parkinson's disease. Acta Laboratorium Animalis Scientia Sinica , 17(4): 261-265. )Bekris L M, Mata I F, Zabetian C P, et al. 2010. The genetics of Parkinson's disease[J]. Journal of Geriatric Psychiatry and Neurology ,23(4):228-242.Bezard E, Imbert C, Gross C E, et al. 1998. Experimental models of Parkinson's disease: from the static to the dynamic[J]. Reviews in the Neurosciences,9(2):71-90.Blandini F, Sinforiani E, Pacchett C, et al. 2006. Peripheral proteasome and caspase activity in Parkinson disease and Alzheimer disease[J]. Neurology,66:529-534.Burns R S, Chiueh C C, Markey S P, et al. 1983. A primate model of Parkinsonism: selective destruction of dopaminergic neurons in the pars compacts of the substantia nigra by MPTP[J]. Proceeding of the National Academy Sciences of the United States of America, 80(14):4546-4550.Calopa M, Bas J, Callen A, et al. 2010. Apoptosis of peripheral blood lymphocytes in Parkinson patients[J]. Neurobiology of Disease,38:1-7.Carvey P M, Punati A, Newman M B, et al. 2006. Progressive dopamine neuron loss in Parkinson's disease: the multiple hit hypothesis[J]. Cell Transplant,15(3): 239-250.Duchen M R. 2004. Mitochondria in health and disease: perspectives on a new mitochondrial biology[J]. Molecular Aspects of Medicine,25(4): 365-451.Eslamboli A. 2005. Marmoset monkey models of Parkinson's disease: Which model, when and why[J]? Brain Research Bulletin ,68: 140-149.Henry B, Fox S H, Crossman A R, et al. 2001. μ-and δ-opioid receptor antagonists reduce levodopa-induced dyskinesia in the MPTP-lesioned primate model of Parkinson's disease[J]. Experimental Neurology,171(1):139-146.Liu S M, Li X Z, Huo Y, et al. 2012. Protective effect of extract of Acanthopanax senticosus harms on dopaminergic neurons in Parkinson's disease mice[J]. Phytomedicine,19(7):631-638.Lofrumento D D，La P G，Abbrescia D I. et al. 2012. Valinomycin induced energy dependent mitochondrial swelling，cytochrome c release，cytosolic NADH/cytochrome c oxidation and apoptosis[J]. Apoptosis,16:1004-1013．Ly J, Grubb D, Lawen A, et al. 2003. The mitochondrial membrane potential in apoptosis; an update[J]. Apoptosis,8(2):115-128.Mogi M，Togari A，Kondo T，et a1. 2000. Nagatsu，Caspase activities and tumor necrosis factor receptor R1(p55)level ale elevated in the substantianigra from parkinsonian brain[J]. J Neural Transm,107:4-335.Muftuoglu, Elibol B, Ercan A, et al. 2003. Mitochondrial complex I and IV activities in leukocytes from patients with parkin mutations[J]. Mov. Disord,19:544-579.Petrozzi L, Nesti C, Orsucci D, et al. 2001. Cytogenetic analysis oxidative damage in lymphocytes of Parkinson's disease patients[J]. Neurol. Sci,22:83-84.Prigione A, Bengni B, Galbussera A, et al. 2006. Oxidative stress in peripheral blood mononuclear cells from patients with Parkinson's disease: negative correlation with levodopa dosage[J]. Neurobiol. Dis,23:36-43. Snow N A, Vingerhoets F J, Langston J W, et al. 2000. Pattern of dominergic loss in the striatum of humans with MPTP of humans with MPTP induced Parkinsonism[J]. J Neurol Neurosurg Pasychiatry,68(3):313-316.Tansey M G, Goldberg M S. 2010. Neuroinflammation in Parkinson's disease: its role in neuronal death and implications for therapeutic intervention[J]. Neurobiology Disease, 37(3):510-518.Toulouse A, Sullivan A M. 2008. Progress in Parkinson's disease-where do we stand[J]. Progress in Neurobiology,85(4):376-392. Twiddy D, Brown D G, Adrain C, et al. 2004. Pro-apoptotic proteins released from the mitochondria regulate the protein composition and caspase- Processing a ctivity of the native Apaf-1/caspase-9 apoptosome complex[J]. Journal of Biological chemistry, 279(19):19665-19682.Van Der Stelt M, Fox S H, Hill M, et al. 2005. A role for endocannabinoids in the generation of parkinsonism and levodopa-induced dyskinesia in MPTP-lesioned non-human primate models of Parkinson's disease[J]. The FASEB journal,19 (9):1140-1142.Xu C L, Wang Q Z, Sun L M, et a1. 2012. Asiaticoside: Attenuation of neurotoxicity induced by MPTP in a rat model of Parkinsonism via maintaining redox balance and up-regulating the ratio of Bcl-2/Bax[J]. Pharmacology, Biochemistry and Behavior ,100:413–418.Yamada M, Kida K, Amutuhaire W, et a1. 2010. Gene disruption of caspase-3 prevents MPTP-induced Parkinson’s disease in mice[J]．Bioehem Biophys Res Commun,402(2): 8-312．Zimin A V, Cornish A S, Maudhoo M D, et al. 2014. A new rhesus macaque assembly and annotation for next-generation sequencing analyses[J]. Biol. Direct,9 (1):20.
