帕金森病食蟹猴系统性模型构建与在体临床过程的动态评价研究
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摘要
目的应用行为学与分子影像学的方法在体动态评价帕金森病食蟹猴系统性模型的临床变化特征,为药物、干细胞等治疗的临床前研究提供稳定有效的PD灵长类动物模型。方法 7只10～15岁的健康食蟹猴连续静脉注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)0.2 mg/kg体质量,诱导系统性PD模型,持续观察PD症状进展3个月,随后给予左旋多巴干预。应用PD评分量表评价动物的临床症状严重程度,用EthoVision动物运动轨迹跟踪系统分析随意运动距离、运动轨迹的变化,用正电子发射断层显像技术(Positron emission tomography,PET)分子显像剂~(18)F-AV-133评估纹状体多巴胺能神经元的功能状态。结果所有动物在给予MPTP注射14 d后出现了典型的包括震颤、肌僵直、运动迟缓等PD症状,1个月临床评分达到峰值(21.43±5.35)。随后PD症状趋于稳定,连续观察至2个月(18.43±3.87)、3个月(18.14±3.53),与MPTP注射结束时(14.43±1.90)相比,临床评分均显著升高(P<0.05)。在3个月随意运动距离(809.77±401.15)cm较基线(8627.46±5751.04)cm显著降低(P<0.01)。~(18)F-AV-133双侧纹状体平均特异性摄取率(Sur)在3个月时(0.16±0.03)较基线(1.66±0.58)显著降低(P<0.01)。在给予L-Dopa干预后可见PD症状显著改善,临床评分(12.86±3.63)较之模型期显著降低(P<0.05),而随意运动距离则显著增加(P<0.05)。结论本研究构建的PD食蟹猴系统性模型临床症状持续稳定,纹状体多巴胺能神经损害,对L-Dopa干预有效,全程无自发性恢复,更近似地在体动态模拟了PD的临床特征,以期为PD未来研究提供实验依据。
Objective To model the systemic parkinsonism and in vivo dynamically evaluate the clinical characteristics by using behavioral and molecular imaging method in cynomolgus monkeys, and provide a reliable Parkinson's disease(PD)model for preclinical studies of drug and stem cell and translational research. Method Seven healthy middle aged cynomolgus monkeys with age between 10-15 years were intravenously injected of MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)(0.2 mg/kg)to induce a systemic PD model, continuously observe the evolution of clinical symptoms up to 3 months followed by L-Dopa intervention. Behavior analysis including rating scale for evaluating the severity of clinical symptoms, the EthoVision tracking system for measuring the moved distance, motion trajectory, ~(18)F-AV-133 the Positron emission tomography(PET)imaging for detecting the functional integrity of striatal dopaminergic system. Result All animals showed the typical PD symptoms including resting tremor, rigidity, and bradykinesia etc. at 14 days or so when MPTP injection was stopped. The clinical score reached peak at 1 month(21.43±5.35), and then the symptoms remain stabilized and continued. The clinical scores were(14.43±1.90) at the end of MPTP injection,(18.43±3.87) in the 2 months and(18.14±3.53) in 3 months post MPTP respectively. The moved distance was significantly lower at 3 months(809.77±401.15)cm than the baseline(8627.46±5751.04)cm(P<0.01). The Specific uptake rate(Sur)of the ~(18)F-AV-133 was significantly reduced at 3 months(0.16±0.03)than the baseline(1.66±0.58)(P<0.01). PD symptoms were significantly improved after L-Dopa intervention, the clinical score(12.86±3.63)was significantly decreased(P<0.05)while the moved distance was significantly increased(P<0.05)than that before. Conclusion A systemic PD is modeled with continuously stable clinical symptoms with no spontaneous recovery, and the damage of functional integrity of striatal dopaminergic system, and the positive response to L-Dopa intervention, which closely simulates the clinical features of PD and provides a stable and reliable PD model for preclinical translational studies.
Objective To model the systemic parkinsonism and in vivo dynamically evaluate the clinical characteristics by using behavioral and molecular imaging method in cynomolgus monkeys, and provide a reliable Parkinson's disease(PD)model for preclinical studies of drug and stem cell and translational research. Method Seven healthy middle aged cynomolgus monkeys with age between 10-15 years were intravenously injected of MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)(0.2 mg/kg)to induce a systemic PD model, continuously observe the evolution of clinical symptoms up to 3 months followed by L-Dopa intervention. Behavior analysis including rating scale for evaluating the severity of clinical symptoms, the EthoVision tracking system for measuring the moved distance, motion trajectory, ~(18)F-AV-133 the Positron emission tomography(PET)imaging for detecting the functional integrity of striatal dopaminergic system. Result All animals showed the typical PD symptoms including resting tremor, rigidity, and bradykinesia etc. at 14 days or so when MPTP injection was stopped. The clinical score reached peak at 1 month(21.43±5.35), and then the symptoms remain stabilized and continued. The clinical scores were(14.43±1.90) at the end of MPTP injection,(18.43±3.87) in the 2 months and(18.14±3.53) in 3 months post MPTP respectively. The moved distance was significantly lower at 3 months(809.77±401.15)cm than the baseline(8627.46±5751.04)cm(P<0.01). The Specific uptake rate(Sur)of the ~(18)F-AV-133 was significantly reduced at 3 months(0.16±0.03)than the baseline(1.66±0.58)(P<0.01). PD symptoms were significantly improved after L-Dopa intervention, the clinical score(12.86±3.63)was significantly decreased(P<0.05)while the moved distance was significantly increased(P<0.05)than that before. Conclusion A systemic PD is modeled with continuously stable clinical symptoms with no spontaneous recovery, and the damage of functional integrity of striatal dopaminergic system, and the positive response to L-Dopa intervention, which closely simulates the clinical features of PD and provides a stable and reliable PD model for preclinical translational studies.
引文
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[22] Chen M K,Kuwabara H,Zhou Y,et al.VMAT2 and dopamine neuron loss in a primate model of Parkinson's disease [J].Journal of neurochemistry,2008,105(1):78-90.
[23] Kostrzewa R M,Nowak P,Kostrzewa J P,et al.Peculiarities of L:-DOPA treatment of Parkinson's disease [J].Amino acids,2005,28(2):157-164.
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[25] Stewart A M,Grieco F,Tegelenbosch Ra J,et al.A novel 3D method of locomotor analysis in adult zebrafish:Implications for automated detection of CNS drug-evoked phenotypes [J].Journal of neuroscience methods,2015,255:66-74.
[26] Lee K N,Pellom S T,Oliver E,et al.Characterization of the guinea pig animal model and subsequent comparison of the behavioral effects of selective dopaminergic drugs and methamphetamine [J].Synapse,2014,68(5):221-233.
[27] Walton A,Branham A,Gash D M,et al.Automated video analysis of age-related motor deficits in monkeys using EthoVision [J].Neurobiology of aging,2006,27(10):1477-1483.
[28] Liu N,Yue F,Tang W P,et al.An objective measurement of locomotion behavior for hemiparkinsonian cynomolgus monkeys [J].Journal of neuroscience methods,2009,183(2):188-194.
[2] Blaszczyk J W.Nigrostriatal interaction in the aging brain:new therapeutic target for Parkinson's disease [J].Acta neurobiologiae experimentalis,2017,77(1):106-112.
[3] Choudhury G R,Daadi M M.Charting the onset of Parkinson-like motor and non-motor symptoms in nonhuman primate model of Parkinson's disease [J].PloS one,2018,13(8):e0202770.
[4] Alexander P K,Lie Y,Jones G,et al.Management Impact of Imaging Brain Vesicular Monoamine Transporter Type 2 in Clinically Uncertain Parkinsonian Syndrome with (18)F-AV133 and PET [J].Journal of nuclear medicine:official publication,Society of Nuclear Medicine,2017,58(11):1815-1820.
[5] 张娜,窦德强.帕金森病分子病理机制研究进展 [J].神经药理学报,2013,3(2):35-42.
[6] Gerlach M,Riederer P,Przuntek H,et al.MPTP mechanisms of neurotoxicity and their implications for Parkinson's disease [J].European journal of pharmacology,1991,208(4):273-286.
[7] Gubellini P,Kachidian P.Animal models of Parkinson's disease:An updated overview [J].Revue neurologique,2015,171(11):750-761.
[8] Jackson-Lewis V,Przedborski S.Protocol for the MPTP mouse model of Parkinson's disease [J].Nature protocols,2007,2(1):141-151.
[9] Katzenschlager R,Lees A J.Treatment of Parkinson's disease:levodopa as the first choice [J].Journal of neurology,2002,249(2):ii19-ii24.
[10] Kurlan R,Kim M H,Gash D M.Oral levodopa dose-response study in MPTP-induced hemiparkinsonian monkeys:assessment with a new rating scale for monkey parkinsonism [J].Movement disorders:official journal of the Movement Disorder Society,1991,6(2):111-118.
[11] Rose C,De Heer R C,Korte S,et al.Quantified tracking and monitoring of diazepam treated socially housed cynomolgus monkeys [J].Regulatory toxicology and pharmacology:RTP,2012,62(2):292-301.
[12] Ingram E M,Augustin J,Ellis M D,et al.Evaluating sub-lethal effects of orchard-applied pyrethroids using video-tracking software to quantify honey bee behaviors [J].Chemosphere,2015,135:272-277.
[13] Liu Y,Yue F,Tang R,et al.Progressive loss of striatal dopamine terminals in MPTP-induced acute parkinsonism in cynomolgus monkeys using vesicular monoamine transporter type 2 PET imaging ([(18)F]AV-133) [J].Neuroscience bulletin,2014,30(3):409-416.
[14] Lane M A.Nonhuman primate models in biogerontology [J].Experimental gerontology,2000,35(5):533-541.
[15] Emborg M E.Nonhuman Primate Models of Neurodegenerative Disorders [J].ILAR journal,2017,58(2):190-201.
[16] 江鹏亮,汤球,余琛琳,等.非人灵长类动物模型在医学研究中的应用概况 [J].实验动物科学,2010,27(6):59-64.
[17] Potts L F,Wu H,Singh A,et al.Modeling Parkinson's disease in monkeys for translational studies,a critical analysis [J].Experimental neurology,2014,256(6):133-143.
[18] Fox S H,Brotchie J M.The MPTP-lesioned non-human primate models of Parkinson's disease.Past,present,and future [J].Progress in brain research,2010,184:133-157.
[19] Emborg M E,Zhang Z,Joers V,et al.Intracerebral transplantation of differentiated human embryonic stem cells to hemiparkinsonian monkeys [J].Cell transplantation,2013,22(5):831-838.
[20] Bezard E,Dovero S,Prunier C,et al.Relationship between the appearance of symptoms and the level of nigrostriatal degeneration in a progressive 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned macaque model of Parkinson's disease [J].The Journal of neuroscience:the official journal of the Society for Neuroscience,2001,21(17):6853-6861.
[21] Masilamoni G J,Smith Y.Chronic MPTP administration regimen in monkeys:a model of dopaminergic and non-dopaminergic cell loss in Parkinson's disease [J].Journal of neural transmission,2018,125(3):337-363.
[22] Chen M K,Kuwabara H,Zhou Y,et al.VMAT2 and dopamine neuron loss in a primate model of Parkinson's disease [J].Journal of neurochemistry,2008,105(1):78-90.
[23] Kostrzewa R M,Nowak P,Kostrzewa J P,et al.Peculiarities of L:-DOPA treatment of Parkinson's disease [J].Amino acids,2005,28(2):157-164.
[24] Schneider J S,Pioli E Y,Jianzhong Y,et al.Levodopa improves motor deficits but can further disrupt cognition in a macaque Parkinson model [J].Movement disorders:official journal of the Movement Disorder Society,2013,28(5):663-667.
[25] Stewart A M,Grieco F,Tegelenbosch Ra J,et al.A novel 3D method of locomotor analysis in adult zebrafish:Implications for automated detection of CNS drug-evoked phenotypes [J].Journal of neuroscience methods,2015,255:66-74.
[26] Lee K N,Pellom S T,Oliver E,et al.Characterization of the guinea pig animal model and subsequent comparison of the behavioral effects of selective dopaminergic drugs and methamphetamine [J].Synapse,2014,68(5):221-233.
[27] Walton A,Branham A,Gash D M,et al.Automated video analysis of age-related motor deficits in monkeys using EthoVision [J].Neurobiology of aging,2006,27(10):1477-1483.
[28] Liu N,Yue F,Tang W P,et al.An objective measurement of locomotion behavior for hemiparkinsonian cynomolgus monkeys [J].Journal of neuroscience methods,2009,183(2):188-194.