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帕金森(Parkinson’s disease,PD)是一种以运动功能障碍为主要特征的神经退行性疾病,大量研究表明细胞凋亡参与了PD的发病机制。PD动物模型对于深入研究PD病因及发病机理、进行有效预防和提高治疗效果将起到重要作用。为探讨PD恒河猴(Macaca mulatta)外周血中肿瘤抑制基因P53、B细胞淋巴瘤-2(B cell lymphoma-2,Bcl-2)、B细胞淋巴瘤-2相关X蛋白(BCL-2-associated X protein,Bax)和半胱天冬氨酸蛋白酶-3(cysteinyl aspartate-specific proteinase-3,Caspase-3)mRNA表达水平与行为学变化之间的关系,本研究选取健康青年恒河猴6只,以0.2 mg/(kg·d)小剂量、多次重复肌内注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methy-4-pheny-l,2,3,6-tetrahydropyridine,MPTP)建立了PD恒河猴模型。于每次MPTP给药前后观察记录动物行为学表现30 min。分别于未注射MPTP(第0周),注射MPTP后第4、8、12、16、20和24周采集恒河猴前臂静脉血,采用qRT-PCR检测外周血P53、Bax、Bcl-2和Caspase-3 mRNA表达水平。行为学观察结果显示,第4周时,实验动物表现出温和的运动迟缓,随后症状逐渐加重,运动能力持续下降,到12周时实验动物均表现出活动量显著减少,姿势僵硬和严重的运动障碍。13周后临床症状基本稳定,实验动物表现出运动迟缓,姿势异常,肌强直和静止性震颤等。qRT-PCR结果显示,与0周相比,注射MPTP后第4、8、12周P53和Caspase-3 mRNA表达显著升高(P<0.05),而第16、20、24周差异不显著;与0周相比,Bcl-2/Bax比值在第4、8、12周显著降低(P<0.05),而第16、20、24周差异不显著。0~12周行为学评分与外周血P53和Caspase-3 mRNA表达水平呈正相关(r=0.975,r=0.956;P<0.05)。结果表明,当外周血P53和Caspase-3 mRNA表达水平显著升高时临床症状也处于逐渐加重的阶段,因此通过检测血液中凋亡调控基因的表达,能反应出PD恒河猴的疾病进程。研究结果可为PD诊断和疾病进程监测提供辅助参考。
Parkinson’s disease (PD) is a neurodegenerative disease characterized by motor dysfunction. A large number of studies have shown that apoptosis is involved in the pathogenesis of PD. PD animal model for further study of etiology and pathogenesis of PD, to effectively prevent and improve the therapeutic effect will play an important role. To investigate the expression of tumor suppressor gene P53, B cell lymphoma-2 (Bcl-2) and B-cell lymphoma-2 related protein B (BCL-2- associated X protein (Bax) and caspase-3 (Caspase-3) mRNA expression and behavioral changes in this study selected healthy young Rhesus 6 Only with a small dose of 0.2 mg / (kg · d), repeated intramuscular injection of 1-methy-4-pheny- l, 2,3,6-tetrahydropyridine, MPTP). Animals were observed for behavioral performance 30 min before and after each MPTP administration. Rhesus monkey forearm venous blood was collected at 4, 8, 12, 16, 20 and 24 weeks after MPTP injection (week 0), respectively. The levels of P53, Bax and Bcl-2 in peripheral blood were detected by qRT- Caspase-3 mRNA expression level. Behavioral observations showed that, at week 4, the experimental animals showed mild slowness of movement, followed by a gradual increase in symptoms and a continued decline in their ability to exercise. At 12 weeks, animals showed significantly reduced activity, stiff posture, and severe exercise obstacle. After 13 weeks, the clinical symptoms were basically stable. The experimental animals showed sluggishness, abnormal posture, muscle rigidity and rest tremor. qRT-PCR results showed that compared with 0 week, the expression of P53 and Caspase-3 mRNA at 4, 8 and 12 weeks after MPTP injection were significantly increased (P <0.05), while there was no significant difference at 16th, Compared with 0 week, the Bcl-2 / Bax ratio decreased significantly at the 4th, 8th and 12th week (P <0.05), while the difference was not significant at the 16th, 20th and 24th weeks. The behavioral score at 0-12 weeks was positively correlated with the expression of P53 and Caspase-3 mRNA in peripheral blood (r = 0.975, r = 0.956; P <0.05). The results showed that when the expression of P53 and Caspase-3 mRNA in peripheral blood increased significantly, the clinical symptoms were also gradually aggravated. Therefore, by detecting the expression of apoptosis-regulating genes in blood, the course of disease of PD rhesus could be reflected. The results of the study can provide a supplementary reference for the diagnosis of PD and the monitoring of disease progression.