Survivin在帕金森病大鼠中对神经元凋亡的调控作用

Role of Survivin during the process of neuronal apoptosis in a rat model of Parkinson’s disease

摘要:

目的：研究生存素（Survivin）在大鼠帕金森病（Parkinson’s disease，PD）模型中对黑质神经元凋亡的调控作用。方法：成年健康雄性SD大鼠，随机分为4组，对照组（n=8）；PD模型组（n=8）；PD模型+腺病毒空载体组（n=8）；PD模型+Survivin治疗组（n=8）。造模成功1周后，对照组和PD模型组不予以处理，PD模型+腺病毒空载体组在大鼠纹状体的同一坐标点注射腺病毒空载体，PD模型+Survivin治疗组在大鼠纹状体的同一坐标点注射携带Survivin基因腺病毒，治疗结束后每周对大鼠颈部皮下注射阿扑吗啡（Apomorphine，APO）进行行为学评价，行为学上出现差异后收集各组大鼠脑组织标本。分别运用TUNEL法观察黑质神经元凋亡情况，qRT-PCR检测Caspase 3和Caspase 9的mRNA表达情况水平，Western blot检测Cleaved-Caspase 3和Cleaved-Caspase 9的蛋白水平，评价Survivin对神经元凋亡的调控作用。结果：治疗结束后第3周，PD模型+Survivin治疗组与同期PD模型组及PD模型+腺病毒空载体组比较，大鼠旋转圈数减少（F=22.315，P=0.000，P=0.000）；TUNEL结果显示，与对照组比较，PD模型组及PD模型+腺病毒空载体组大鼠脑黑质神经元细胞凋亡明显；与PD模型组及PD模型+腺病毒空载体组比较，PD模型+Survivin治疗组黑质神经元细胞凋亡程度明显减轻（F=38.912，P=0.000，P=0.000）；此外，qRT-PCR和Western blot的结果显示，与PD模型组及PD模型+腺病毒空载体组比较，Survivin能够显著抑制神经元细胞中Caspase 3/9在mRNA水平（F=338.429，P=0.000，P=0.000；F=283.352，P=0.000，P=0.000）和Cleaved-Caspase 3/9在蛋白水平（F=640.262，P=0.000，P=0.000；F=354.210，P=0.000，P=0.000）的表达。结论：Survivin能够通过抑制PD大鼠脑组织中Caspase 3和Caspase 9的表达，减少脑内黑质神经元的凋亡，进而达到对脑黑质退行性病变的治疗作用，减轻PD相关症状

Abstract:

Objective：To investigate the effect of Survivin on neuronal apoptosis in the substantia nigra in a rat model of Parkinson’s disease（PD）. Methods：Healthy adult male Sprague Dawley rats were randomly divided into 4 groups：control（n=8）；PD（n=8）；PD + adenovirus empty vector（n=8）；and PD + Survivin（n=8）. Following 1 week of model induction，rats from the control and PD group received normal stimulation with no intervention，rats from the PD + adenovirus empty vector group received an empty adenovirus vec－tor injection in the substantia nigra，and rats from the PD + Survivin treatment group received an injection of Survivin adenovirus in the substantia nigra. Behavioral evaluation of spontaneous rotation was performed by injecting apomorphine（APO） every week follow－ing completion of the treatment. Following behavioral testing，brain tissue was collected. Neuronal apoptosis in the substantia nigra was determined by TUNEL staining. qRT-PCR and western blot were used to determine mRNA and protein levels of Caspase 3 and 9，respectively. Results：Compared to the PD + adenovirus empty vector group，the PD + Survivin treatment group exhibited significantly fewer spontaneous rotations following treatment（F=22.315，P=0.000，P=0.000）. Compared with the PD group and the PD + adenovirus empty vector group，neuronal apoptosis was signifi－cantly reduced in the PD + Survivin treatment group（F=38.912，P=0.000，P=0.000）. Compared with the control group，mRNA expression of Caspase-3 and 9（F=338.429，P=0.000，P=0.000；F=283.352，P=0.000，P=0.000） and protein expression of cleaved Caspase-3 and 9（F=640.262，P=0.000，P=0.000；F=354.210，P=0.000，P=0.000） were significantly decreased. Conclusion：Sur－vivin can inhibit apoptosis in the brain by inhibiting Caspase-3 and -9 at the mRNA and protein expression level in rat model of Parkinson’s disease，resulting in protection against neurodegeneration in the substantia nigra and reduced Parkinson’s disease-related symptoms.