丘脑底核电刺激对帕金森大鼠黑质神经元线粒体的保护作用及机制研究

Selective loss of dopaminergic neurons in the subtantian nigra pars compacta (SNc) is the important etiological basis of Parkinson’s diseases（PD）. Mitochondrial dysfunction plays important roles in the development of loss of dopaminergic neurons in SNc. Neuroprotective effect of deep brain stimulation of the subthalamic nucleus (STN-DBS) has been demonstrated in researches of PD, yet the mechanism is still unclear. Our previous study results demonstrated that STN-DBS could improve the 6-hydroxydopamine induced mitochondrial lesion. Thus, we propose that STN-DBS protects substantia nigra dopaminergic neuron by inhibiting mitochondrial dysfunction. In this study, we will establish a rat model of PD and implant electrical stimulator in the PD rat model. We will use methods of Nissl staining, Tunel staining, immunohistochemical staining, transmission electron microscope, immofluorescence and western-blot to observe the motochondrial morphology, motochondrial function and mitochondrial related apoptotic protein in SNc. The results will explore the neuroprotective mechanism of STN-DBS on mitochondria of SNc.

中脑黑质多巴胺能神经元进行性凋亡是帕金森病(Parkinson’s Disease, PD)发生发展的根本原因，而线粒体功能障碍在多巴胺能神经元凋亡过程中起关键作用。丘脑底核脑深部电刺激（subthalamic nucleus brain stimulation, STN-DBS）神经保护作用在治疗PD的研究中已经证实，但其确切保护机制仍然不清。前期研究我们发现STN-DBS可以抑制6-羟基多巴胺（6-OHDA）引起的PD大鼠中脑黑质线粒体的损伤。因而，我们提出STN-DBS可能是通过稳定线粒体的功能来保护黑质多巴胺神经元。本研究拟建立PD大鼠模型并植入大鼠电刺激器，利用尼氏染色、Tunel染色、免疫组化、透射电子显微镜、免疫荧光及免疫印迹等技术手段，观察STN-DBS对黑质线粒体形态、功能及线粒体相关凋亡蛋白影响，探讨STN-DBS对6-OHDA引起的黑质线粒体损伤的保护作用及机制。

项目背景：探讨丘脑底核对6-羟基多巴胺（6-OHDA）致帕金森大鼠黑质神经元的保护作用及机制。.主要研究内容： SD大鼠随机分为4组，正常对照组、模型组、假刺激组及模型组。模型组、假刺激组及电刺激组于左侧前脑内侧束注射6-OHDA制备大鼠偏侧PD模型，刺激组大鼠左侧丘脑底核区植入刺激电极进行高频电刺激，对四组动物进行行为学、免疫组化、电镜及Western-blot分析。.重要结果：帕金森大鼠在阿扑吗啡诱导后出现明显的旋转行为，且在转棒上的滞留时间缩短，黑质区多巴胺阳性细胞数明显减少，线粒体结构紊乱，线粒体凋亡因子明显增加，而丘脑脑底核电刺激明显改善大鼠的行为学异常，增加黑质多巴胺神经元阳性细胞数；稳定线粒体结构，抑制线粒体凋亡因子的释放。.科学意义：丘脑底核电刺激对6-OHDA致PD大鼠的黑质神经元细胞具有神经保护作用，其机制与丘脑底核电刺激通过保护线粒体来抑制黑质神经元细胞的凋亡有关。