Role of GDNF, ER stress and mitochondrial function in effects of acupuncture in models of parkinsonism

药物描述（申请人提供）：中药可能为帕金森病的治疗提供一些新的方法，值得西医进一步关注。一个主要的例子是电针，这是一种众所周知的中医疗法，已被用作PD患者的替代疗法，并取得了一定的成功。我们建议在凯斯西储大学（CWRU），匹兹堡大学（Pitt）和中国医科大学（CMU）三个机构之间建立合作，启动针灸作为PD干预的研究计划。一般假设：EA通过激活包括GDNF在内的神经营养因子（NTFs）的基因表达，降低DA神经元对环境和遗传因素的脆弱性，从而降低这些神经元对ER应激和线粒体功能障碍的易感性。R21有三个具体目标。目的1：在我们的美国实验室中建立一个在MPTP处理的小鼠中的EA神经保护的良好表征模型。在第一个实验中，我们将使用C57/b16小鼠和进行性MPTP模型。将涉及六个小组：（1）急性MPTP加100 Hz EA，（2）MPTP加0 Hz针灸，（3）MPTP加无效部位EA，（4）单独MPTP治疗，（5）给予EA的载体治疗小鼠，和（6）载体治疗小鼠加假针灸。在2.5周和5.5周后，将使用行为、组织学和生物化学分析来评估神经保护。目的2：探讨电针对内质网应激指标和线粒体功能的影响。使用实验1和2中产生的组织，我们将使用生物化学和免疫组织化学方法评估ER应激的下游靶点的激活、UPR靶基因的蛋白上调和线粒体参数。目的3：确定是否有GDNF的可用性之间的相互作用对EA的反应。将两组不同的GDNF无效突变体用MPTP急性处理，进行EA或假处理，然后评估EA诱导的神经保护作用。

DESCRIPTION (provided by applicant): Traditional Chinese Medicine (TCM) may provide some novel approaches to Parkinson's Disease treatment that deserve further attention by Western medicine. A prime example is electroacupuncture, a well-known type of TCM that has been used as an alternative therapy in patients with PD with some success. We propose to establish a collaboration among three institutions, Case Western Reserve University (CWRU), the University of Pittsburgh (Pitt), and China Medical University (CMU), to initiate a research program on acupuncture as an intervention for PD. General hypothesis: EA reduces the vulnerability of DA neurons to both environmental and genetic factors by activating gene expression for neurotrophic factors (NTFs) including GDNF, which then reduces the susceptibility of these neurons to ER stress and mitochondrial dysfunction. This R21 has three specific aims. Aim 1: To establish in our U.S. labs a well-characterized model of EA neuroprotection in MPTP-treated mice. In the first experiment we will use C57/b16 mice and a progressive MPTP model. Six groups will be involved: (1) acute MPTP plus 100 Hz EA, (2) MPTP plus 0 Hz acupuncture, (3) MPTP plus EA at a non-effective site, (4) MPTP treatment alone, (5) vehicle-treated mice given EA, and (6) vehicle treated mice plus sham acupuncture. Behavioral, histological and biochemical analysis will be used to assess neuroprotection after 2.5 and 5.5 weeks. Aim 2: To determine the influence of EA on indices of ER stress and mitochondrial function. Using tissue generated in Experiments 1 and 2, we will assess activation of downstream targets of ER stress, protein upregulation of UPR target genes and mitochondrial parameters using biochemical and immunohistochemical methods. Aim 3: To determine if there is an interaction between GDNF availability on the response to EA. Two different groups of GDNF null mutants will be treated acutely with MPTP, subjected to EA or sham treatments, and then EA-induced neuroprotection assessed.