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

描述(申请人提供)：中医药可能为帕金森氏症的治疗提供一些值得西医进一步关注的新方法。一个最好的例子是电针，这是一种众所周知的中医，已被用作帕金森氏症患者的替代疗法，并取得了一些成功。我们建议在凯斯西储大学、匹兹堡大学和中国医科大学三个机构之间建立一个合作项目，以启动一个关于针灸作为帕金森病干预措施的研究项目。一般假设：电针通过激活神经营养因子(NTF)的基因表达，从而降低DA神经元对环境和遗传因素的易感性，从而降低这些神经元对内质网应激和线粒体功能障碍的易感性。这款R21有三个具体目标。目的1：在美国实验室建立MPTP处理小鼠的电针神经保护模型。在第一个实验中，我们将使用C57/b16小鼠和进行性MPTP模型。实验分为六组：(1)急性MPTP+100 Hz电针，(2)MPTP+0 Hz针刺，(3)MPTP+电针无效部位，(4)MPTP单独治疗，(5)赋形剂治疗组小鼠接受电针治疗，(6)赋形剂治疗组小鼠加假针刺。在2.5周和5.5周后，将使用行为、组织学和生化分析来评估神经保护。目的：探讨电针对内质网应激指标和线粒体功能的影响。利用实验1和实验2中产生的组织，我们将使用生化和免疫组织化学方法评估内质网应激下游靶标的激活、UPR靶基因的蛋白上调以及线粒体参数。目的3：确定GDNF的可获得性与电针反应之间是否存在交互作用。两组不同的GDNF缺失突变体将被MPTP急性处理，接受电针或假处理，然后评估电针诱导的神经保护。

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.