Hegarty Shane V, Sullivan Aideen M, O'Keeffe Gerard W
Department of Anatomy and Neuroscience, Biosciences Institute, University College Cork, Cork, Ireland.
The Irish Centre for Fetal and Neonatal Translational Research (INFANT), Cork University Maternity Hospital, Cork, Ireland.
J Biol Methods. 2016 Jul 25;3(3):e50. doi: 10.14440/jbm.2016.124. eCollection 2016.
Parkinson's disease (PD) is a neurodegenerative disease that is characterized by motor and non-motor symptoms which result from the progressive degeneration of nigrostriatal ventral midbrain (VM) dopaminergic (DA) neurons, as well as peripheral sympathetic neurons. PD is incurable, with current therapeutic strategies providing symptomatic relief. Neurotrophic factor (NTF) therapy has the potential to protect degenerating neurons in PD. However, there has been limited success in PD clinical trials due to neurotrophic strategies that are invasive, inefficient in delivering sustained neurotrophic support, do not protect all degenerating neurons and may have a compromised mechanism of action in the PD brain. Therefore, while neurotrophic therapy remains a promising disease-modifying approach for PD, it is important to identify novel neurotrophic strategies that can protect all neurons affected by PD. To address this need, we report an integrated approach for pre-clinical evaluation of potential neurotrophic strategies, , pharmacological agents (, drugs/small molecules), signaling proteins (, morphogens) and/or genetic (gene/mRNA) modifications, in cellular models of the neuronal populations that are affected by PD. Herein, we describe, in detail, an protocol that allows a step-wise evaluation of the efficacy, and mechanism(s) of action, of novel neurotrophic strategies in VM DA neurons and sympathetic neurons, following an initial evaluation in a human cell line model of these cells, SH-SY5Y cells. The protocol uses the induction of neurite growth as the primary measure of neurotrophic action. Indeed, the neuro-protection/-restoration of PD-affected axons is widely thought to be an appropriate target for effective therapeutic intervention in PD.
帕金森病（PD）是一种神经退行性疾病，其特征在于运动和非运动症状，这些症状是由黑质纹状体腹侧中脑（VM）多巴胺能（DA）神经元以及外周交感神经元的进行性退化引起的。PD无法治愈，目前的治疗策略只能缓解症状。神经营养因子（NTF）疗法有可能保护PD中退化的神经元。然而，由于神经营养策略具有侵入性、提供持续神经营养支持的效率低下、不能保护所有退化的神经元，并且在PD大脑中的作用机制可能受损，因此PD临床试验的成功率有限。因此，虽然神经营养疗法仍然是一种有前景的PD疾病修饰方法，但识别能够保护所有受PD影响神经元的新型神经营养策略非常重要。为了满足这一需求，我们报告了一种综合方法，用于在受PD影响的神经元群体的细胞模型中对潜在的神经营养策略、药理剂（药物/小分子）、信号蛋白（形态发生素）和/或基因（基因/mRNA）修饰进行临床前评估。在此，我们详细描述了一种方案，该方案允许在这些细胞的人细胞系模型SH-SY5Y细胞中进行初步评估后，逐步评估新型神经营养策略在VM DA神经元和交感神经元中的疗效和作用机制。该方案使用神经突生长的诱导作为神经营养作用的主要衡量标准。事实上，广泛认为对受PD影响的轴突进行神经保护/恢复是PD有效治疗干预的合适靶点。
