Expression profiling of peripheral blood mononuclear cells in normal controls and patients with Parkinson's disease

Autophagy is a highly conserved degradation pathway whereby not only cytosolic components but also aberrant proteins are sequestered within double-membraned vesicles. Parkinson's disease (PD) is pathologically characterized by accumulation of phosphorylated α-synuclein in the neuronal cytoplasm and its accumulation occurs in the peripheral autonomic nervous system as well as the central nervous system. In the brains of patients with PD, abnormal autophagy is known to occur and be involved with neurodegeneration. To investigate abnormal autophagy in peripheral blood mononuclear cells (PBMC) in the patients with PD, we performed whole transcriptome analysis of PBMC obtained from 9 normal controls and 10 patients with PD. PBMC in 9 normal controls and 10 patients with PD were isolated by Ficoll-paque plus (GE Healthcare, 17144002) centrifugation of heparinized blood and frozen at -80 degrees Celsius until microarray or qRT-PCR.

自噬是一种高度保守的降解途径，可将胞质组分与异常蛋白一并包裹于双层膜囊泡中进行降解。帕金森病（Parkinson's disease, PD）的病理特征为神经元胞质内磷酸化α-突触核蛋白的聚集，且该聚集可同时发生于外周自主神经系统与中枢神经系统。已有研究证实，帕金森病患者脑内存在自噬异常，且该异常与神经退行性病变的发生发展密切相关。为探究帕金森病患者外周血单个核细胞（peripheral blood mononuclear cells, PBMC）中的自噬异常情况，本研究对9名正常对照者与10名帕金森病患者的PBMC进行了全转录组分析。本研究通过Ficoll-paque Plus（通用电气医疗集团GE Healthcare，货号17144002）密度梯度离心法分离肝素化全血中的PBMC，将分离得到的细胞冻存于-80℃环境中，直至开展基因芯片（microarray）或实时定量反转录聚合酶链反应（quantitative real-time reverse transcription polymerase chain reaction, qRT-PCR）检测。