Pearce R K, Owen A, Daniel S, Jenner P, Marsden C D
Neurodegenerative Diseases Research Centre, King's College, London, United Kingdom.
J Neural Transm (Vienna). 1997;104(6-7):661-77. doi: 10.1007/BF01291884.
Depletion of reduced glutathione occurs in the substantia nigra in Parkinson's disease and in incidental Lewy body disease (presymptomatic Parkinson's disease) which may implicate oxidative stress in the neurodegenerative process. In this study mercury orange fluorescent staining and immunostaining with an antibody to reduced glutathione have been used to determine the distribution of reduced glutathione in the substantia nigra in Parkinson's disease compared with normal individuals. Mercury orange staining showed moderate background levels of fluorescence in the neuropil in both control and Parkinson's disease substantia nigra and localised reduced glutathione to the somata of melanized nigral neurons and glial elements of the neuropil. Neuronal nuclei revealed a relative lack of fluorescence after mercury orange staining. There was a significant depletion of reduced glutathione in surviving neurons in Parkinson's disease compared to nerve cell populations in control tissue. Mercury orange fluorescence indicated a high concentration of reduced glutathione in a subpopulation of non-neuronal cells, most likely astrocytes or microglia. Immunohistochemical examination of nigral tissue from the same Parkinson's disease and control patients with an antibody to glutathione showed staining in neuronal perikarya and axonodendritic processes of melanized nigral neurons which was generally most intense in control neurons. Moderately intense staining of the background neuropil, most prominent in control nigras, and staining of capillary walls was also detected. Intense staining was seen in cells with the morphological features of glial cells in both control and PD nigra. These data show a significant presence of reduced glutathione in the cell bodies and axons of nigral neurons. They are in agreement with biochemical studies showing depletion of reduced glutathione in substantia nigra in Parkinson's disease, and indicate a significant loss of neuronal reduced glutathione in surviving nigral neurons in Parkinson's disease.
帕金森病及偶发性路易体病（症状前帕金森病）患者黑质中还原型谷胱甘肽耗竭，这可能与神经退行性变过程中的氧化应激有关。在本研究中，与正常个体相比，采用汞橙荧光染色和抗还原型谷胱甘肽抗体免疫染色来确定帕金森病黑质中还原型谷胱甘肽的分布。汞橙染色显示，在对照和帕金森病黑质的神经毡中荧光背景水平适中，且还原型谷胱甘肽定位于黑素化黑质神经元的胞体及神经毡的胶质成分。汞橙染色后神经元细胞核显示相对缺乏荧光。与对照组织中的神经细胞群体相比，帕金森病存活神经元中的还原型谷胱甘肽显著耗竭。汞橙荧光表明非神经元细胞亚群中还原型谷胱甘肽浓度较高，最可能是星形胶质细胞或小胶质细胞。用谷胱甘肽抗体对同一帕金森病患者和对照患者的黑质组织进行免疫组化检查，结果显示黑素化黑质神经元的神经元胞体和轴突树突过程中有染色，在对照神经元中通常最为强烈。还检测到背景神经毡有中度强烈染色，在对照黑质中最为突出，以及毛细血管壁有染色。在对照和帕金森病黑质中，具有胶质细胞形态特征的细胞均可见强烈染色。这些数据表明黑质神经元的细胞体和轴突中存在大量还原型谷胱甘肽。它们与生化研究结果一致，生化研究表明帕金森病黑质中还原型谷胱甘肽耗竭，并表明帕金森病存活黑质神经元中神经元还原型谷胱甘肽显著丧失。
