Zhang J, Perry G, Smith M A, Robertson D, Olson S J, Graham D G, Montine T J
Departments of Pathology, Medicine, and Pharmacology, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Am J Pathol. 1999 May;154(5):1423-9. doi: 10.1016/S0002-9440(10)65396-5.
Oxidative damage, including modification of nucleic acids, may contribute to dopaminergic neurodegeneration in the substantia nigra (SN) of patients with Parkinson's disease (PD). To investigate the extent and distribution of nucleic acid oxidative damage in these vulnerable dopaminergic neurons, we immunohistochemically characterized a common product of nucleic acid oxidation, 8-hydroxyguanosine (8OHG). In PD patients, cytoplasmic 8OHG immunoreactivity was intense in neurons of the SN, and present to a lesser extent in neurons of the nucleus raphe dorsalis and oculomotor nucleus, and occasionally in glia. The proportion of 8OHG immunoreactive SN neurons was significantly greater in PD patients compared to age-matched controls. Midbrain sections from patients with multiple system atrophy-Parkinsonian type (MSA-P) and dementia with Lewy bodies (DLB) also were examined. These showed increased cytoplasmic 8OHG immunoreactivity in SN neurons in both MSA-P and DLB compared to controls; however, the proportion of positive neurons was significantly less than in PD patients. The regional distribution of 8OHG immunoreactive neurons within the SN corresponded to the distribution of neurodegeneration for these three diseases. Nuclear 8OHG immunoreactivity was not observed in any individual. The type of cytoplasmic nucleic acid responsible for 8OHG immunoreactivity was analyzed by preincubating midbrain sections from PD patients with RNase, DNase, or both enzymes. 8OHG immunoreactivity was substantially diminished by either RNase or DNase, and completely ablated by both enzymes. These results suggest that oxidative damage to cytoplasmic nucleic acid is selectively increased in midbrain, especially the SN, of PD patients and much less so in MSA-P and DLB patients. Moreover, oxidative damage to nucleic acid is largely restricted to cytoplasm with both RNA and mitochondrial DNA as targets.
氧化损伤,包括核酸修饰,可能导致帕金森病(PD)患者黑质(SN)中的多巴胺能神经变性。为了研究这些易损多巴胺能神经元中核酸氧化损伤的程度和分布,我们通过免疫组织化学方法对核酸氧化的常见产物8-羟基鸟苷(8OHG)进行了表征。在PD患者中,SN神经元的细胞质8OHG免疫反应强烈,在中缝背核和动眼神经核的神经元中程度较轻,偶尔在神经胶质细胞中也有出现。与年龄匹配的对照组相比,PD患者中8OHG免疫反应性SN神经元的比例显著更高。我们还检查了多系统萎缩帕金森型(MSA-P)和路易体痴呆(DLB)患者的中脑切片。与对照组相比,MSA-P和DLB患者的SN神经元中细胞质8OHG免疫反应性均增加;然而,阳性神经元的比例明显低于PD患者。SN内8OHG免疫反应性神经元的区域分布与这三种疾病的神经变性分布相对应。在任何个体中均未观察到细胞核8OHG免疫反应性。通过用核糖核酸酶(RNase)、脱氧核糖核酸酶(DNase)或两种酶对PD患者的中脑切片进行预孵育,分析了导致8OHG免疫反应性的细胞质核酸类型。单独使用RNase或DNase均可使8OHG免疫反应性显著降低,两种酶同时使用则可使其完全消除。这些结果表明,PD患者中脑尤其是SN中细胞质核酸的氧化损伤选择性增加,而MSA-P和DLB患者中则少得多。此外,核酸的氧化损伤主要局限于细胞质,RNA和线粒体DNA均为靶点。