Rosenfeld J, Cook S, James R
Department of Neurology, Emory University, Atlanta, Georgia 30322, USA.
Exp Neurol. 1997 Sep;147(1):37-47. doi: 10.1006/exnr.1997.6604.
Oxidative injury has been implicated in the pathophysiology of neuronal injury and neurodegenerative disease. Antioxidant proteins provide an endogenous defense against such oxidative injury and may yield important clues to mechanisms of cytoprotection and neuronal recovery. Axotomy is the simplest model of neuronal injury and lesioning the sciatic nerve allows concurrent study of both motor (spinal cord) and sensory (dorsal root ganglia, DRG) neurons affected by the same injury. This study was designed to examine the expression of superoxide dismutase (SOD), an essential antioxidant protein, in motor and sensory neurons following complete axotomy of peripheral nerve. Immunocytochemical, quantitative immunoblot, and enzymatic activity assay techniques are used. By 12 days after axotomy, immunocytochemical expression of Mn-SOD is markedly increased in affected DRG and spinal cord. A similar increase in Cu/Zn-SOD is not seen in DRG or spinal cord. This immunocytochemical staining is associated with a significant increase in specific activity and Mn-SOD protein content as measured on quantitative immunoblots. This report suggests, for the first time, that Mn-SOD and not Cu/Zn-SOD increases in sensory neurons of the DRG and motor neurons of the spinal cord following distal axotomy of the sciatic nerve. Quantitative measurements of Mn-SOD following axotomy reveals that the increase in immunocytochemical reactivity is associated with an approximately 30% increase in specific activity when comparing lesioned and contralateral spinal cord samples. These data suggest that Mn-SOD may have a more significant role in the pathophysiology of neuronal injury than Cu/Zn-SOD.
氧化损伤与神经元损伤和神经退行性疾病的病理生理学有关。抗氧化蛋白为抵御此类氧化损伤提供了内源性防御,并且可能为细胞保护和神经元恢复机制提供重要线索。轴突切断术是神经元损伤的最简单模型,切断坐骨神经可同时研究受同一损伤影响的运动(脊髓)和感觉(背根神经节,DRG)神经元。本研究旨在检查外周神经完全切断后,运动和感觉神经元中一种重要的抗氧化蛋白超氧化物歧化酶(SOD)的表达。采用免疫细胞化学、定量免疫印迹和酶活性测定技术。轴突切断术后12天,受影响的DRG和脊髓中Mn-SOD的免疫细胞化学表达明显增加。在DRG或脊髓中未观察到Cu/Zn-SOD有类似增加。这种免疫细胞化学染色与定量免疫印迹上测得的比活性和Mn-SOD蛋白含量的显著增加相关。本报告首次表明,坐骨神经远端切断术后,DRG的感觉神经元和脊髓的运动神经元中Mn-SOD增加,而不是Cu/Zn-SOD增加。轴突切断术后对Mn-SOD的定量测量显示,与对侧脊髓样本相比,免疫细胞化学反应性的增加与比活性增加约30%相关。这些数据表明,在神经元损伤的病理生理学中,Mn-SOD可能比Cu/Zn-SOD发挥更重要的作用。