Vincent Andrea M, Stevens Martin J, Backus Carey, McLean Lisa L, Feldman Eva L
Department of Neurology, University of Michigan, Ann Arbor, MI 48109, USA.
Antioxid Redox Signal. 2005 Nov-Dec;7(11-12):1494-506. doi: 10.1089/ars.2005.7.1494.
The concept that oxidative stress is a key mediator of nerve injury in diabetes has led us to design therapies that target oxidative stress mechanisms. Using an in vitro model of glucose-treated dorsal root ganglion (DRG) neurons in culture, we can examine both free radical generation, using fluorimetric probes for reactive oxygen species, and cell death via the TUNEL assay. The cell culture system is scaled down to a 96-well plate format, and so is well suited to high-throughput screening. In the present study, we test the ability of three drugs, nicotinamide, allopurinol, and alpha-lipoic acid, alone and in combination to prevent DRG neuron oxidative stress and cell death. This combination of drugs is currently in clinical trial in type 1 diabetic patients. We demonstrate independent effects on oxidative stress and neuronal survival for the three drugs, and neuronal protection using the three drugs in combination. The data strengthen the rationale for the current clinical trial. In addition, we describe an effective tool for rapid preclinical testing of novel therapies against diabetic neuropathy.
氧化应激是糖尿病神经损伤关键介质的这一概念促使我们设计针对氧化应激机制的疗法。利用体外培养的葡萄糖处理背根神经节（DRG）神经元模型，我们既能使用荧光探针检测活性氧来研究自由基生成，又能通过TUNEL检测法研究细胞死亡情况。细胞培养系统被缩小至96孔板形式，因此非常适合高通量筛选。在本研究中，我们测试了三种药物烟酰胺、别嘌醇和α -硫辛酸单独及联合使用预防DRG神经元氧化应激和细胞死亡的能力。这种药物组合目前正在1型糖尿病患者中进行临床试验。我们证明了这三种药物对氧化应激和神经元存活具有独立作用，且联合使用这三种药物具有神经元保护作用。这些数据强化了当前临床试验的理论依据。此外，我们描述了一种针对新型糖尿病神经病变疗法进行快速临床前测试的有效工具。
