Foster Steven B, Wrona Monika Z, Han Jilin, Dryhurst Glenn
Department of Chemistry and Biochemistry, University of Oklahoma, Norman, Oklahoma 73019-0370, USA.
Chem Res Toxicol. 2003 Oct;16(10):1372-84. doi: 10.1021/tx030015l.
Reactive oxygen species (ROS) and reactive nitrogen species (RNS), particularly peroxynitrite, have been implicated as key participants in the dopaminergic neurotoxicity of 1-methyl-4-phenylpyridinium (MPP(+)). However, on the basis of available information, it is not clear whether the MPP(+)-induced overproduction of ROS and RNS occurs in the intraneuronal and/or extracellular compartment. Early steps in the neurotoxic mechanism evoked by MPP(+) include a profound dopaminergic energy impairment, which mediates a massive release of dopamine (DA), glutathione (GSH), and cysteine (CySH). In the event that MPP(+) mediates extracellular generation of ROS (such as superoxide and/or hydroxyl radicals) and/or peroxynitrite, released DA, GSH, and CySH should be oxidized forming thioethers of DA and disulfides. Using microdialysis experiments in which MPP(+) was perfused into the striatum of awake rats, the present study was unable to detect the presence of such biomarkers of extracellular ROS and/or RNS generation. However, MPP(+) induced a transient, concentration-dependent rise of extracellular l-3,4-dihydroxyphenylalanine (l-DOPA), identified on the basis of dialysate analysis using several HPLC methods and its conversion to DA by purified l-DOPA decarboxylase (DDC). Methamphetamine (30 mg/kg, i.p.) similarly caused a significant but transient rise of l-DOPA in the rat striatum. Antioxidants such as salicylate and mannitol had no effect on the MPP(+)-mediated elevation of extracellular l-DOPA, suggesting that it is not formed by nonenzymatic hydroxylation of l-tyrosine by ROS or RNS. Rather, in vivo, but not in vitro, MPP(+) caused rapid inhibition of DDC, which appears to result in intraneuronal accumulation and subsequent release of l-DOPA. Because l-DOPA can mediate l-glutamate release, as well as be an excitotoxin, the possibility is raised that l-DOPA may play a role in the dopaminergic neurotoxicity of MPP(+).
活性氧(ROS)和活性氮(RNS),尤其是过氧亚硝酸盐,被认为是1-甲基-4-苯基吡啶离子(MPP(+))多巴胺能神经毒性的关键参与者。然而,根据现有信息,尚不清楚MPP(+)诱导的ROS和RNS过量产生是发生在神经元内和/或细胞外区室。MPP(+)诱发的神经毒性机制的早期步骤包括严重的多巴胺能能量损伤,这介导了多巴胺(DA)、谷胱甘肽(GSH)和半胱氨酸(CySH)的大量释放。如果MPP(+)介导细胞外ROS(如超氧化物和/或羟基自由基)和/或过氧亚硝酸盐的产生,释放的DA、GSH和CySH应该被氧化形成DA的硫醚和二硫化物。通过将MPP(+)灌注到清醒大鼠纹状体的微透析实验,本研究未能检测到细胞外ROS和/或RNS产生的此类生物标志物的存在。然而,MPP(+)诱导细胞外l-3,4-二羟基苯丙氨酸(l-DOPA)出现短暂的、浓度依赖性升高,这是通过使用几种高效液相色谱方法对透析液进行分析并通过纯化的l-DOPA脱羧酶(DDC)将其转化为DA来确定的。甲基苯丙胺(30mg/kg,腹腔注射)同样引起大鼠纹状体中l-DOPA显著但短暂的升高。抗氧化剂如水杨酸盐和甘露醇对MPP(+)介导的细胞外l-DOPA升高没有影响,这表明它不是由ROS或RNS对l-酪氨酸的非酶促羟基化形成的。相反,在体内而非体外,MPP(+)导致DDC快速抑制,这似乎导致l-DOPA在神经元内积累并随后释放。由于l-DOPA可以介导l-谷氨酸释放,并且还是一种兴奋性毒素,因此l-DOPA可能在MPP(+)的多巴胺能神经毒性中起作用。