Aguilera Penélope, Chánez-Cárdenas María Elena, Floriano-Sánchez Esaú, Barrera Diana, Santamaría Abel, Sánchez-González Dolores Javier, Pérez-Severiano Francisca, Pedraza-Chaverrí José, Jiménez Perla Deyanira Maldonado
Laboratorio de Patología Vascular Cerebral, Instituto Nacional de Neurología y Neurocirugía Manuel Velasco Suárez, México, DF 14269, Mexico.
Neurotoxicology. 2007 Nov;28(6):1200-7. doi: 10.1016/j.neuro.2007.07.010. Epub 2007 Aug 6.
Excitotoxicity and oxidative stress are mechanisms involved in the neuronal cell death induced by the intrastriatal injection of quinolinic acid (QUIN) as a model of Huntington's disease. Production of nitric oxide by nitric oxide synthase (NOS) has been proposed to participate in QUIN-induced neurotoxicity; however, the precise role of NOS in QUIN-induced toxicity still remains controversial. In order to provide further information on the role of NOS isoforms in QUIN toxicity, we performed real time RT-PCR and immunohistochemistry of inducible NOS (iNOS), endothelial NOS (eNOS) and neuronal NOS (nNOS) and determined Ca(2+)-dependent and Ca(2+)-independent NOS activity in a temporal course (3-48h), after an intrastriatal injection of QUIN to rats. NOS isoforms exhibited a transitory expression of mRNA and protein after QUIN infusion: eNOS increased between 3 and 24h, iNOS between 12 and 24h, while nNOS at 35 and 48h. Ca(2+)-independent activity (iNOS) did not show any change, while Ca(2+)-dependent activity (constitutive NOS: eNOS/nNOS) exhibited increased levels at 3h. Our results support the participation of Ca(2+)-dependent NOS isoforms during the toxic events produced at early times after QUIN injection.
兴奋性毒性和氧化应激是纹状体内注射喹啉酸(QUIN)诱导神经元细胞死亡的机制,QUIN注射是亨廷顿病的一种模型。一氧化氮合酶(NOS)产生的一氧化氮已被认为参与了QUIN诱导的神经毒性;然而,NOS在QUIN诱导的毒性中的确切作用仍存在争议。为了进一步了解NOS同工型在QUIN毒性中的作用,我们对大鼠纹状体内注射QUIN后,在3至48小时的时间进程中进行了诱导型NOS(iNOS)、内皮型NOS(eNOS)和神经元型NOS(nNOS)的实时逆转录聚合酶链反应(RT-PCR)和免疫组织化学检测,并测定了Ca(2+)依赖性和Ca(2+)非依赖性NOS活性。QUIN注入后,NOS同工型表现出mRNA和蛋白质的短暂表达:eNOS在3至24小时增加,iNOS在12至24小时增加,而nNOS在35和48小时增加。Ca(2+)非依赖性活性(iNOS)没有任何变化,而Ca(2+)依赖性活性(组成型NOS:eNOS/nNOS)在3小时时水平升高。我们的结果支持Ca(2+)依赖性NOS同工型在QUIN注射后早期产生的毒性事件中发挥作用。