NADPH oxidases (NOX) are activated in ischemic conditions leading to increases in reactive oxygen species (ROS) and neurotoxicity. The aim of the present study was to investigate the role of NOX in the development of retinal pathologies, associated with excitotoxicity and the evaluation of NOX inhibitors as putative therapeutic agents. Sprague-Dawley rats were used for the induction of the in vivo retinal model of (RS)-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid hydrobromide (AMPA) excitotoxicity. Rats were intravitreally administered with PBS, AMPA (42 nmoles) or AMPA + NOX inhibitors, VAS2870 (pan-NOX inhibitor, 10⁻⁶-10⁻⁴ M), ML171 (NOX1 inhibitor, 10⁻⁵, 10⁻⁴ M), and GLX7013114 (NOX4 inhibitor, 10⁻⁴ M). Immunohistochemical studies were performed using antibodies raised against nitrotyrosine, a ROS/oxidative stress marker, bNOS, a neuronal marker for nitric oxide synthase and the macro and microglia markers, glial fibrillary acidic protein and ionized calcium-binding adaptor molecule-1, respectively. VAS2870 and ML171 showed neuroprotective and anti-inflammatory actions reversing the AMPA induced reduction of bNOS expressing amacrine cells and attenuating macro/microglial activation. GLX7013114 (10⁻⁴ M) did not protect bNOS expressing amacrine cells, but it did attenuate the AMPA induced increase in nitrotyrosine positive cells and activation of glial cells. These results suggest that NOX1, NOX4 and possibly NOX2 (due to the actions of VAS2870) play an important role in the pathophysiology of the retina and that NOX inhibitors are putative neuroprotective and anti-inflammatory agents against retinal abnormalities caused by excitotoxicity.

NADPH氧化酶（NOX）在缺血条件下被激活，导致活性氧（ROS）和神经毒性增加。本研究的目的是调查NOX在视网膜病理学发展中的作用，与兴奋性毒性相关，并评估NOX抑制剂作为推定治疗剂的作用。Sprague-Dawley大鼠用于诱导（RS）-α-氨基-3-羟基-3-羟基-5-甲基-4-异恶唑丙酸氢溴酸（AMPA）兴奋性体内视网膜模型。大鼠玻璃体内用PBS，AMPA（42纳摩尔）或AMPA + NOX抑制剂，VAS2870施用（泛NOX酶抑制剂，10 ^-6 -10 ^-4  M），ML171（NOX1抑制剂，10 ^-5，10 ^-4  M），和GLX7013114（NOX4抑制剂，10^-4  M）。免疫组织化学研究分别使用针对硝基酪氨酸的抗体，ROS /氧化应激标记，bNOS，一氧化氮合酶的神经元标记以及大胶质细胞和小胶质细胞标记，神经胶质纤维酸性蛋白和离子化钙结合衔接子分子-1进行。VAS2870和ML171具有神经保护作用和抗炎作用，可以逆转AMPA诱导的表达bNOS的无长突细胞减少，并减弱巨胶质/小胶质细胞的活化。GLX7013114（10 ^-4 M）不保护表达bNOS的无长突细胞，但是确实减弱了AMPA诱导的硝基酪氨酸阳性细胞增加和神经胶质细胞的活化。这些结果表明，NOX1，NOX4以及可能的NOX2（由于VAS2870的作用）在视网膜的病理生理中起着重要作用，并且NOX抑制剂是针对由兴奋性毒性引起的视网膜异常的假定的神经保护和抗炎剂。