Department of Biological Sciences, California State University, Los Angeles, 5151 State University Dr., Los Angeles, CA 90032, USA.
Eur J Pharmacol. 2010 May 10;633(1-3):1-9. doi: 10.1016/j.ejphar.2010.01.012. Epub 2010 Feb 8.
Nitric oxide (NO), physical exercise and/or antidepressant drugs, through the increased release of norepinephrine and brain-derived neurotrophic factor (BDNF), have been shown to exert profound protective, pro-survival effects on neurons otherwise compromised by injury, disease, prolonged stress, and subsequent depression in vivo. We sought, therefore, to evaluate such survival and neuroprotection in hippocampal neurons in culture, which, in an analogous model of in vivo cellular stress, was deprived of several vital nutrients. We assessed pro-survival outcomes following the application of norepinephrine or the noradrenergic partial agonist, clonidine, a general nitric oxide synthase inhibitor and NO donor, using a cell survival assay and quantitative Western blotting of the survival signaling molecules, BDNF, P-CREB, P-Akt, and P-MAPK in hippocampal neuronal lysates. We demonstrate that norepinephrine, clonidine, the NO donor and various combinations of these drugs increased cell survival and the immunoreactivity of the four survival signaling molecules in the face of nutrient deprivation stress, whereas the NO synthase inhibitor, and each of several survival signaling pathway inhibitors all decreased cell survival even below that of controls without nutrient supplementation. These results demonstrate that conditions that make cells vulnerable to environmental/toxic insult can be offset by norepinephrine and its related drugs or by NO donors and exacerbated by drugs that specifically inhibit a key survival signaling pathway. These results indicate that pharmacological intervention can promote neuroprotection and survival signaling in the face of nutrient withdrawal, but that this may require that several pathways remain intact.
一氧化氮（NO）、体育锻炼和/或抗抑郁药物通过增加去甲肾上腺素和脑源性神经营养因子（BDNF）的释放，已被证明对神经元具有深远的保护和生存作用，否则神经元会因损伤、疾病、长期压力和随后的体内抑郁而受损。因此，我们试图在培养的海马神经元中评估这种生存和神经保护作用，在类似于体内细胞应激的模型中，这些神经元被剥夺了几种重要的营养物质。我们通过细胞存活测定和海马神经元裂解物中存活信号分子 BDNF、P-CREB、P-Akt 和 P-MAPK 的定量 Western 印迹，评估了去甲肾上腺素或去甲肾上腺素部分激动剂可乐定、通用一氧化氮合酶抑制剂和 NO 供体应用后的存活结果。我们证明，去甲肾上腺素、可乐定、NO 供体以及这些药物的各种组合在营养剥夺应激下增加了细胞存活和四种存活信号分子的免疫反应性，而 NO 合酶抑制剂以及几种存活信号通路抑制剂中的每一种都降低了细胞存活，甚至低于没有营养补充的对照细胞。这些结果表明，使细胞易受环境/毒性损伤的条件可以被去甲肾上腺素及其相关药物或 NO 供体抵消，而被专门抑制关键存活信号通路的药物加剧。这些结果表明，药理学干预可以在营养物质耗尽的情况下促进神经保护和存活信号，但这可能需要几个途径保持完整。
