Transient glucose and amino acid deprivation induces delayed preconditioning in cultured rat cortical neurons
Transient glucose and amino acid deprivation induces delayed preconditioning in cultured rat cortical neurons
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DOI：
10.1111/j.1471-4159.2006.03899.x
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发表时间：
2006-07-01
影响因子：
4.7
通讯作者：
Busija, DW
中科院分区：
文献类型：
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作者：
Gáspár, T;Kis, B;Busija, DW
一些研究表明，缺糖与缺氧或氧化磷酸化抑制相结合，会导致神经元对缺血耐受的发展。我们实验的目的是研究在不缺氧或不抑制电子转移链的情况下，通过瞬时能量剥夺是否可以达到类似的效果。将原代培养的大鼠皮质神经细胞在无葡萄糖、无氨基酸的平衡盐液中加B27在常氧条件下孵育3、6或9h进行预适应。培养24 h后，给予缺氧缺糖、谷氨酸或过氧化氢处理。致死性损伤后24小时测定细胞存活率。还对影响自由基产生的潜在机制进行了研究。能量剥夺可保护神经细胞免受致死性刺激(如未处理组细胞存活率为33.1+/-0.52%，能量剥夺9h组为80.1+/-1.27%)，降低线粒体膜电位，减少自由基形成，减弱谷氨酸受体刺激引起的细胞内游离钙激增，并导致GSH水平升高。我们的发现表明，短暂的能量剥夺导致延迟的预适应，并防止氧化损伤和神经细胞死亡。
Several studies have demonstrated that glucose deprivation, combined either with anoxia or with the inhibition of oxidative phosphorylation, leads to the development of ischemic tolerance in neurons. The aim of our experiments was to investigate whether similar effects could be achieved by transient energy deprivation without either anoxia or the inhibition of the electron transfer chain. Preconditioning was carried out by incubating primary rat cortical neuronal cultures for 3, 6 or 9 h in a glucose- and amino acid-free balanced salt solution supplemented with B27 in normoxic conditions. After 24 h, neuronal cultures were exposed to oxygen-glucose deprivation, glutamate or hydrogen peroxide. Cell viability was measured 24 h after the lethal insults. Potential mechanisms that can influence free radical production were also examined. Energy deprivation protected neuronal cells against lethal stimuli (e.g. cell survival after oxygen-glucose deprivation was 33.1 +/- 0.52% in the untreated group and 80.1 +/- 1.27% in the 9-h energy deprivation group), reduced mitochondrial membrane potential, decreased free radical formation, attenuated the intracellular free calcium surge upon glutamate receptor stimulation, and resulted in an elevated level of GSH. Our findings show that transient energy deprivation induces delayed preconditioning and prevents oxidative injuries and neuronal cell death.