Lynch J J, Yu S P, Canzoniero L M, Sensi S L, Choi D W
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri, USA.
J Pharmacol Exp Ther. 1995 Apr;273(1):554-60.
Blockers of voltage-gated Na+ channels can protect central neuronal axons from hypoxic injury in vitro but have shown limited neuroprotective effects on neurons, where substantial injury is mediated by glutamate receptors. We explored the ability of several voltage-gated Na+ channel blockers to protect murine cultured cortical neurons from injury induced by oxygen-glucose deprivation. Whole-cell recordings from neurons revealed two types of Na+ currents activated by membrane depolarization: one rapidly inactivating and the other noninactivating. Both currents were blocked by tetrodotoxin (TTX) and 5,5-diphenylhydantoin (phenytoin). Fluorescent imaging using the Na(+)-selective dye SBFI confirmed that TTX attenuated the increase in intracellular free Na+ induced by oxygen-glucose deprivation. Addition of TTX (1 microM) but not phenytoin (10-100 microM) produced a small and variable reduction in neuronal death subsequent to oxygen-glucose deprivation for 40 to 50 min. Blockade of glutamate neurotoxicity by combined addition of MK-801, 7-chlorokynurenate and 6-cyano-7-nitroquinoxaline-2,3-dione markedly reduced injury such that prolonged deprivation times (75-100 min) were needed to induce widespread neuronal death. In this setting of glutamate receptor blockade, addition of TTX, phenytoin or one of several other Na+ channel blockers--lidocaine (100 microM), QX-314 (1 mM), quinidine (100 microM) or lorcainide (10 or 100 microM)--all further reduced neuronal death. Present results raise the possibility that Na+ channel blockers may be useful in protecting gray matter from hypoxic-ischemic injury, especially when combined with antiexcitotoxic approaches.
电压门控性Na⁺通道阻滞剂在体外可保护中枢神经元轴突免受缺氧损伤,但对神经元的神经保护作用有限,因为神经元的实质性损伤是由谷氨酸受体介导的。我们探究了几种电压门控性Na⁺通道阻滞剂保护小鼠培养皮质神经元免受氧糖剥夺诱导损伤的能力。对神经元进行全细胞记录发现,膜去极化可激活两种类型的Na⁺电流:一种快速失活,另一种非失活。这两种电流均被河豚毒素(TTX)和5,5-二苯基乙内酰脲(苯妥英)阻断。使用Na⁺选择性染料SBFI进行荧光成像证实,TTX可减弱氧糖剥夺诱导的细胞内游离Na⁺的增加。添加TTX(1 μM)而非苯妥英(10 - 100 μM)可使氧糖剥夺40至50分钟后神经元死亡出现小幅度且变化不定的减少。联合添加MK - 801、7 - 氯犬尿氨酸和6 - 氰基 - 7 - 硝基喹喔啉 - 2,3 - 二酮阻断谷氨酸神经毒性可显著减少损伤,以至于需要延长剥夺时间(75 - 100分钟)才能诱导广泛的神经元死亡。在这种谷氨酸受体阻断的情况下,添加TTX、苯妥英或其他几种Na⁺通道阻滞剂之一——利多卡因(100 μM)、QX - 314(1 mM)、奎尼丁(100 μM)或劳卡尼(10或100 μM)——均能进一步减少神经元死亡。目前的结果提示,Na⁺通道阻滞剂可能有助于保护灰质免受缺氧缺血性损伤,尤其是与抗兴奋毒性方法联合使用时。