Department of Dental Hygiene, Division of Health Science, Dongseo University, Busan, Korea.
J Neurosci Res. 2011 Jan;89(1):96-107. doi: 10.1002/jnr.22525.
During seizures, glucose concentrations are high in the hippocampus. Mitochondrial hexokinase (HK) catalyzes the first essential step of glucose metabolism and directly couples extramitochondrial glycolysis to intramitochondrial oxidative phosphorylation. The neuroprotective effects of an HK inhibitor, 3-bromopyruvate (3-BrPA), on kainic acid (KA)-induced excitotoxic injury were investigated. Hippocampal slices were prepared from hippocampi of 6-8-day-old rats using a tissue chopper and placed on a membrane insert. After a treatment with KA (5 μM) for 15 hr, neuronal death was quantified by propidium iodide (PI), cresol violet, and TUNEL staining. KA-induced cell death was significantly prevented by 30 μM 3-BrPA treatment. According to Western blots, the expression level of phospho-Akt increased after 3-BrPA treatment. The induction of long-term potentiation (LTP) at 48 hr after 3-BrPA treatment tended to increase in the CA1 area compared with the KA-only group, but the difference was not significant. Blocking the PI3 kinase/Akt pathway using LY294002 reversed the neuroprotective effect of 3-BrPA. These results suggest that inhibition of HK may play a protective role against neuronal death in KA-induced excitotoxic injury.
在癫痫发作期间，海马体中的葡萄糖浓度很高。线粒体己糖激酶（HK）催化葡萄糖代谢的第一步，直接将细胞外糖酵解与线粒体氧化磷酸化偶联。本研究旨在探讨 HK 抑制剂 3-溴丙酮酸（3-BrPA）对红藻氨酸（KA）诱导的兴奋性毒性损伤的神经保护作用。采用组织切片机从 6-8 日龄大鼠的海马体中制备海马切片，并置于膜插入物上。用 5 μM KA 处理 15 小时后，通过碘化丙啶（PI）、甲酚紫和 TUNEL 染色定量神经元死亡。30 μM 3-BrPA 处理可显著预防 KA 诱导的细胞死亡。根据 Western blot，3-BrPA 处理后磷酸化 Akt 的表达水平增加。与仅用 KA 处理的组相比，3-BrPA 处理后 48 小时 CA1 区的长时程增强（LTP）诱导趋势增加，但差异无统计学意义。使用 LY294002 阻断 PI3 激酶/Akt 通路可逆转 3-BrPA 的神经保护作用。这些结果表明，抑制 HK 可能在 KA 诱导的兴奋性毒性损伤中发挥保护神经元的作用。
