Neuroprotective Effects of Propofol in a Model of Ischemic Cortical Cell Cultures: Role of Glutamate and Its Transporters
Neuroprotective Effects of Propofol in a Model of Ischemic Cortical Cell Cultures: Role of Glutamate and Its Transporters
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DOI：
10.1097/00000542-200308000-00018
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发表时间：
2003-08
期刊：
影响因子：
8.8
通讯作者：
L. Velly;B. Guillet;F. Masmejean;A. Nieoullon;N. Bruder;F. Gouin;P. Pisano
中科院分区：
文献类型：
--
作者：
L. Velly;B. Guillet;F. Masmejean;A. Nieoullon;N. Bruder;F. Gouin;P. Pisano
背景在脑缺血期间，谷氨酸释放过量及其高亲和力转运功能障碍导致细胞外谷氨酸积累，这在神经元死亡中发挥重要作用。作者在缺血性皮层细胞培养模型中研究了异丙酚神经保护作用、谷氨酸细胞外浓度和谷氨酸转运蛋白活性之间的关系。方法 将 13 天大的原代皮层神经元神经胶质细胞培养物置于厌氧室中 90 分钟的缺氧-葡萄糖联合剥夺 (OGD)，然后进行复氧。仅在OGD期间添加异丙酚，其效果与N-甲基-d-天冬氨酸受体拮抗剂地佐西平（MK-801）的效果进行比较。损伤后 24 小时，通过乳酸脱氢酶释放来量化细胞死亡，并通过 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四唑溴化物 (MTT) 的还原来量化细胞活力。在 OGD 期结束时，分别通过高效液相色谱法和 L-[3H]谷氨酸掺入细胞中测定培养物上清液中谷氨酸的细胞外浓度和谷氨酸摄取。结果 在临床相关浓度 (0.05–10 m) 下，异丙酚提供的保护与 MK-801 相当。它显着减少了乳酸脱氢酶的释放并增加了MTT的减少。在缺血性损伤结束时，异丙酚能够逆转 OGD 诱导的谷氨酸细胞外浓度增加和谷氨酸摄取减少。 3-甲基谷氨酸对神经胶质GLT1转运蛋白的抑制并没有进一步改变丙泊酚对谷氨酸摄取的影响，表明GLT1不是丙泊酚的主要靶标。结论 异丙酚在 OGD 体外模型中显示出神经保护作用，这显然是通过不依赖 GLT1 恢复损伤期间受损的谷氨酸摄取来介导的。
Background During cerebral ischemia, excess of glutamate release and dysfunction of its high affinity transport induce an accumulation of extracellular glutamate, which plays an important role in neuronal death. The authors studied the relationship among propofol neuroprotection, glutamate extracellular concentrations, and glutamate transporter activity in a model of ischemic cortical cell cultures. Methods Thirteen-day-old primary cortical neuronal-glial cultures were exposed to a 90-min combined oxygen–glucose deprivation (OGD) in an anaerobic chamber, followed by reoxygenation. Propofol was added only during the OGD period, and its effect was compared to that of the N-methyl-d-aspartate receptor antagonist dizocilpine (MK-801). Twenty-four hours after the injury, cell death was quantified by lactate dehydrogenase release and cell viability by reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). Extracellular concentrations of glutamate in culture supernatants and glutamate uptake were performed at the end of OGD period by high-performance liquid chromatography and incorporation of l-[3H]glutamate into cells, respectively. Results At clinically relevant concentrations (0.05–10 m), propofol offered protection equivalent to that of MK-801. It significantly reduced lactate dehydrogenase release and increased the reduction of MTT. At the end of the ischemic injury, propofol was able to reverse the OGD-induced increase in glutamate extracellular concentrations and decrease of glutamate uptake. The inhibition of the glial GLT1 transporter by 3-methyl-glutamate did not further modify the effect of propofol on glutamate uptake, suggesting that GLT1 was not the major target of propofol. Conclusion Propofol showed a neuroprotective effect in this in vitro model of OGD, which was apparently mediated by a GLT1-independent restoration of the glutamate uptake impaired during the injury.