Department of Anesthesiology, Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Neuroreport. 2021 Mar 3;32(4):312-320. doi: 10.1097/WNR.0000000000001593.
Ischemic stroke is the most frequent cause of long-term morbidity and mortality in the elderly worldwide. Mild hypothermia (32-35°C) has been found to have a neuroprotective effect against ischemic stroke. However, the protective mechanisms remain unclear. In the present study, we explore the neuroprotective effect of mild hypothermia in neuron-astrocyte cocultures by oxygen-glucose deprivation/reoxygenation (OGD/R) as well as the underlying mechanisms. Thionin staining was performed and cell viability, extracellular glutamate concentration and the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway-related proteins were detected after OGD/R. The results indicated that mild hypothermia significantly alleviated damage to Nissl bodies and increased the viability of neurons, which alleviated OGD/R-triggered neuronal injury. Furthermore, mild hypothermia significantly enhanced the phosphorylation of Akt (pAkt) and glutamate transporter-1 (GLT-1) and reduced extracellular glutamate concentration after OGD/R. When the PI3K inhibitor LY294002 was added, neuronal viability and the expression of pAkt and GLT-1 decreased, and extracellular glutamate concentration increased. The protective effect of mild hypothermia was counteracted by LY294002. There was no significant change in neuronal viability or the expression of pAkt and GLT-1 in the group treated with dihydrokainate, an inhibitor of GLT-1-function, compared with the mild hypothermia + OGD/R (HOGD) group, but extracellular glutamate concentration was increased. Consequently, mild hypothermia promoted glutamate clearance by regulating GLT-1 expression via the PI3K/Akt pathway, providing a neuroprotective effect against OGD/R injury.
缺血性脑卒中是全球老年人长期发病率和死亡率的最主要原因。研究发现,轻度体温降低(32-35°C)对缺血性脑卒中具有神经保护作用。然而,其保护机制尚不清楚。本研究通过氧葡萄糖剥夺/复氧(OGD/R)探讨了轻度体温降低对神经元-星形胶质细胞共培养的神经保护作用及其潜在机制。OGD/R 后进行硫堇染色和细胞活力、细胞外谷氨酸浓度以及磷酸肌醇-3-激酶/蛋白激酶 B(PI3K/Akt)通路相关蛋白的检测。结果表明,轻度体温降低可显著减轻 Nissl 小体损伤,增加神经元活力,从而减轻 OGD/R 引起的神经元损伤。此外,轻度体温降低可显著增强 Akt(pAkt)和谷氨酸转运体-1(GLT-1)的磷酸化,降低 OGD/R 后的细胞外谷氨酸浓度。加入 PI3K 抑制剂 LY294002 后,神经元活力以及 pAkt 和 GLT-1 的表达降低,细胞外谷氨酸浓度升高。LY294002 拮抗了轻度体温降低的保护作用。与轻度体温降低+OGD/R(HOGD)组相比,GLT-1 功能抑制剂二氢奎宁酸处理组神经元活力以及 pAkt 和 GLT-1 的表达无明显变化,但细胞外谷氨酸浓度升高。因此,轻度体温降低通过调节 PI3K/Akt 通路促进 GLT-1 表达,从而促进谷氨酸清除,对 OGD/R 损伤发挥神经保护作用。