Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, P.R. China.
Department of Neurosurgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai 201399, P.R. China.
Mol Med Rep. 2018 Jun;17(6):8196-8202. doi: 10.3892/mmr.2018.8888. Epub 2018 Apr 16.
Glutamate and γ‑aminobutyric acid (GABA) transporters serve central roles in normal neuronal activity and are associated with numerous pathological brain conditions, including ischemia and epilepsy. However, the interplay between these transporters in ischemia remains unclear. In the present study, the expression levels of the excitatory amino acid carrier 1 (EAAC1) and GABA transporter 1 (GAT1) were analyzed in vivo and in vitro within ischemic models by immunofluorescence, western blot and RT‑qPCR. Cell survival rates were analyzed following altered expression of these transporters within neuronal cells by flow cytometry. Expression levels of EAAC1 were reduced within the cerebrum of focal cerebral ischemic middle cerebral artery occlusion rat models as well as in primary neurons cultured under hypoxia. However, GAT1 expression levels were slightly elevated under ischemic conditions. The altered expression levels of EAAC1 and GAT1 were combined within neuron cells and the effects were investigated. Apoptotic analysis revealed that EAAC1 suppression and overexpression of GAT1 increased neuronal cell apoptosis under hypoxic conditions; however, EAAC1 overexpression combined with GAT1 knockdown reduced neuronal cell apoptosis under hypoxic conditions. The present study detected the expression levels of the glutamate and GABA transporters under hypoxia, in association with ischemia. The results indicated that, increased expression of EAAC1 combined with GAT1 suppression may provide protective effects in the treatment of epilepsy and ischemia.
谷氨酸和γ-氨基丁酸（GABA）转运体在正常神经元活动中起着核心作用，与许多病理性脑状况有关，包括缺血和癫痫。然而，在缺血中这些转运体之间的相互作用仍不清楚。在本研究中，通过免疫荧光、Western blot 和 RT-qPCR 分析了兴奋性氨基酸载体 1（EAAC1）和 GABA 转运体 1（GAT1）在体内和体外缺血模型中的表达水平。通过流式细胞术分析了这些转运体在神经元细胞中的表达改变后细胞存活率。在局灶性大脑中动脉闭塞大鼠脑缺血模型的大脑中以及在缺氧培养的原代神经元中，EAAC1 的表达水平降低。然而，在缺血条件下，GAT1 的表达水平略有升高。在神经元细胞中改变 EAAC1 和 GAT1 的表达水平并研究其作用。凋亡分析显示，在缺氧条件下，EAAC1 抑制和 GAT1 过表达增加神经元细胞凋亡；然而，EAAC1 过表达与 GAT1 敲低结合可减少缺氧条件下神经元细胞的凋亡。本研究检测了缺氧和缺血条件下谷氨酸和 GABA 转运体的表达水平。结果表明，EAAC1 的表达增加与 GAT1 的抑制结合可能为治疗癫痫和缺血提供保护作用。
