缺氧条件下谷氨酸转运体和γ-氨基丁酸转运体在星形胶质细胞中的表达变化及作用  

Expressions of glutamate and GABA transporters in astrocytes under hypoxia and their functions

作　　者：王之涵[1] 邢进[1] 魏子龙[1] 沙龙贵[1] 周新军[1] 林盈盈[2] 邱永明[2] 任力[1] WANG Zhi-han;XING Jin;Wei Zi-long;SHA Long-gui;LIN Ying-ying;QIU Yong-ming;REN Li(Department ofNeurosurgery,Shanghai Pudong Hospital,Fudan University,Shanghai 201300,China;Department of Neurosurgery,Renji Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 2001274,China)

机构地区：[1]复旦大学附属上海市浦东医院神经外科,上海201300 [2]上海交通大学医学院附属仁济医院神经外科,上海200127

出　　处：《中国临床神经外科杂志》2018年第11期735-738,共4页Chinese Journal of Clinical Neurosurgery

基　　金：上海市浦东新区卫生和计划生育委员会重点专科(PWZzk2017-16)

摘　　要：目的探讨缺氧条件下谷氨酸转运体(GLT1)和γ-氨基丁酸转运体(GAT1)在星形胶质细胞中的表达变化及作用。方法星形胶质细胞在含有10%胎牛血清DMEM中培养,在培养液中加入100μmol/L氯化钴溶液处理16、24、48 h模拟缺氧环境。通过慢病毒介导的表达特异性的短发夹RNA(shRNA)沉默GLT1和GAT1基因表达,或者使GLT1和GAT1基因过表达。PCR检测GLT1和GAT1 mRNA表达水平,免疫印迹法检测GLT1和GAT1蛋白表达水平,流式细胞术检测细胞凋亡。结果加入氯化钴处理后,星形胶质细胞GLT1 mRNA和蛋白表达水平显著增高(P<0.05),而GAT1 mRNA和蛋白表达水平显著下降(P<0.05);两者均呈时间依赖性。沉默GLT1基因表达同时过表达GAT1,星形胶质细胞凋亡率明显增加(P<0.05);而过表达GLT1同时沉默GAT1基因表达,星形胶质细胞凋亡率明显减少(P<0.05)。结论缺氧条件下,体外培养的星形胶质细胞GLT1表达上调,而GAT1表达下调,促进星形胶质细胞凋亡。Objective To investigate the expressions and functions of glutamate and γ- aminobutyric acid （GABA） transporters in astrocytes under hypoxic condition. Methods The expressions of glutamate transporter 1 （GLT1） mRNA and protein and GABA transporter 1 （GAT1） mRNA and protein in the astrocytes, who were cultured under the hypoxic condition and received the transfection of GLT1 and GAT1, were detected respectively by quantitetive RT-PCR and west blot. The apoptosis of the astrocytes in whom the expressions of these transporters were changed was determined by flow cytometry. Results Under the hypoxic condition, the expressions of GLT1 and GAT1 mRNA and proteins in the astrocytes was significantly reduced （P〈0.05）. Apoptosis analysis showed that under the hypoxia, inhibition of GLT1 and overexpression of GAT1 significantly increased the astrocytes apoptosis （P〈0.01）, while overexpression of GLT1 and inhibition of GAT1 expression significantly reduced the astrocytes apoptosis （P〈0.01）. Conclusions The present study revealed the expressions and functions of glutamate and GABA transporters in the astrocytes under the ischemia. It is suggested that the increasing in GLT1 expression and decrease in GAT1 expression may have the curative effects on epilepsy or ischemia.

关 键 词：脑缺血 缺氧 星形胶质细胞 细胞凋亡 谷氨酸转运蛋白1 γ-氨基丁酸转运蛋白1 基因表达 

分 类 号：R743[医药卫生—神经病学与精神病学] R363[医药卫生—临床医学]