双孔钾通道TREK-1在缺血星形胶质细胞谷氨酸代谢中的作用
发布时间：2018-10-15 17:31
【摘要】：目的：1.研究氧糖剥夺对星形胶质细胞双孔钾通道TREK-1表达及谷氨酸摄取功能的影响。2.研究星形胶质细胞TREK-1通道表达变化对谷氨酸兴奋性毒性的调节作用。 方法：1.采用经典体外脑缺血模型：即细胞氧糖剥夺模型(OGD),分析体外脑缺血时星形胶质细胞TREK-1双孔钾通道表达变化及谷氨酸摄取功能的变化。2.分别使用TREK-1激动剂花生四烯酸(AA)和抑制剂甲硫氨酸(M),进行OGD实验。用MTT法检测OGD在1h、4h、8h的细胞抑制率,Hoechst33258染色法检测细胞死亡率,流式细胞仪FITC/Propidium Iodide双染色法检测星形胶质细胞凋亡率。RT-PCR法检测星形胶质细胞特异性谷氨酸转运体GLT-1mRNA及NR2B-1mRNA、Mglur-1mRNA、Caspase-3mRNA、Trek-1mRNA、Erk-1mRNA等变化。 结果：1.体外脑缺血时,与正常培养的星形胶质细胞相比较,发现TREK-1双孔钾通道表达在OGD4h后明显增加,谷氨酸摄取功能增强。2.分别使用TREK-1激动剂花生四烯酸(AA)和抑制剂甲硫氨酸(M),进行OGD实验,AA通过激活TREK-1通道,具有降低谷氨酸兴奋性毒性的作用,从而增加细胞存活率。M作用于TREK-1通道,谷氨酸的含量明显增加,兴奋性毒性增强。说明星形胶质细胞TREK-1具有降低谷氨酸兴奋性毒性作用。 结论：1.氧糖剥夺时,星形胶质细胞双孔钾通道表达增加,对谷氨酸的摄取功能增强。2.激活星形胶质细胞双孔钾通道TREK-1,可以降低谷氨酸的兴奋性毒性作用。
[Abstract]:Purpose 1. To study the effects of oxygen glucose deprivation on TREK-1 expression and glutamate uptake in astrocytes. 2. To study the regulatory effect of TREK-1 channel expression on glutamate excitotoxicity in astrocytes. Method 1: 1. A classical model of cerebral ischemia in vitro: (OGD), was used to analyze the changes of TREK-1 bimole potassium channel expression and glutamate uptake function in astrocytes during cerebral ischemia in vitro. 2. The OGD experiment was carried out with TREK-1 agonist arachidonic acid (AA) and methionine (M),. MTT assay was used to detect the cell inhibitory rate of OGD at 4 h, Hoechst33258 staining was used to detect the cell mortality, flow cytometry FITC/Propidium Iodide double staining was used to detect the apoptosis rate of astrocytes, and RT-PCR method was used to detect the changes of the specific glutamate transporter GLT-1mRNA and NR2B-1mRNA,Mglur-1mRNA,Caspase-3mRNA,Trek-1mRNA,Erk-1mRNA. The result is 1: 1. Compared with the normal cultured astrocytes during cerebral ischemia in vitro, the expression of TREK-1 bimodal potassium channels was significantly increased after OGD4h, and the glutamate uptake function was increased. 2. TREK-1 agonist arachidonic acid (AA) and methionine (M),) were used in the OGD experiment. AA could reduce the excitotoxicity of glutamate by activating TREK-1 channel and increase the cell survival rate. The content of glutamate increased significantly and the excitotoxicity increased. These results suggest that astrocyte TREK-1 can reduce the excitatory toxicity of glutamate. Conclusion 1. During oxygen glucose deprivation, the expression of bimodal potassium channels in astrocytes increased and the uptake of glutamate increased. 2. 2. Activation of double-pore potassium channel TREK-1, in astrocytes can reduce the excitotoxicity of glutamate.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3
本文编号：2273279
[Abstract]:Purpose 1. To study the effects of oxygen glucose deprivation on TREK-1 expression and glutamate uptake in astrocytes. 2. To study the regulatory effect of TREK-1 channel expression on glutamate excitotoxicity in astrocytes. Method 1: 1. A classical model of cerebral ischemia in vitro: (OGD), was used to analyze the changes of TREK-1 bimole potassium channel expression and glutamate uptake function in astrocytes during cerebral ischemia in vitro. 2. The OGD experiment was carried out with TREK-1 agonist arachidonic acid (AA) and methionine (M),. MTT assay was used to detect the cell inhibitory rate of OGD at 4 h, Hoechst33258 staining was used to detect the cell mortality, flow cytometry FITC/Propidium Iodide double staining was used to detect the apoptosis rate of astrocytes, and RT-PCR method was used to detect the changes of the specific glutamate transporter GLT-1mRNA and NR2B-1mRNA,Mglur-1mRNA,Caspase-3mRNA,Trek-1mRNA,Erk-1mRNA. The result is 1: 1. Compared with the normal cultured astrocytes during cerebral ischemia in vitro, the expression of TREK-1 bimodal potassium channels was significantly increased after OGD4h, and the glutamate uptake function was increased. 2. TREK-1 agonist arachidonic acid (AA) and methionine (M),) were used in the OGD experiment. AA could reduce the excitotoxicity of glutamate by activating TREK-1 channel and increase the cell survival rate. The content of glutamate increased significantly and the excitotoxicity increased. These results suggest that astrocyte TREK-1 can reduce the excitatory toxicity of glutamate. Conclusion 1. During oxygen glucose deprivation, the expression of bimodal potassium channels in astrocytes increased and the uptake of glutamate increased. 2. 2. Activation of double-pore potassium channel TREK-1, in astrocytes can reduce the excitotoxicity of glutamate.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3
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