雙孔鉀通道TREK-1在缺血星形膠質(zhì)細(xì)胞谷氨酸代謝中的作用
發(fā)布時(shí)間：2018-10-15 17:31
【摘要】：目的：1.研究氧糖剝奪對(duì)星形膠質(zhì)細(xì)胞雙孔鉀通道TREK-1表達(dá)及谷氨酸攝取功能的影響。2.研究星形膠質(zhì)細(xì)胞TREK-1通道表達(dá)變化對(duì)谷氨酸興奮性毒性的調(diào)節(jié)作用。 方法：1.采用經(jīng)典體外腦缺血模型：即細(xì)胞氧糖剝奪模型(OGD),分析體外腦缺血時(shí)星形膠質(zhì)細(xì)胞TREK-1雙孔鉀通道表達(dá)變化及谷氨酸攝取功能的變化。2.分別使用TREK-1激動(dòng)劑花生四烯酸(AA)和抑制劑甲硫氨酸(M),進(jìn)行OGD實(shí)驗(yàn)。用MTT法檢測(cè)OGD在1h、4h、8h的細(xì)胞抑制率,Hoechst33258染色法檢測(cè)細(xì)胞死亡率,流式細(xì)胞儀FITC/Propidium Iodide雙染色法檢測(cè)星形膠質(zhì)細(xì)胞凋亡率。RT-PCR法檢測(cè)星形膠質(zhì)細(xì)胞特異性谷氨酸轉(zhuǎn)運(yùn)體GLT-1mRNA及NR2B-1mRNA、Mglur-1mRNA、Caspase-3mRNA、Trek-1mRNA、Erk-1mRNA等變化。 結(jié)果：1.體外腦缺血時(shí),與正常培養(yǎng)的星形膠質(zhì)細(xì)胞相比較,發(fā)現(xiàn)TREK-1雙孔鉀通道表達(dá)在OGD4h后明顯增加,谷氨酸攝取功能增強(qiáng)。2.分別使用TREK-1激動(dòng)劑花生四烯酸(AA)和抑制劑甲硫氨酸(M),進(jìn)行OGD實(shí)驗(yàn),AA通過激活TREK-1通道,具有降低谷氨酸興奮性毒性的作用,從而增加細(xì)胞存活率。M作用于TREK-1通道,谷氨酸的含量明顯增加,興奮性毒性增強(qiáng)。說(shuō)明星形膠質(zhì)細(xì)胞TREK-1具有降低谷氨酸興奮性毒性作用。 結(jié)論：1.氧糖剝奪時(shí),星形膠質(zhì)細(xì)胞雙孔鉀通道表達(dá)增加,對(duì)谷氨酸的攝取功能增強(qiáng)。2.激活星形膠質(zhì)細(xì)胞雙孔鉀通道TREK-1,可以降低谷氨酸的興奮性毒性作用。
[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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3
本文編號(hào)：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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
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2 李正斌;王曉良;;花生四烯酸敏感和機(jī)械牽張門控的雙孔鉀離子通道TREK的研究進(jìn)展[J];藥學(xué)學(xué)報(bào);2006年03期
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