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目的研究鱼藤酮对原代培养星形胶质细胞谷氨酸转运功能的影响。方法应用HPLC荧光法检测鱼藤酮染毒星形胶质细胞胞外Glu浓度,同位素标记法检测Glu摄取能力,采用RT-PCR与Western blot技术观察谷氨酸转运体基因及蛋白表达。结果鱼藤酮染毒星形胶质细胞胞外Glu浓度明显升高,Glu摄取能力显著降低,谷氨酸/天冬氨酸转运体(glutamate/aspartate transporter,GLAST)基因和蛋白表达均明显降低,而谷氨酸转运体-1(glutamatetransporter-1,GLT-1)蛋白表达升高。结论鱼藤酮可显著降低星形胶质细胞谷氨酸摄取功能,引起胞外Glu浓度升高;GLAST表达下调可能是鱼藤酮诱导胞外谷氨酸含量增加的主要原因之一,而GLT-1上调可能为神经细胞自我保护机制,以限制谷氨酸的神经毒作用。
Objective To study the effect of rotenone on the glutamate transport function of primary cultured astrocytes. Methods The intracellular Glu concentration of rotenone-treated astrocytes was detected by HPLC fluorescence spectrophotometry. Glu uptake capacity was measured by isotope labeling method. The gene and protein expression of glutamate transporter was observed by RT-PCR and Western blot. Results The extracellular Glu concentration of astrocytes was significantly increased after rotenone exposure, the Glu uptake capacity was significantly decreased, and the gene and protein expression of glutamate / aspartate transporter (GLAST) decreased significantly Glutamate transporter-1 (GLT-1) protein expression increased. Conclusion Rotenone can significantly reduce glutamate uptake by astrocytes and increase the concentration of extracellular Glu. The down-regulation of GLAST may be one of the main reasons for the increase of extracellular glutamate concentration induced by rotenone, while the up-regulation of GLT-1 may be Self-protection mechanism for nerve cells to limit the neurotoxic effect of glutamate.