鱼藤酮对PC12细胞谷氨酸含量及其转运体表达的影响

目的观察鱼藤酮对PC12细胞的毒性与谷氨酸递质水平及谷氨酸转运体表达的关系。方法MTT法检测鱼藤酮染毒PC12细胞活力，高效液相色谱法测定染毒细胞胞外Glu含量，RT-PCR及Western blot技术检测谷氨酸转运体EAAC1 mRNA和蛋白表达。结果0．25μmol／L鱼藤酮染毒24h即可引起PC12细胞活力明显降低，LDH渗出量显著增加；0．5μmol／L以上鱼藤酮诱导PC12细胞释放Glu增加，谷氨酸转运体EAAC1基因及蛋白表达均显著降低。结论鱼藤酮引起神经细胞损伤可能与其增加Glu释放和降低谷氨酸转运体的表达有关。

The toxic effects of rotenone on glutamate concentration and glutamate transporter expression in PC12 cells

Objective To study the relationship between neurotoxicity of rotenone and the expression of glutamate transporter. Methods MTT assay was used to evaluate the toxicity of rotenone on PC12 cells viability. Using high performance liquid chromatography （HPLC） to detect the alteration of glutamate in the extracellular fluid of the PC12 cells. The expression of EAAC1 mRNA and protein were measured by RT-PCR and Western blot. Results The viability of cell cultures was obviously decreased after being treated with 0.25 μmol/L rotenone for 24 h. Rotenone was also shown to increase the release of glutamate in PC12 cells and suppress the expression of EAAC1 significantly. Conclusions The neurotoxicity of rotenone is associated with increased release of glutamate in extracellular fluid, which may be induced by declined expression of glutamate transporter.