增强缝隙连接耦联对鱼藤酮染毒星形胶质细胞谷氨酸代谢的影响

Effects of Enhancing the Gap Junction on Metabolism of Glutamate in Rotenone Exposed Astrocytes

目的观察增强缝隙连接耦联对鱼藤酮染毒星形胶质细胞谷氨酸代谢的影响。方法体外培养大鼠中脑星形胶质细胞,随机分为对照组、鱼藤酮组(1.0μmol/L)、缝隙链接激动剂(AAP-10)干预组(50 nmol/L)和AAP-10干预组(250 nmol/L)。MTT法观察染毒星形胶质细胞活力,应用HPLC荧光法和同位素标记法检测星形胶质细胞外谷氨酸浓度和谷氨酸转运能力的变化,RT-PCR检测星形胶质细胞谷氨酸转运体(GLAST)mRNA的表达。结果鱼藤酮染毒组星形胶质细胞活力和谷氨酸转运能力明显降低,细胞外谷氨酸浓度明显增加;AAP-10干预组(250 nmol/L)星形胶质细胞活力有所升高,细胞外谷氨酸浓度明显降低,谷氨酸转运功能活性则明显增强,且GLAST mRNA表达明显上调。结论增强缝隙连接耦联能明显改善鱼藤酮所致星形胶质细胞外谷氨酸代谢紊乱,可能与其调控GLAST基因表达和功能活性有关。

Objective To evaluate the effects of anti-arrhythmic peptide 10（AAP-10）and activator of gap junction on metabolism of glutamate in rotenone exposed astrocytes.Methods MTT assay was used to evaluate the toxicity of rotenone on astrocytes,the level of extracellular glutamate was determined by HPLC,the uptake of glutamate was detected with labeled isotope,and the expression of GLAST mRNA was observed by RT-PCR.Results Compared with 1.0 μmol/L rotenone intoxication group,the viabilities of cell culture were significantly increased and the levels of extracellular glutamate were obviously decreased.The function of glutamate uptake and the expression of GLAST mRNA were enhanced in AAP-10 group（250 nmol/L）.Conclusions Enhancement of gap junction could significantly ameliorate the disorder of glutamate metabolism in astrocytes induced by rotenone,which might be related to the change of GLAST.