多巴胺通过mTOR-EAAT2通路影响星形胶质细胞谷氨酸摄取能力

发布时间：2018-04-25 09:28

本文选题：多巴胺 + 谷氨酸　；参考：《中国病理生理杂志》2017年10期

【摘要】：目的:研究多巴胺(dopamine,DA)对星形胶质细胞谷氨酸(glutamate,Glu)摄取能力的影响,以及DA通过哺乳动物雷帕霉素靶蛋白(m TOR)-兴奋性氨基酸转运体2(EAAT2)信号通路对星形胶质细胞Glu摄取能力的影响。方法:采用Amplex Red谷氨酸测定试剂盒检测经过干预的原代皮层星形胶质细胞对Glu摄取含量的变化,RT-q PCR、Western blot和免疫荧光染色等检测EAAT2和m TOR mRNA和蛋白质相对表达量,m TOR拮抗剂雷帕霉素或m TOR兴奋剂MHY1485干预在DA中共培养的星形胶质细胞,检测m TOR和EAAT2的表达情况,以及培养上清液Glu的含量。结果:DA干预的原代星形胶质细胞中m TOR表达下调,EAAT2表达下调,培养上清液Glu水平上升;雷帕霉素干预后,EAAT2表达下调,培养上清液中Glu的含量增加;MHY1485干预后,EAAT2表达上调,培养上清液中Glu的含量下降。结论:DA通过与星形胶质细胞m TOR-EAAT2通路相互作用,减弱星形胶质细胞摄取Glu的能力,引起细胞外Glu蓄积,最终损伤星形胶质细胞的功能。
[Abstract]:Aim: to study the effect of dopamine dopamine (DA) on the glutamate glutamate (Glu) uptake in astrocytes and the effect of DA on the Glu uptake of astrocytes through the mammalian Rapamycin target protein (ORM) -excitatory amino acid transporter (2EAAT2) signaling pathway. Methods: the changes of Glu uptake by primary cortical astrocytes were detected by Amplex Red assay kit. The relative expression of EAAT2 and m TOR mRNA and protein were detected by RT-PCR blot and immunofluorescence staining. The antagonist rapamycin or m TOR stimulant MHY1485 interfered with astrocytes co-cultured in DA. The expression of m TOR and EAAT2 and the content of Glu in culture supernatant were detected. Results the expression of m TOR was down-regulated and the level of Glu in culture supernatant was increased in primary astrocytes treated with 1: DA, and the expression of Glu in culture supernatant was increased after intervention of rapamycin, and the expression of Glu in culture supernatant increased after intervention by MHY1485, and the expression of EAAT2 increased after intervention by rapamycin. The content of Glu in culture supernatant decreased. Conclusion by interacting with the m TOR-EAAT2 pathway of astrocytes, the ability of Glu uptake by astrocytes is weakened, and the accumulation of extracellular Glu is induced and the function of astrocytes is damaged.
【作者单位】：温州医科大学附属第一医院浙江省神经老化与疾病研究重点实验室;
【基金】：国家自然科学基金资助项目(No.81671042;No.81300308)
【分类号】：R741

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