灯盏乙素对β淀粉样蛋白诱导的神经母细胞瘤细胞谷氨酸/γ氨基丁酸通路的影响  

Effect of scutellarin on Glu/GABA pathway of neuroblastoma cells induced byβ-amyloid protein

作　　者：李晓宇 于燕妮[1] 郭莉莉 LI Xiaoyu;YU Yanni;GUO Lili(Guizhou Medical University,Guiyang 550004,China;不详)

机构地区：[1]贵州医科大学,贵阳550001 [2]贵阳市第一人民医院

出　　处：《山东医药》2020年第26期1-5,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81360174)。

摘　　要：目的观察灯盏乙素(Scu)对β淀粉样蛋白(Aβ)诱导的神经母细胞瘤细胞谷氨酸(Glu)/γ氨基丁酸(GABA)通路的影响,并探讨其作用机制。方法将神经母细胞瘤SH-SY5Y细胞分为对照组、Aβ组、Scu组、Scu+Aβ组。对照组常规培养,不加任何药物;Aβ组加入1μmol/L Aβ作用24 h;Scu组加入10μmol/L Scu作用48 h;Scu+Aβ组先后加入10μmol/L Scu、1μmol/L Aβ各处理24 h。取各组细胞外液,采用生物化学法检测Glu含量,ELISA法检测GABA含量;取各组细胞,采用Western blotting法检测Glu通路转运蛋白兴奋性氨基酸转运体3(EAAT3)、GABA通路转运蛋白γ氨基丁酸转运体1(GAT-1)、Glu通路受体蛋白N-甲基-D-天门冬氨酸受体亚型1(NMDAR1)表达;黄嘌呤氧化酶法检测细胞总超氧化物歧化酶(T-SOD)活力,硫代巴比妥酸法检测细胞丙二醛(MDA)含量。结果与对照组比较,Scu组各指标差异均无统计学意义(P均>0.05);与对照组比较,Aβ组细胞外液Glu含量增加、GABA含量减少,细胞EAAT3、GAT-1、NMDAR1蛋白表达均增加,细胞T-SOD活力降低、MDA含量增加(P均<0.05);与Aβ组比较,Scu+Aβ组细胞外液Glu含量降低、GABA含量增加,细胞EAAT3、GAT-1、NMDAR1蛋白表达均减少,细胞T-SOD活力升高、MDA含量减少(P均<0.05)。结论Scu能够调控Aβ诱导的神经母细胞瘤细胞Glu/GABA通路,抑制Glu兴奋性神经毒性;其机制可能与下调NMDAR1表达,减少Glu通路受体,同时通过抗氧化作用改善线粒体功能障碍,促进EAAT3、GAT-1对Glu、GABA的转运有关。Objective To observe the effect of scutellarin(Scu)on the glutamate(Glu)/γ-aminobutyric acid(GABA)pathway of neuroblastoma cells induced byβ-amyloid(Aβ),and to explore its mechanism of action.Methods Neuroblastoma cells were divided into the control group,Aβgroup(treated with 1μmol/L Aβfor 24 h),Scu group(treated with 10μmol/L Scu for 48 h)and Scu+Aβgroup(treated with 10μmol/L Scu+1μmol/L Aβfor 24 h).The concentration of extracellular Glu in each group was detected by biochemical method,and the concentration of extracellular GABA was detected by ELISA;the expression levels of excitatory amino acid transporter 3(EAAT3),GABA transporter 1(GAT-1)and N-methyl-D-aspartate receptor subtype 1(NMDAR1)were detected by Western blotting;the content of total superoxide dismutase(T-SOD)was measured by xanthine oxidase method,and the content of malondialdehyde(MDA)was measured by thiobarbituric acid method.Results There were no differences in the test items between the control group and the Scu group(all P>0.05).Compared with the control group,the extracellular Glu increased,while the GABA decreased,the expression of EAAT3,GAT-1 and NMDAR1 increased,the cell T-SOD activity decreased,while MDA content increased in the Aβgroup(all P<0.05);compared with the Aβgroup,the extracellular Glu decreased while the GABA content increased,the expression levels of EAAT3,GAT-1,and NMDAR1 all decreased,the cell T-SOD activity increased,while the MDA content decreased in the Scu+Aβgroup(all P<0.05).Conclusions Scu can regulate the Glu/GABA pathway of neuroblastoma cells induced by Aβand inhibit Glu excitotoxicity.The mechanism may be related to down-regulating the expression of NMDAR1,reducing Glu pathway receptors,improving mitochondrial dysfunction through antioxidant effects,and promoting the transport of Glu and GABA by EAAT3 and GAT-1.

关 键 词：灯盏乙素 神经母细胞瘤细胞 阿尔茨海默病 β淀粉样蛋白 谷氨酸 Γ氨基丁酸 兴奋性氨基酸转运体 γ氨基丁酸转运体 N-甲基-D-天门冬氨酸受体 

分 类 号：R741[医药卫生—神经病学与精神病学]