桃叶珊瑚苷对谷氨酸诱导的兴奋性神经毒性的抑制作用研究  

Study on the Inhibitory Effect of Aucubin on Glutamate-Induced Excitatory Neurotoxicity

作　　者：卢仁睿 张莉[1] 王慧慧 李孟[1] 郑晓珂[1,2] 冯卫生 LU Renrui;ZHANG Li;WANG Huihui;LI Meng;ZHENG Xiaoke;FENG Weisheng(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Engineering Technology Research Center for Chinese Medicine Development,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,郑州450046 [2]河南省中药开发工程技术研究中心,郑州450046

出　　处：《中国现代应用药学》2022年第24期3197-3203,共7页Chinese Journal of Modern Applied Pharmacy

基　　金：国家重点研发计划中医药现代化研究重点专项(2017YFC1702800);河南省重大科技专项项目(171100310500);河南省高层次人才特殊支持计划“中原千人计划”-中原领军人才(ZYQR201810080);河南省教育厅河南省科技攻关项目(212102311106);河南省中医药科学研究专项课题(20-21ZY2151)。

摘　　要：目的 研究桃叶珊瑚苷对谷氨酸诱导的兴奋性神经毒性的抑制作用。方法 采用20 mmol·L^(-1)谷氨酸诱导PC-12细胞损伤建立神经毒性模型,加入1,5,10,20,40μmol·L^(-1)的桃叶珊瑚苷处理24 h后,通过MTT法检测细胞活力,用流式细胞术检测细胞内Ca^(2+)和活性氧(reactive oxygen species,ROS)水平,采用In Cell Western检测谷氨酸受体N-甲基-D-天冬氨酸受体(N-methyl-D-aspartic acid receptor,NMDA)亚基NMDAR1的水平,采用ELISA试剂盒检测细胞上清中超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、乳酸脱氢酶(lactate dehydrogenase,LDH)水平。结果 谷氨酸20 mmol·L^(-1)作用24 h可使体外培养的PC-12细胞活力明显下降(P<0.01),细胞内Ca^(2+)、NMDAR1蛋白水平和ROS、MDA、LDH水平明显增加(P<0.01),SOD水平明显降低。10μmol·L^(-1)的桃叶珊瑚苷可以明显提高谷氨酸诱导后PC-12细胞的细胞活力(P<0.01),降低细胞内Ca^(2+)、NMDAR1蛋白水平和ROS、LDH水平(P<0.01),升高细胞内SOD水平(P<0.01)。结论 桃叶珊瑚苷可能通过抑制NMDAR1的表达,从而降低细胞内Ca^(2+)的堆积,并抑制氧化应激水平来改善谷氨酸诱导的PC-12细胞损伤,抑制谷氨酸诱导的兴奋性神经毒性。OBJECTIVE To investigate the inhibitory effect of aucubin on glutamate-induced excitatory neurotoxicity.METHODS A neurotoxicity model was established using 20 mmol·L^(-1)glutamate to induce damage in PC-12 cells.After treatment with 1,5,10,20 and 40μmol·L^(-1)aucubin for 24 h,cell viability was measured by MTT,intracellular Ca^(2+)and reactive oxygen species(ROS) levels were measured by flow cytometry,the level of glutamate receptor N-methyl-D-aspartic acid receptor(NMDAR1) were measured by In Cell Western.And superoxide dismutase(SOD),malondialdehyde(MDA),and lactate dehydrogenase(LDH) levels in cell supernatants were measured by using ELISA kits.RESULTS After treated with20 mmol·L^(-1)glutamate for 24 h,the viability of PC-12 cells was significantly reduced(P<0.01),the levels of intracellular Ca^(2+),NMDAR1 ROS,MOA,LDH were significantly increased (P<0.01),the level of SOD was significantly reduced.10μmol·L^(-1)aucubin could significantly increase the cell viability of PC-12 cells injured by glutamate(P<0.01),it could reduce the levels of intracellular Ca^(2+),NMDAR1,ROS and LDH(P<0.01),and elevate intracellular SOD level(P<0.01).CONCLUSION Aucubin may ameliorate glutamate-induced damage in PC-12 cells by inhibiting NMDAR1 expression,thereby reducing the accumulation of intracellular Ca^(2+),and inhibiting oxidative stress levels to suppress glutamate-induced excitatory neurotoxicity.

关 键 词：桃叶珊瑚苷 N-甲基-D-天冬氨酸受体亚基 细胞内Ca^(2+)水平 谷氨酸 PC-12 

分 类 号：R285.5[医药卫生—中药学]