天麻素和异钩藤碱联合应用对MPP^(+)诱导的PC12细胞ROS产生的协同抑制作用  

The Combination of Gastrodin and Isorhynchophylline Exhibits a Synergistic ROS-scavenging Action in MPP^(+)-challenged PC12 Cells

作　　者：于春磊[1] 李强[1] 牛英才[1] 董妙先[1] YU Chun-lei;LI Qiang;NIU Ying-cai;DONG Miao-xian(Institute of Medicine,Qiqihar Medical College,Qiqihar Heilongjiang 161006,China)

机构地区：[1]齐齐哈尔医学院医药科学研究所,黑龙江齐齐哈尔161006

出　　处：《时珍国医国药》2021年第5期1046-1050,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81873133)。

摘　　要：目的观察天麻素联合异钩藤碱对1-甲基-4-苯基吡啶离子(MPP^(+))诱导的PC12细胞活性氧簇(ROS)产生的影响,并探讨其可能的作用机制。方法采用DCFH-DA探针检测细胞内ROS水平,采用化学发光法测定细胞还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)含量,采用实时定量PCR法检测血红素加氧酶-1(HO-1)、醌氧化还原酶-1(NQO-1)、谷氨酸半胱氨酸连接酶催化亚基(GCLc)、修饰亚基(GCLm)和硫氧还蛋白1(Trx1)mRNA表达,采用CompuSyn软件计算天麻素和异钩藤碱联合应用的联合指数和最佳搭配浓度。结果1μM和0.3μM分别为天麻素和异钩藤碱抑制MPP^(+)诱导的PC12细胞ROS产生的最小有效浓度,24 h为天麻素和异钩藤碱抑制MPP^(+)诱导的PC12细胞ROS产生的最佳孵育时间,1μM天麻素联合0.3μM异钩藤碱清除ROS协同效应最强。天麻素联合异钩藤碱显著增加MPP^(+)处理的PC12细胞总GSH含量和GSH/GSSG比值,上调HO-1、NQO1、GCLc和Trx1 mRNA表达,而天麻素联合异钩藤碱的这些药理作用与调控细胞外调节蛋白激酶(ERK1/2)、蛋白激酶B(Akt)和糖原合成激酶-3β(GSK-3β)信号通路有关。结论天麻素联合异钩藤碱对MPP^(+)诱导的PC12细胞ROS清除具有协同效应,其机制可能与上调HO-1、NQO1、GCLc和Trx1 mRNA表达有关。Objective The aim of this study was to investigate the synergistic ROS-scavenging effect of Gastrodin in combination with Isorhynchophylline in MPP^(+)-challenged PC12 cells,and some mechanisms involved.Methods A dichlorofluorescin assay was used to determine cellular ROS generation.Measurements of GSH and GSSG content were performed using luminescent detection.Real-time PCR was used to analyze the mRNA expressions of HO-1,NQO1,GCLc,GCLm and Trx1.Synergistic action between the Gastrodin and Isorhynchophylline was indicated by the combination index using CompuSyn software.Results The minimum effective concentration of Gastrodin and Isorhynchophylline to scavenge ROS in MPP^(+)-challenged PC12 cells are 1μM and 0.3μM,respectively.24 h is the suitable incubation time of both Gastrodin and Isorhynchophylline.The maximal synergism was found when 0.3μM Isorhynchophylline was combined with 1μM Gastrodin.The combination of Gastrodin and Isorhynchophylline enhanced GSH concentration and GSH/GSSG ratio in MPP^(+)-challenged PC12 cells.The combination of Gastrodin and Isorhynchophylline up-regulated the HO-1,NQO1,GCLc and Trx1 mRNA expressions in MPP^(+)-challenged PC12 cells.ERK1/2,Akt,and GSK-3βsignaling are involved in these changes mediated by the combination of Gastrodin and Isorhynchophylline.Conclusion The combination of Gastrodin and Isorhynchophylline exhibits a synergistic ROS-scavenging effect in MPP^(+)-challenged PC12 cells.Possible mechanisms for the actions of Gastrodin in combination with Isorhynchophylline may be relevant with up-regulation of HO-1,NQO1,GCLc,and Trx1 mRNA expressions.

关 键 词：天麻素 异钩藤碱 氧化应激 联合指数 谷胱甘肽 

分 类 号：R992[医药卫生—毒理学]