羟基红花黄色素A对过氧化氢致L02细胞谷胱甘肽氧化的影响  

Effect of Hydroxysafflor Yellow A on the Oxidation of Intracellular Glutathione Induced by Hydrogen Peroxide in L02 Cells

作　　者：王晓娜[1] 徐晓敏[1] 邱理红 李刚[2] 许波[2] 马成俊[2] 王振华[1,2] 

机构地区：[1]新疆特种植物药资源教育部重点实验室,石河子大学药学院,新疆石河子832002 [2]烟台大学生命科学学院,山东烟台264005

出　　处：《食品科学》2013年第23期317-320,共4页Food Science

基　　金：国家自然科学基金项目(11175222);教育部"新世纪优秀人才支持计划"项目(NCET-10-0967);新疆生产建设兵团医药专项项目(2011BA039);山东省自然科学基金项目(ZR2009BM027);烟台市科技发展计划项目(201025)

摘　　要：研究羟基红花黄色素A（HSYA）对过氧化氢（H2O2）所致L02人胎肝细胞内谷胱甘肽氧化的影响。采用5,5’-二硫代双硝基苯甲酸（DTNB）衍生化法,用高效液相色谱（HPLC）法测定L02细胞内还原型谷胱甘肽（GSH）浓度,二硫苏糖醇（DTT）还原细胞内氧化型谷胱甘肽（GSSG）后测定细胞内总GSH含量,以此计算出细胞内GSSG含量。HSYA预处理L02人胎肝细胞24h,再加入H2O2孵育1h后,利用HPLC测定细胞内GSH和GSSG水平及GSSG/GSH比值的变化。结果表明：GSH在0.1-2mmol/L范围内线性关系良好,最低定量限为0.1mmol/L,批内、批间精密度均小于10%,样品的回收率均在95%-105%之间。此测定GSH方法准确、精密、重复性好、稳定性高,可用于细胞中GSH和GSSG的测定。H2O2处理后L02细胞内GSSG/GSH比值明显升高,HSYA能够降低L02细胞中GSSG水平,使GSSG/GSH的比值下降,表明HSYA能够改善H2O2所致L02细胞的氧化胁迫状态,保护细胞免受氧化损伤。The effect of hydroxysafflor A （HSYA） on the oxidation of intracellular reduced glutathione （GSH） induced by hydrogen peroxide （H2O2） was investigated in L02 ceils. 5,5＇-Dithiobis（2-nitrobenzoic acid） （DTNB） was used as the derivatizing agent and the reduced glutathione was measured by HPLC. The total glutathione was measured by the same method after reduced by dithiothreitol （DTT）. Oxidative glutathione can be calculated through reduced glutathione and total glutathione. The L02 cells were pretreated with HSYA for 24 hours, and then exposed to H2O2 for 1 hour. The intracellular GSH, GSSG contents and GSSG/GSH were analyzed by HPLC. The results showed that the standard curve was linear over the range of 0.1-2 mmol/L. The minimum quantification limit was 0.1 mmol/L. The precision discrepancy of inter-batch and intra-batch assays were both less than 10%. The recovery rates at low, medium and high spiked concentrations were in the range of 95%- 105%. The developed method proved to accurate, quick, precise and suitable for the measurement of glutathione in cells. HSYA can significantly inhibit the increase of GSSG and GSSG/GSH, thus relieving the oxidative stress induced by H2O2.

关 键 词：羟基红花黄色素A 谷胱甘肽 高效液相色谱 氧化胁迫 

分 类 号：R962[医药卫生—药理学]