摘要
目的:比较欧洲越桔和笃斯越桔对氧化应激损伤PC12细胞的保护作用,并观察其作用是否具有剂量依赖性。方法:实验于2004-10/2005-01在华东理工大学生物工程学院生物反应器国家重点实验室进行。PC12细胞以含12.5,25,50mg/L的欧洲越桔标准提取物和笃斯越桔标准提取物的DMEM培养液预孵8h后,以500μmol/L的H2O2干预12h作为氧化应激损伤的模型,采用MTT比色法计算细胞死亡抑制率评估细胞存活情况(抑制率越高,存活细胞越多),DCFH-DA荧光染色法测定细胞内活性氧化物清除抑制率评估细胞内活性氧化物水平(抑制率越低,细胞内活性氧化物水平越低),Hoechst染色法观察细胞核形态学,流式细胞仪检测细胞凋亡。结果:①细胞死亡抑制率:笃斯越桔标准提取物25,50mg/L组高于欧洲越桔标准提取物25,50mg/L组(27.78%,23.66%;46.05%,41.67%),两种药物均为剂量越大,其抑制率越高。②PC12细胞内活性氧化物清除抑制率:笃斯越桔标准提取物25,50mg/L组低于欧洲越桔标准提取物25,50mg/L组(67.58%,80.61%;41.86%,56.69%),且剂量越大,其抑制率越低。③细胞凋亡率:正常对照组为0.69%,500μmol/LH2O2作用12h后,为30.19%。预先给予欧洲越桔标准提取物和笃斯越桔标准提取物处理后,分别为9.45%,8.82%。结论:欧洲越桔和笃斯越桔对氧化应激损伤PC12细胞均具有保护作用,笃斯越桔作用优于欧洲越桔,且具有剂量依赖性效应。其机制可能与其能降低细胞内活性氧化物水平,进而抑制神经细胞凋亡有关。
AIM： To compare the protective effect of Vaccinium myrtillus L. and Vaccinium uliginosum L. on PC12 cells damage induced by oxidative stress, and observe the effect with dose-dependent or without dose-dependent. METHODS： The experiment was done in the National Key Laboratory of Bioreactor, Institute of Biological Engineering, East China University of Science and Technology from October 2004 to January 2005. The PC12 cells were pre-incubated in the DMEM medium of the 12.5, 25, 50 mg/L standard extract of Vaccinium myrtillus L. and Vaccinium uliginosum L. for 8 hours, and intervened for 12 hours with 500μmol/L H202 as the injured model induced by oxidative stress. The dead inhibitive rate of cells was calculated with the MTT chromatometry to evaluate the living condition of cells （the higher the inhibitive rate was, the more the living cells had）. The inhibitive rate of intracellular reactive oxygen species （ROS） was assessed with DCFH-DA fluorescent staining to assess the level of intracellular active oxide （the lower the inhibitive rate was, the lower the level of intraeellular active oxide was）. The nuclear morphology was observed with Hoechst staining. The apoptosis was detected with the flow cytometer. RESULTS： ① Cell dead inhibitive rate： It was higher in the 25,50 mg/L standard extract of Vaccinium uliginosum L. group than that in the 25,50 mg/L standard extract of Vaccinium myrtillus L. group （ 27.780/0, 23.66%; 46.05%, 41.67%）, and the bigger the dosage of the two kinds of drug was, the higher the inhibitive rate was.② Cleaning inhibitive rate of active oxide in PC12 cells： It was lower in the 25,50 mg/L. standard extract of Vaccinium uliginosum L. group than that in the 25,50 mg/L standard extract of Vaccinium myrtillus L. （67.58%, 80.61%; 41.86%, 56.69%）, and the bigger the dosage was, the lower the inhibitive rate was. ③ Apoptosis rate： It was 0.69% in the normal control group, and after 12 hours action of 500μmol/L H2O2 it was 30.19%. After the disposal of standard extract of Vaccinium myrtillus Land Vaccinium uliginosum L., it was 9.45%, 8.82%, respectively. CONCLUSION： The Vaccinium myrtillus L. and Vaccinium uliginosum L. have protective effects on the PC12 ceils damage＇induced by oxidative stress, and the effect of Vaccinium uliginosum L. is better than that of the Vaccinium myrtillus L., and with dose-dependent effects. The mechanism may have relation with its decrease the level of active oxide in ceils so as to inhibit the neural apoptosis.
出处
《中国临床康复》
CSCD
北大核心
2005年第33期50-52,i0002,共4页
Chinese Journal of Clinical Rehabilitation
作者简介
陶欣艺★，女，1976年生，江苏省盐城市人，汉族，2004年南京中医药大学毕业，硕士，主要从事药理学研究工作。Taoxy066@126．com
