不同低密度脂蛋白氧化模型中维生素E的抗氧化效率差异  

Antioxidative Efficiency of Vitamin E in Different LDL Oxidation Models of Atherosclerosis

作　　者：周莉娜[1] 吴露[1] 许程洁[2] 项明伟 宋健[1] 武军驻[1] 

机构地区：[1]武汉大学基础医学院,湖北省武汉市430071 [2]四川省医学科学院 四川省人民医院检验科,四川省成都市610072

出　　处：《中国动脉硬化杂志》2014年第7期663-668,共6页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金(81170273);国家大学生创新性实验计划项目(201310486080);武汉大学本科教学开放实验项目

摘　　要：目的探讨维生素E(VE)在不同低密度脂蛋白(LDL)氧化模型中抗氧化效率的差异,并揭示VE在体外抗氧化效率不高的可能原因。方法分别建立LDL主动氧化模型和被动氧化模型,观察LDL与VE共刺激0、12h以及脂多糖(LPS)与VE共刺激0、3 h后细胞内人α防御素(HNP-1)蛋白及其mRNA水平的表达变化,再通过检测羟基化蛋白(PCO)、丙二醛(MDA)来判断VE是否干扰模型的建立。再将两种氧化模型分别设立氧化模型建立前、后加入VE以及VE(-)组,最后检测不同时间点加入VE后MDA、PCO的变化以分析VE在不同模型中的抗氧化效率差异,并观察细胞内氧自由基水平的波动以判断差异产生的可能原因。结果 (1)氧化模型中HNP-1mRNA及其蛋白水平的表达均升高,MDA和PCO的生成量也显著升高(P<0.05)。(2)主动氧化模型中不同时间点加入VE后PCO生成量无明显差异(P>0.05);被动氧化前加VE组氧化产物生成量明显低于被动氧化后加VE组(P<0.05)。不同氧化模型的MDA、PCO生成量在氧化前加VE组和氧化后加VE组降低,且被动氧化模型中下降幅度更加明显,而VE(-)组含量变化趋势相同。(3)不同氧化模型中加入VE,均能够显著提高超氧化物歧化酶(SOD)活性;能够降低自由基水平,且在被动氧化前加VE组自由基水平降低更明显(P>0.05)。结论 VE的抗氧化效率在被动氧化模型中优于主动氧化模型,与VE的加入时间、胞内自由基水平相关。Aim To detect the anti-oxidation efficiency of vitamin E in different low density lipoprotein（LDL）oxidation models and explore the possible reasons for low antioxidant efficiency of vitamin E. Methods The active LDL oxidation model and passive LDL oxidation model were established in CLR-1730 cell line respectively. After the costimulation with LDL and VE for 0 h,12 h,and co-stimulation with LPS and VE for 0 h,3 h,the mRNA and protein expression levels of human alpha-defensin-1（HNP-1）were detected by using real time PCR and ELISA. We set up 3 different VE working groups： VE added before the oxidation model（pre-VE）,vitamin E added after oxidation model（post-VE）and the control[VE（-）]. We detected malondialde（ MDA）,protein carbonyl（ PCO） and super oxide dlsmutase（SOD） to determine whether vitamin E interfere with the oxidation model. The level of oxygen free radicals in cells were observed by using flow cytometry and fluorescence microscopy. Results（1） The mRNA levels and protein levels of HNP-1 were increased in oxidation models（ P 〈 0. 05）.（2） PCO detection among the three groups in active model showed no significant difference（P 〉 0. 05）; while in passive model the pre-VE and post-VE results were significantly lower than the VE（-）（P 〈 0. 05）. PCO detection： the difference during pre-VE and post-VE in active model was not obvious,but both were significantly lower than the VE（-）（P 〈 0. 05）. SOD detection： pre-VE in active model was significantly higher than post-VE and VE（-）（P 〈 0. 05）.（3） In both oxidation models,the intracellular oxygen free radicals in the pre-VE and post-VE groups were higher than the control. Conclusion VE would not affect the establishment of active and passive oxidation models. The antioxidant efficiency of anti-lipid and protein oxidation is significantly different,while the activity of SOD were enhanced in both models.

关 键 词：主动氧化 被动氧化 低密度脂蛋白 脂多糖 氧自由基 超氧化物歧化酶 

分 类 号：R363[医药卫生—病理学]