文章摘要
苏元镇 王春波 辛辉 张蕊 田嘉伟 张进松 姚如永.熊果酸对ox-LDL诱导的人脐静脉血管内皮细胞NQO1 表达的影响[J].,2014,14(11):2019-2023
熊果酸对ox-LDL诱导的人脐静脉血管内皮细胞NQO1 表达的影响
Effect of Ursolic Acid on NQO1 Expression in HUVECsStimulated by ox-LDL
  
DOI:
中文关键词: 熊果酸  氧化性低密度脂蛋白  醌还原氧化酶1  氧化应激  动脉粥样硬化
英文关键词: Ursolic acid  Ox- LDL  NQO1  Oxidative stress  Atherosclerosis
基金项目:国家自然科学基金项目(81173593)
作者单位
苏元镇 王春波 辛辉 张蕊 田嘉伟 张进松 姚如永 青岛大学医学院附属医院心内科
青岛大学医学院药理学教研室
青岛大学医学院附属医院急诊科
青岛大学医学院附属医院中心实验室 
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中文摘要:
      目的:研究熊果酸对经氧化性低密度脂蛋白(ox-LDL)干预后人脐静脉血管内皮细胞(human umbilical vein endothelial cells, HUVECs)醌还原氧化酶1 表达的影响,以进一步探讨熊果酸抗动脉粥样硬化的机制。方法:体外培养人脐静脉内皮细胞,进行分 组处理,每组n=5。对照组,不加任何处理;ox-LDL组,加入ox-LDL 培养24h,终浓度为20mg/L;ox-LDL+ 低浓度熊果酸组,先加 入ox-LDL(浓度20mg/L)孕育半小时,然后与熊果酸(浓度1.5umlo/L)共同培养24h;ox-LDL+ 高浓度熊果酸组,先加入ox-LDL (浓度20mg/L)孕育半小时,然后与熊果酸(浓度4.5滋mlo/L)共同培养24h;采用MTT 试验测定细胞吸光度值,检测熊果酸对 ox-LDL损伤的保护作用,采用RT-PCR 法检测NQO1mRNA的表达,采用Western blot 法检测NQO1蛋白的表达。结果:熊果酸 减弱ox-LDL对HUVECs的损伤作用;ox-LDL 组NQO1mRNA 的表达量(0.624± 0.009)明显高于对照组(0.521± 0.007),P<0.01。熊 果酸呈浓度依赖性的提高NQO1mRNA的表达量(ox-LDL+ 低浓度熊果酸组vs ox-LDL 组:0.722± 0.058 vs 0.624± 0.009, P<0. 01;ox-LDL+ 高浓度熊果酸组vs ox-LDL 组:0.826± 0.059 vs 0.624± 0.009, P<0.01)。ox-LDL 组NQO1 蛋白的表达量(0.624± 0.009)明显高于对照组(0.521± 0.007),P<0.01。熊果酸呈浓度依赖性的提高NQO1 蛋白的表达量(ox-LDL+ 低浓度熊果酸组vs ox-LDL 组:0.710± 0.058 vs 0.574± 0.024, P<0.01;ox-LDL+ 高浓度熊果酸组vs ox-LDL 组:0.831± 0.034 vs 0.574± 0.024, P<0. 01)。结论:熊果酸可上调ox-LDL诱导的人脐静脉血管内皮细胞NQO1 的表达,表明其可能具有抗氧化应激及抗动脉粥样硬化的 作用。
英文摘要:
      Objective: To investigate the effects ofUrsolic acid on NQO1 (Quinone Oxidoreductase1, NQO1)incultured umbilical vein endothelial cells (HUVECs) stimulated by ox-LDL, so as to approach the mechanism of UA in atherosclerosis.Methods: HUVECs were cultured in different concentration and divided into four groups(n=5,each): GroupⅠ, control group without; GroupⅡ, HUVECs stimulated with ox-LDL (20 mg/L) in endothelial basal medium for 24 hours; GroupⅢ, HUVECs treated with ox- LDL(20m g/L) for an half hours, and treated with ox- LDL (20 mg/L) and UA (1.5 滋mol/L) in endothelial basal medium for 24 hours; GroupⅣ , HUVECs treated with ox- LDL(20 mg/L) for an half hours, and treated with ox- LDL (20 mg/L) and UA (4.5umol/L) in endothelial basal medium for 24 hours. MTT assay was used to determine against HUVECs injury. Expression of NQO1mRNA was determined by RT-PCR. Expression of NQO1 protein was determined by Western blot.Results:Ursolic acid decreased the effect on reducing the cytotoxicity of ox- LDL. Expression of NQO1mRNA was significantly highter in groupⅡ(0.624± 0.009)than that in groupⅠ(0.521± 0.007), P<0.01. Ursolic acid dose-dependenly increased expression of NQO1 mRNA (GroupⅢ vs GroupⅡ : 0.722± 0.058 vs 0.624± 0.009, P<0.01; GroupⅣ vs GroupⅡ : 0.826 ± 0.059 vs 0.624 ± 0.009, P<0.01). Expression of NQO1 Protein was significantly highter in groupⅡ (0.574± 0.024)than that in groupⅠ(0.438± 0.039), P<0.01. Ursolic acid dose-dependenly increased expression of NQO1 Protein(Group Ⅲ vs GroupⅡ : 0.710± 0.058 vs 0.574± 0.024, P<0.01; GroupⅣ vs GroupⅡ : 0.831± 0.034 vs 0.574± 0.024, P<0.01). Conclusion:NQO1 expression in ox-LDL- treated HUVECs could be increased by UA which suggests that UA may attenuate atherosclerosis by reducing ox-LDL-induced oxidative stress responses.
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