机构地区:[1]青岛大学医学院附属医院心内科,山东青岛266003 [2]青岛大学医学院药理学教研室,山东青岛266071 [3]青岛大学医学院附属医院急诊科,山东青岛266003 [4]青岛大学医学院附属医院中心实验室,山东青岛266071
出 处:《现代生物医学进展》2014年第11期2019-2023,共5页Progress in Modern Biomedicine
基 金:Natural Science Foundation of China(81173593)~~
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1.5μmlo/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4.5μmlo/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。Objective: To investigate the effects of Ursolic acid on NQO1 (Quinone Oxidoreductasel, 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): Group Ⅰ, control group without; Group Ⅱ, HUVECs stimulated with ox-LDL (20 mg/L) in endothelial basal medium for 24 hours; GrouplⅢ, HUVECs treated with ox-LDL(20m g/L) for an half hours, and treated with ox-LDL (20 mg/L) and UA (1.5 μmol/L) in endothelial basal medium for 24 hours; GrouplV, HUVECs treated with ox-LDL(20 mg/L) for an half hours, and treated with ox-LDL (20 mg/L) and UA (4.5μmol/L) in endothelial basal medium for 24 hours. MTT assay was used to determine against HUVECs injury. Expression of NQOlmRNA 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 NQOlmRNA was significantly highter in group Ⅱ (0.624± 0.009)than that in group Ⅰ (0.521± 0.007), P〈0.01. Ursolic acid dose-dependenly increased expression of NQO1 mRNA (GroupⅢvs Group Ⅱ : 0.722:1:0.058 vs 0.624± 0.009, P〈0.01; Group IV vs Group Ⅱ : 0.826 ± 0.059 vs 0.624 ± 0.009, P〈0.01). Expression of NQO1 Protein was significantly highter in group Ⅱ (0.574± 0.024)than that in group Ⅰ (0.438± 0.039), P〈0.01. Ursolic acid dose-dependenly increased expression of NQO1 Protein(Group Ⅲvs Group Ⅱ : 0.710± 0.058 vs 0.574± 0.024, P〈0.01; Group Ⅳ vs Group Ⅱ : 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 re-ducing ox-LDL-induced oxidative stress responses.
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
分 类 号:R543[医药卫生—心血管疾病]
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