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(英文)

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),P0.01。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,P0.01;ox-LDL+高浓度熊果酸组vs ox-LDL组:0.826±0.059 vs 0.624±0.009,P0.01)。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),P0.01。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,P0.01;ox-LDL+高浓度熊果酸组vs ox-LDL组:0.831±0.034 vs 0.574±0.024,P0.01)。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。

Effect of Ursolic Acid on NQO1 Expression in HUVECs Stimulated by ox-LDL

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.