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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 study the effect of ursolic acid on the expression of quinone reductase 1 (HUVECs) in human umbilical vein endothelial cells (HUVECs) induced by oxidized low density lipoprotein (ox-LDL) Mechanisms of acid resistance to atherosclerosis. Methods: Human umbilical vein endothelial cells were cultured in vitro and divided into groups, n = 5 in each group. The control group, without any treatment; ox-LDL group, adding ox-LDL cultured 24h, the final concentration of 20mg / L; ox-LDL + low concentrations of ursolic acid group, first added ox-LDL (concentration 20mg / L) Hour and then incubated with ursolic acid (1.5 μmlo / L) for 24 h. The ox-LDL + UA group was incubated with ox-LDL (20 mg / L) for half an hour before incubated with ursolic acid 4.5μmlo / L) for 24h. MTT assay was used to determine the cell absorbance to detect the protective effect of ursolic acid on ox-LDL. The expression of NQO1 mRNA was detected by RT-PCR and the expression of NQO1 protein by Western blot. Results: Ursolic acid attenuated the injury of HUVECs induced by ox-LDL. The expression of NQO1 mRNA in ox-LDL group (0.624 ± 0.009) was significantly higher than that in control group (0.521 ± 0.007), P <0.01. UA concentration-dependently increased the expression of NQO1mRNA (ox-LDL + low concentrations of ursolic acid vs ox-LDL: 0.722 ± 0.058 vs 0.624 ± 0.009, P <0.01; vs ox-LDL group: 0.826 ± 0.059 vs 0.624 ± 0.009, P <0.01). The expression of NQO1 protein in ox-LDL group (0.624 ± 0.009) was significantly higher than that in control group (0.521 ± 0.007), P <0.01. UA concentration-dependently increased the expression of NQO1 protein (ox-LDL + low concentration of ursolic acid vs ox-LDL group: 0.710 ± 0.058 vs 0.574 ± 0.024, P <0.01; Group vs ox-LDL group: 0.831 ± 0.034 vs 0.574 ± 0.024, P <0.01). Conclusion: UA can up-regulate the expression of NQO1 in human umbilical vein endothelial cells induced by ox-LDL, indicating that it may have anti-oxidative stress and anti-atherosclerosis effects.