非洛地平对氧化损伤的人脐静脉内皮细胞的保护作用

Protective effect of felodipine on human umbilical vein endothelial cells injured by oxidized low-density lipoprotein

目的研究非洛地平(felodipine)对氧化型低密度脂蛋白(ox-LDL)损伤的人脐静脉内皮细胞(HUVECs)活性氧(ROS)及细胞间粘附分子-1(ICAM-1)、血管细胞粘附分子-1(VCAM-1)mRNA表达的影响,探讨其独立于降血压外的抗动脉粥样硬化的作用机制。方法筛选ox-LDL与HUVECs细胞孵育最适的干预浓度梯度与时间,然后与不同浓度的非洛地平(0.1、1、10μmol/L)共同孵育24h,流式细胞仪测定细胞内ROS,realtime-PCR检测细胞ICAM-1、VCAM-1 mRNA的表达。结果 ox-LDL对HUVECs内ROS的产生呈浓度依赖效应,随着刺激浓度的升高,ROS产生增多,25mg/Lox-LDL作用最为显著(P<0.01),非洛地平可抑制ox-LDL诱导的HUVECsROS(P<0.05)的产生;随ox-LDL刺激浓度的增加,ICAM-1、VCAM-1 mRNA表达逐渐上调,当浓度达到25mg/L时,作用最为显著(P<0.05),非洛地平可下调ICAM-1、VCAM-1 mRNA的表达(P<0.05)。结论非洛地平可抑制ox-LDL诱导的HUVECs ROS的产生以及下调ICAM-1、VCAM-1 mRNA表达,发挥抗氧化抗炎的内皮保护功能。

Objective To investigate the protective effect of felodipine on reactive oxygen species （ROS） generation,mRNA level of inflammatory factors such as intercellular adhesion molecule-1 （ICAM-1） and vascular cell adhesion molecule-1 （VCAM-1）,in human umbilical vein endothelial cells （HUVECs） injured by oxidized low-density lipoprotein （ox-LDL） so as to explore felodipine＇s anti-atherosclerosis mechanism independent of its anti-hypertensive effect. Methods Isolated HUVECs were treated with ox-LDL at different concentrations （6,12.5 and 25 mg/L） for 24 hours so that the optimal concentration and time of ox-LDL treatment were selected. Then the cells were incubated with ox-LDL and treated with felodipine at different concentrations （0.1,1 and 10 μmol/L）. Intracellular ROS level was determined by flow cytometry （FCM）. Expressions of inflammatory factors ICAM-1 and VCAM-1 were measured by real time-polymerase chain reaction （real time-PCR）. Results ROS generation was increased in HUVECs after treatment with different concentrations （6,12.5 and 25 mg/L） of ox-LDL for 24 hours and there was a significant difference at 25 mg/L ox-LDL （P0.01）; ROS generation was decreased after felodipine treatment for 24 hours （P0.05）. The mRNA expression of ICAM-1 and VCAM-1 in HUVECs during treatment with 25 mg/L ox-LDL was significantly higher than that in the control group （P0.05）,but ICAM-1 mRNA and VCAM-1 mRNA expressions were significantly down-regulated during treatment with 0.1 μmol/L felodipine （P0.05）. Conclusion Felodipine has an anti-atherosclerosis endothelial protective effect. The anti-oxidation and anti-inflammation mechanism may be related to its inhibiting HUVECs ROS generation induced by ox-LDL as well as down-regulating ICAM-1 and VCAM-1mRNA expressions.