番茄红素对血管内皮细胞损伤的保护作用

摘要

Objective To study the protective effect of lycopene on vascular endothelial cell injury by cigarette smoke extract ( CSE) . Methods CSE was prepared and the human umbilical vein endothelial cells ( HUVECs) were assigned into four groups, cells in control were untreated, and cells in other three groups were treated by 10%CSE, 10%CSE+1. 0 μmol·L-1 lycopene and 1. 0 μmol·L-1 lycopene, respectively. Cell viability was evaluated using MTT assay. The intracelluar reactive oxygen species ( ROS) level was detected by ROS assay kits. Cell cycle and apoptosis were analyzed by flow cytometry. SIRT1 expression was detected by real-time fluorescence quantification PCR ( qRT-PCR) and Western blot. Results Cell viability in 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene or 1. 0 μmol·L-1 lycopene group was (56. 7±5. 1)%,(75. 6±7. 1)% and (95. 5± 9. 7)%, respectively. ROS assay showed that the relative fluorescence intensity in the control was 25. 3±3. 9, however, in 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene group or 1. 0 μmol·L-1 lycopene group were 67. 3±4. 6, 45. 3±3. 9 and 20. 8±2. 9, respectively. 10%CSE could induce G2 arrested and which could be antagonized by 1. 0 μmol·L-1 lycopene. The apoptosis rate in the control, 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene group or 1. 0 μmol·L-1 lycopene group was (6. 2±0. 5)%, (30.8±4.3)%, (18. 3±1. 9)% and (7. 6±0. 4)%, respectively. As shown in qRT-PCT testing, compared with the control, mRNA of SIRT1 in 10%CSE group, 10%CSE+1. 0 μmol·L-1 lycopene group and 1. 0 μmol·L-1 lycopene group was (0. 51± 0. 03) fold, (0. 84±0. 05) fold, and (1. 31±0. 08) fold compared to the control, the data from western blot were consistent with qRT-PCR results. Conclusion Lycopene can prevent HUVECs from injury induced by CSE by upregulation of SIRT1.%目的：研究番茄红素对香烟烟雾提取物致血管内皮细胞损伤的保护作用。方法将人脐周静脉血管内皮细胞( HUVEC )分为4组,对照组不做任何处理,其余3组分别使用10%香烟烟雾提取物( CSE )、10% CSE+1.0μmol·L-1番茄红素及1.0μmol·L-1番茄红素处理。 MTT法分析各组细胞增殖情况,活性氧簇( ROS)检测试剂盒检测各组细胞内ROS水平,流式细胞仪分析各组细胞周期和凋亡率,实时荧光定量PCR( qRT-PCR)、Western blot技术分析各组细胞沉默信息调节蛋白1( SIRT1)表达水平。结果10%CSE 组、10%CSE+1.0μmol · L-1番茄红素组、1.0μmol·L-1番茄红素组细胞存活率分别为(56.7±5.1)%,(75.6±7.1)%和(95.5±9.7)%。 ROS检测发现,对照组、10% CSE组、10%CSE+1.0μmol·L-1番茄红素组和1.0μmol·L-1番茄红素组相对荧光强度分别为25.3±3.9,67.3±4.6,45.3±3.9,20.8±2.9。10%CSE可引起细胞发生G2期阻滞,番茄红素处理可以缓解此效果。对照组、10% CSE组、10%CSE+1.0μmol·L-1番茄红素组和1.0μmol·L-1番茄红素组凋亡率分别为(6.2±0.5)%,(30.8±4.3)%,(18.3±1.9)%,(7.6±0.4)%。 qRT-PCR分析发现,与对照组比较,10%CSE组 SIRT1 mRNA表达是其(0.51±0.03)倍,10%CSE+1.0μmol·L-1番茄红素组为(0.84±0.05)倍、1.0μmol·L-1番茄红素组为(1.31±0.08)倍。 Western blot蛋白条带分析结果与qRT-PCR结果一致。结论番茄红素可以减轻CSE对HUVEC的损伤作用,其机制可能与番茄红素提高SIRT1表达有关。