芹菜素延缓人脐静脉内皮细胞衰老的体外实验研究

Apigenin Delays the Senescence of Human Umbilical Vein Endothelial Cells in Vitro

目的:探究芹菜素对人脐静脉内皮细胞衰老的延缓作用及相关机制。方法:将人脐静脉内皮细胞培养至第12代(老年细胞),且自第4代(幼年细胞)起,与芹菜素(1、3、10μmol·L-1)共孵育。通过检测细胞β-半乳糖苷酶(SAβ-gal)阳性率,推断细胞衰老程度;通过检测胞内双氢罗丹明氧化产物Rh123的荧光强度,推断胞内活性氧簇(ROS)水平,即氧化应激水平;通过检测培养液中硝酸盐/亚硝酸盐含量,推断胞内一氧化氮(NO)水平。结果:与幼年细胞组相比,老年细胞对照组中内皮细胞SAβ-gal阳性率及ROS水平均明显提高(P<0.05),且其培养液中硝酸盐/亚硝酸盐含量明显降低(P<0.05);在老年细胞组中,与对照组相比,芹菜素组中内皮细胞SAβ-gal阳性率及ROS水平均明显呈芹菜素剂量依赖性降低(P<0.05),而其培养液中硝酸盐/亚硝酸盐含量也明显呈芹菜素剂量依赖性增加(P<0.05)。结论:芹菜素可延缓人脐静脉内皮细胞的衰老,且该作用与胞内ROS生成减少及NO生成增加有关。

Objective： To explore the effect of apigenin on delaying the senescence of human umbilical vein endothelial ceils and the related mechanism. Methods： Human umbilical vein endothelial cells were cultured until the twelfth passage （old cells） and co-incubated with apigenin （1,3, 10 μmol L^-1） starting at the fourth passage （young cells）. The cell senescence was measured by the determination of the fl-galactosidase （SA β-gal） positive rate of the cells; the intracelluar ROS level （oxidative stress） was measured by the determination of the fluorescent intensity of Rh123-an oxidative product of dihydrorhodamin in the cells; the intracelluar NO level was measured by the determination of the content of nitrate/ nitrite in the cell culture. Results： The SAβ-gal positive rate of the cells and intraeellular ROS level in old cell control group were significantly increased and the content of nitrate/nitrite in the cell culture was significantly decreased, compared with those in young cells group （P 〈 0. 05）. In old cells group, apigenin dose-dependently decreased the SA fl-gal positive rate of the cells and intracellular ROS level and increased the content of nitrate and nitrite in the cell culture, compared with the untreated control cells （P 〈0. 05）. Conclusion： Apigenin could delay the senescence of human umbilical vein endothelial cells, and this effect was related to the decrease of intracellular ROS generation and the increase of NO synthesis in the cells.