丹参素对氧化低密度脂蛋白损伤内皮祖细胞的影响及机制探讨

Effects of Danshensu on function of EPCs which were damaged by Ox-LDL and study its possible mechanism

目的:观察丹参素对氧化低密度脂蛋白(oxidative low density lipoprotein Ox-LDL)损伤后外周血内皮祖细胞(en-dothelial progenitor cells,EPCs)功能的影响,并探讨其可能的机制。方法:密度梯度离心法获取外周血单个核细胞,通过流式细胞仪鉴定内皮祖细胞。培养7 d后,收集贴壁细胞并随机分为对照组,氧化低密度脂蛋白组(100 mg.L-1)及丹参素干预组(氧化低密度脂蛋白100 mg.L-1加丹参素,浓度分为2,10,50 mg.L-1),干预24 h后分别采用MTT比色法、黏附能力测定实验观察EPC的增殖能力、黏附能力。并取各组细胞上清液行超氧化物歧化酶(SOD)、丙二醛(MDA)含量检测。结果:氧化低密度脂蛋白损伤后,外周血EPCs的增殖能力、黏附能力显著受损,细胞上清液SOD含量显著下降,MDA含量显著升高;丹参素干预24 h后,显著改善了外周血EPCs的功能,各组SOD含量显著增加,MDA含量显著减少。结论:丹参素对氧化低密度脂蛋白损伤后内皮祖细胞的功能有显著保护作用,其机制可能与抗氧化损伤有关。

Objective： To observe the effects of Danshensu on function of endothehal progenitor cells （EPCs） from peripheral blood which were damaged by oxidative low density lipoprotein （Ox-LDL）. And study its possible mechanism. Method： Total mononuclear cells （ MNCs ） were isolated from peripheral blood by ficoll density gradient centrifugation, and were identified by demonstrating the expression of CD34, VEGFR-2 and AC133 with flow cytometry, to sure that all the cells needed were EPCs. Then the cells were plated on fibronectin-coated culture dishes. After incubation for 7 days, attached cells were collected and divided into three groups： Control group, Ox-LDL group, Danshensu intervention group, stimulated with different cencentrations of Danshensu （2, 10 and 50 mg · L-1 ）, adhesion assay respectively. EPCs adhesion assay were performed by replating those on fibronectin-coated dishes, then adherent cells were counted. And take cell supemate of each group to carry on the SOD, MDA content examination. Result： Ox-LDL impaired EPC prohferative and adhesive capacity. In Ox-LDL group, The SOD content obviously drops, the MDA content obviously elevates. After Danshensu interventing for 24 h, adhesive EPCs and migratory EPCs were significantly increased, compared with Ox-LDL group, The SOD content of Danshensu intervention group obviously increased and the MDA content obviously reduced. Conclusion： Danshensu could improve prohferative and adhesive capacity of EPCs that were impaired by Ox-LDL. The mechanism might relate to the oxidation resistance damage.