氨氯地平对ox-LDL致大鼠骨髓EPCs损伤的保护作用与上调一氧化氮合酶表达有关

Antagonism Effect of Amlodipine on Oxidative Low Density Lipoprotein Injury Rat Marrow Endothelial Progenitor Cells is Related to Up-regulation of e-NOS Expression

目的探讨氨氯地平对氧化型低密度脂蛋白(ox-LDL)致大鼠骨髓内皮祖细胞(EPCs)损伤的保护作用。方法实验分为对照组、ox-LDL组、氨氯地平组(0.1、0.5、1.0μmol/L),贴壁法培养分离内皮祖细胞,细胞免疫荧光鉴定EPCs,MTT法检测细胞活性,Western blotting检测一氧化氮合酶(eNOS)蛋白表达水平。结果贴壁法能分离纯度达70%以上的CD133+/VEGFR-2+双阳性EPCs;0.5μmol/L氨氯地平可显著拮抗ox-LDL对EPCs增殖的抑制作用;0.5μmol/L氨氯地平能显著上调eNOS蛋白表达(P<0.01)。结论氨氯地平能拮抗ox-LDL对EPCs的损伤,氨氯地平的拮抗作用与上调eNOS表达有关。

Objective To investigate the protective effect of amlodipine on oxidative low density lipoprotein treated rat marrow endothelial progenitor cells（EPCs）.Methods EPCs was isolated from rat marrow,and was divided into three groups,control,ox-LDL group and Amlodipine group.Adhesion method was applied to isolate EPCs,immunofluorescence was used to identify EPCs,and MTT for proliferation,western blotting were used to measure protein of eNOS.Results Almost 70 percent of CD133＋/VEGFR-2＋ double positive EPCs can be isolated from rat marrow by adhesion method;Antagonism effect of amlodipine on proliferation of EPCs was found at the concentration 0.5 μmol/L,and this inhibitory effect can be recovered by 0.5 μmol/L amlodipine;eNOS was reduced by 50 μg/mL ox-LDL,and it can be restored remarkably by administrating 0.5 μmol/L amlodipine.Conclusions Injury effect of ox-LDL can be antagonism by amlodipine,this protective effect of amlodipine is related to up-regulating eNOS expression.