Zhu Jun-Hui, Tao Qian-Min, Chen Jun-Zhu, Wang Xing-Xiang, Zhu Jian-Hua, Shang Yun-Peng
Department of Cardiovasology, the First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou 310003, China.
Sheng Li Xue Bao. 2004 Jun 25;56(3):357-64.
The aim of the present study was to investigate whether fluvastatin augments the number of endothelial progenitor cells (EPCs), and promotes EPCs proliferation, migration and adhesion. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation. The cells were then plated on fibronectin-coated culture dishes. After being cultured for 7 d, the attached cells were stimulated with fluvastatin (final concentrations: 0.01, 0.1, 1, 10 micromol/L), simvastatin (1 micromol/L) or a vehicle for the respective time points (6, 12, 24 and 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs were further documented by demonstrating the expression of KDR, VEGFR-2 and AC133 with flow cytometry. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed by MTT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating it on fibronectin-coated dishes, and the adherent cells were then counted. In addition, we also studied EPCs culture assay of peripheral blood from fluvastatin-treated animals in vivo. Incubation of isolated human MNCs with fluvastatin dose- and time-dependently increased the number of EPCs, while reached the maximum 24 h after the administration at 1 micromol/L, (2.5-fold increase, P<0.05). Moreover, treatment of rats with fluvastatins elevated the number of EPCs (3-fold increase, P<0.05), thus extending the in vitro data. In addition, fluvastatin also promoted EPC proliferation, migration, adhesion and in vitro vasculogenesis in a concentration-dependent manner. The effects of fluvastatin on EPCs were compared with those of simvastatin at the same concentration (1 micromol/L), with a result of no statistical difference. The results of the present study define a novel mechanism of the action of statins: the augmentation of EPCs with enhanced functional activity.
本研究的目的是探讨氟伐他汀是否能增加内皮祖细胞（EPCs）的数量，并促进EPCs的增殖、迁移和黏附。通过Ficoll密度梯度离心从外周血中分离出总单核细胞（MNCs）。然后将细胞接种在纤连蛋白包被的培养皿上。培养7天后，用氟伐他汀（终浓度：0.01、0.1、1、10 μmol/L）、辛伐他汀（1 μmol/L）或相应时间点的溶剂对贴壁细胞进行刺激（6、12、24和48小时）。通过激光扫描共聚焦显微镜下的直接荧光染色，将EPCs鉴定为对DiLDL摄取和凝集素结合呈双阳性的贴壁细胞。通过流式细胞术检测KDR、VEGFR - 2和AC133的表达，进一步对EPCs进行记录。分别通过MTT法、改良Boyden小室法和体外血管生成试剂盒检测EPCs的增殖、迁移和体外血管生成活性。通过将其重新接种在纤连蛋白包被的培养皿上进行EPCs黏附试验，然后对贴壁细胞进行计数。此外，我们还研究了体内氟伐他汀治疗动物外周血的EPCs培养试验。用氟伐他汀孵育分离的人MNCs，EPCs数量呈剂量和时间依赖性增加，在1 μmol/L给药后24小时达到最大值（增加2.5倍，P<0.05）。此外，用氟伐他汀治疗大鼠可增加EPCs数量（增加3倍，P<0.05），从而扩展了体外数据。此外，氟伐他汀还以浓度依赖性方式促进EPCs的增殖、迁移、黏附和体外血管生成。将氟伐他汀对EPCs的作用与相同浓度（1 μmol/L）的辛伐他汀进行比较，结果无统计学差异。本研究结果确定了他汀类药物作用的一种新机制：增强功能活性的EPCs数量增加。
