Wang Xing-xiang, Shang Yun-peng, Chen Jun-zhu, Zhu Jun-hui, Guo Xiao-gang, Sun Jian
Department of Cardiovascular Diseases, the First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou 310003, China.
Yao Xue Xue Bao. 2004 Aug;39(8):656-60.
To investigate whether Ginkgo biloba extract can augment endothelial progenitor cell (EPC) number, and promote EPC proliferation, migration and adhesion.
Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days of culture, attached cells were stimulated with Ginkgo biloba extract (10, 25 and 50 mg x L(-1)) or vehicle control for the respective time points (6, 12, 24 and 48 h). EPC were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPC were further documented by demonstrating the expression of CD34, VEGFR-2 and AC133 with flow cytometry. EPC proliferation, migration and in vitro vasculogenesis activity were assayed with MTT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating MNCs on fibronectin-coated dishes, and then counting adherent cells.
Incubation of isolated human MNCs with Ginkgo biloba extract increased the number of EPC, maximum at 25 mg x L(-1), 24 hours (approximately 1-fold increase, P < 0.01). In addition, Ginkgo biloba extract promotes EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity.
Ginkgo biloba may promote EPC augmentation and enhance its functional activity.
研究银杏叶提取物是否能增加内皮祖细胞(EPC)数量,并促进EPC增殖、迁移和黏附。
采用Ficoll密度梯度离心法从外周血中分离出总单核细胞(MNCs),然后将细胞接种于纤连蛋白包被的培养皿上。培养7天后,对贴壁细胞分别用银杏叶提取物(10、25和50 mg·L⁻¹)或溶剂对照刺激相应时间点(6、12、24和48小时)。在激光扫描共聚焦显微镜下通过直接荧光染色将EPC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CD34、VEGFR-2和AC133的表达进一步对EPC进行鉴定。分别用MTT法、改良Boyden小室法和体外血管生成试剂盒检测EPC的增殖、迁移和体外血管生成活性。通过将MNCs重新接种到纤连蛋白包被的培养皿上,然后计数贴壁细胞来进行EPC黏附试验。
用银杏叶提取物孵育分离的人MNCs可增加EPC数量,在25 mg·L⁻¹、24小时时达到最大值(增加约1倍,P<0.01)。此外,银杏叶提取物可促进EPC的增殖、迁移、黏附及体外血管生成能力。
银杏叶可能促进EPC增加并增强其功能活性。