中国中药杂志

2004, (08)

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葛根素对外周血内皮祖细胞数量和功能的影响
Effects of puerarin on number and activity of endothelial progenitor cells from peripheral blood

张芙荣,陈君柱,朱军慧,王兴祥,朱建华,尚云鹏,郭晓纲,戴禾敏,孙坚
ZHANG Fu-rong, CHEN Jun-zhu, ZHU Jun-hui, WANG Xing-xiang, ZHU Jian-hua, SHANG Yun-peng, GUO Xiao-gang, DAI He-min, SUN Jian(Department of Cardiovascular Diseases, First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou 310003, China)

摘要(Abstract):

目的 :观察葛根素对外周血内皮祖细胞的影响。方法 :采用密度梯度离心法从外周血获取单个核细胞 ,将其接种在人纤维连接蛋白包被培养板 ,培养 7d后 ,收集贴壁细胞 ,加入不同浓度葛根素 (分别为 0 .1,0 .5 ,1,3mmol·L-1)培养一定的时间 (6 ,12 ,2 4 ,4 8h)。通过激光共聚焦显微镜鉴定FITC UEA Ⅰ和DiI acLDL双染色阳性细胞被认为是正在分化的EPC ,并在倒置荧光显微镜下计数。然后分别采用MTT比色法、改良的Boyden小室、黏附能力测定实验和体外血管生成试剂盒观察EPC的增殖能力、迁移能力、黏附能力和体外血管生成能力。结果 :葛根素显著增加外周血EPC数量 ,并且EPC数量随葛根素浓度增加及作用时间延长而增加 ,3mmol·L-1浓度葛根素作用 2 4h对EPC数量的影响最为显著 (较对照组增加了 1倍 ,P <0 .0 1)。葛根素也显著改善了外周血EPC的黏附能力、迁移能力、增殖能力和体外血管生成能力。结论 :结果提示葛根素可增加EPC的数量且伴随着EPC功能的改善
Objective: To investigate whether puerarin can augment endothelial progenitor cells (EPCs) numbers, promote EPC proliferation, migration and adhesion. Method: 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 culture, attached cells were stimulated with puerarin (to make a series of final concentrations: 0.1, 0.5, 1, 3 mmol·L~(-1)) or vehicle control for the respective time points(6, 12, 24, 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 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 those on fibronectin-coated dishes, then adherent cells were counted. Result: Incubation of isolated human MNCs with puerarin dose increased the number of EPCs, maximum at 3 mmol·L~(-1), 24 hours (approximately 1-fold increase, P<0.01). In addition, puerarin also promoted EPC proliferative, migratory,adhesive and in vitro vasculogenesis capacity. Conclusion: Puerarin can augmentf the number of EPCs with enhanced functional activity.

关键词(KeyWords): 冠心病;内皮祖细胞;葛根素;激光共聚焦显微镜
coronary artery disease; endothelial progenitor cell; puerarin; laser scanning confocal microscope

Abstract:

Keywords:

基金项目(Foundation): 浙江省科学技术研究基金项目 ( 0 2 110 7613 ) ;; 浙江省卫生科学研究基金项目 ( 2 0 0 3A0 43 )

作者(Author): 张芙荣,陈君柱,朱军慧,王兴祥,朱建华,尚云鹏,郭晓纲,戴禾敏,孙坚
ZHANG Fu-rong, CHEN Jun-zhu, ZHU Jun-hui, WANG Xing-xiang, ZHU Jian-hua, SHANG Yun-peng, GUO Xiao-gang, DAI He-min, SUN Jian(Department of Cardiovascular Diseases, First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou 310003, China)

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