[粒细胞集落刺激因子对高胆固醇喂养兔外周血内皮祖细胞的影响]

[Effects of granulocyte colony-stimulating factor on peripheral endothelial progenitor cells in cholesterol-fed rabbits].

作者信息

Zhao Wen-yan, Xiao Chuan-shi, Qiu Ling, Wang Gai-ling

机构信息

Department of Cardiology, Taiyuan People's Hospital, Taiyuan 030001, China.

出版信息

Zhonghua Xin Xue Guan Bing Za Zhi. 2006 Mar;34(3):257-61.

PMID:16630465

Abstract

OBJECTIVE

To investigate the effects of granulocyte colony-stimulating factor (G-CSF) on peripheral endothelial progenitor cells (EPC) and atherosclerosis (AS) in cholesterol-fed rabbits.

METHODS

Male New Zealand white rabbits were randomly divided into control group, G group (Recombinant Human Granulocyte Colony Stimulating Factor Injection rhG-CSF 50 microg/d), AS group (high cholesterol diet) and G + AS group (rhG-CSF 50 microg/d plus high cholesterol diet, n = 8 per group). Peripheral blood was collected at baseline and at 1, 4, 8 and 12 weeks, 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 being cultured for 7 days, EPCs were characterized as adherent cells double positive for DiLDL uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. After being cultured for 3 days, the number of EPC (PE-CD34/FITC-CD133 double-stained positive cells) was quantified by flow cytometric analysis. Serum NO was measured and aortic plaque area analyzed at 12 weeks.

RESULTS

EPC number was low in control and AS groups and EPC number was significantly increased ( approximately 13-fold, P < 0.001) compared to baseline at 1 week in G and G + AS groups and remained at this level throughout the study period in G group while decreased gradually in G + AS group and returned to baseline level at 12 weeks. Aortic atherosclerotic plaque was visible in both AS and G + AS groups, however, the aortic atherosclerotic plaque area was smaller in G + AS group than that of in As group (59.8 mm(2) +/- 26.9 mm(2) vs. 251.5 mm(2) +/- 83.4 mm(2), P < 0.01). Serum NO was similar between AS and G + AS groups and significantly higher than that in control and G groups.

CONCLUSION

CSF could attenuate atherosclerosis in cholesterol-fed rabbits by increasing circulating EPC.

摘要

目的

研究粒细胞集落刺激因子（G-CSF）对高脂喂养家兔外周血内皮祖细胞（EPC）及动脉粥样硬化（AS）的影响。

方法

雄性新西兰白兔随机分为对照组、G组（重组人粒细胞集落刺激因子注射液rhG-CSF 50μg/d）、AS组（高胆固醇饮食）和G+AS组（rhG-CSF 50μg/d加高胆固醇饮食，每组n=8）。于基线期及1、4、8和12周采集外周血，采用Ficoll密度梯度离心法从外周血中分离总单核细胞（MNC），然后将细胞接种于纤连蛋白包被的培养皿上。培养7天后，通过激光扫描共聚焦显微镜直接荧光染色，将EPC鉴定为对DiLDL摄取和凝集素结合呈双阳性的贴壁细胞。培养3天后，通过流式细胞术分析定量EPC数量（PE-CD34/FITC-CD133双染阳性细胞）。于12周时检测血清NO并分析主动脉斑块面积。

结果

对照组和AS组EPC数量较低，G组和G+AS组在第1周时EPC数量较基线显著增加（约13倍，P<0.001），G组在整个研究期间维持该水平，而G+AS组逐渐下降，在12周时恢复至基线水平。AS组和G+AS组均可见主动脉粥样硬化斑块，然而，G+AS组主动脉粥样硬化斑块面积小于AS组（59.8mm²±26.9mm² vs. 251.5mm²±83.4mm²，P<0.01）。AS组和G+AS组血清NO相似，且显著高于对照组和G组。