粒细胞集落刺激因子与动脉粥样硬化对外周血内皮祖细胞数量的影响  

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

作　　者：赵文燕[1] 肖传实[1] 邱龄[1] 王改玲[1] 

机构地区：[1]山西医科大学第二医院心内科

出　　处：《中华心血管病杂志》2006年第3期257-261,共5页Chinese Journal of Cardiology

基　　金：吴阶平医学基金会临床科研资助项目(2003-97-A)

摘　　要：目的观察粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)与动脉粥样硬化(atherosclerosis,AS)对外周血内皮祖细胞(EPC)的影响。方法将32只雄性新西兰白兔随机分为G组(重组人粒细胞集落刺激因子rhG-CSF50μg/d)、G+AS组(rhG-CSF50μg/d、高脂饲料)、AS组(高脂饲料)及对照组,每组各8只。4组动物分别于实验前及第1、4、8、12周采血,培养7天后,用荧光显微镜观察鉴定FITC-UEA-I和Dil-acLDL双染色阳性细胞为正在分化的EPC;细胞培养3天后,通过流式细胞仪计数各组PE-CD34、FITC-CD133双阳性细胞为EPC;第12周测血清一氧化氮、血脂水平并做主动脉斑块分析。结果实验前,各组EPC含量均很低;用G-CSF治疗1周,G组及G+AS组EPC迅速升高(与用药前比较增加了约13倍,P<0·001),继续给药G组EPC维持在一个较高水平(第1、4、8、12周比较P>0·05);给予高脂饮食后,G+AS组EPC数量逐渐下降(第4、8周与对照组比较P<0·001、P<0·001;第12周与对照组比较P=0·326);对照组EPC一直处于低水平,AS组EPC数量较对照组低,但两组各周比较P>0·05。经过12周的高脂喂养,G+AS组及AS组主动脉均形成了动脉粥样硬化斑块,但G+AS组斑块面积、斑块/内膜面积均低于AS组(P<0·01)。第12周G+AS组及AS组一氧化氮含量均低于G组和对照组(P<0·001)。结论EPC与动脉粥样硬化关系密切,AS损害内皮,减少EPC,G-CSF对EPC有动员作用,能够增加外周血EPC数量,因而对血管有保护作用,抑制AS进展。Objective To investigate the effects of granulocytecolony-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 p.g/d）, AS group （high cholesterol diet） and G＋AS group （rhG-CSF 50 p.g/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 FieoU density gradient centrifugation, and then the cells were plated on fibronectincoated 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 confecal microscope. After being cultured for 3 days, the number of EPC （PE -CD34/ FITC-CD133 double-stained positive cells） was quantified by flow eytometrie 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 （ - 13-fold, P 〈 O. 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 mm2, 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 atherosclemsis in cholesterol-fed rabbits by increasing circulating EPC.

关 键 词：动脉硬化 粒细胞集落刺激因子 干细胞 

分 类 号：R543.5[医药卫生—心血管疾病]