内皮祖细胞动员及与冠心病危险因素关系的实验性研究

【作者】 赵文燕；

【导师】 肖传实； 邱龄；

【作者基本信息】 山西医科大学 ， 心血管内科学， 2005， 硕士

【摘要】 第一章 非诺贝特对兔外周血内皮祖细胞数量的影响 目的 以阿托伐他汀为对照观察不同剂量非诺贝特对外周血内皮祖细胞(endothelial progenitor cell, EPC)数量的影响,进一步探讨非诺贝特对 EPC 是否有动员作用。 方法 将 32 只雄性新西兰白兔随机分为非诺贝特 1 组(50 ㎎/㎏/d)、非诺贝特 2 组(25 ㎎/㎏/d)、阿托伐他汀组(5 ㎎/㎏/d)及对照组,每组各 8 只。4 组动物分别于实验前及第 1、2、3、4 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。 结果 实验前,各组 EPC 含量均很低;用药 1 周,阿托伐他汀组 EPC 增加了约 7 倍(P<0.01),第 3 周增加到 20 倍(P<0.001);非诺贝特 1 组、2 组在整个 4 周的观察期间 EPC 无明显增加(P>0.05)。结论 非诺贝特(50 ㎎/㎏/d、25 ㎎/㎏/d)对 EPC无动员作用。 第二章 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 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-CSF 50μg /d)、G+As 组(rhG-CSF 50μg /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 双山西医科大学硕士学位论文 II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 12W 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、12W 比较 P>0.05);给予高脂饮食后,G+As 组 EPC 数量逐渐下降(第 4、8W 与对照组比较 P<0.001、P<0.001;第 12W 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 12W 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)。G+As 组与 As 组血脂各项指标均明显高于对照组(P<0.001),两组间比较(P>0.05)。第 12W G+As 组及 As 组 NO 含量均低于 G 组和对照组(P<0.001),二组比较(P<0.05) 。 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。关键词: 粒细胞集落刺激因子;动脉粥样硬化;内皮祖细胞第三章 粒细胞集落刺激因子与糖尿病对外周血内皮祖细胞数量的影响目的 观察粒细胞集落刺激因子(granulocyte colony-stimulatingfactor, G-CSF) 与 糖 尿 病 (DM) 对 外 周 血 内 皮 祖 细 胞 (endothelialprogenitor cell, 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-CSF 50μg/d)、G+DM 组(糖尿病模型、rhG-CSF 50μg/d)、DM 组(糖尿病模型)及对照组,每组各 8 只。4 组动物分别于实验前及第 1、2、3、4 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;细胞培养更多还原

【Abstract】 PartⅠ Effects of fenofibrate on the number of endothelial progenitor cells from peripheral blood in rabbit Objective To investigate whether fenofibrate have influence on endothelial progenitor cells (EPC) from peripheral blood contrast to atorvastatin group. Methods 32 male New Zealand white rabbits were divided into fenofibrate group one(50 ㎎/㎏/d), fenofibrate group two(25 ㎎/㎏/d)and control group, there were 8 rabbits in each groups. Before treatment and 1.2.3.4w, peripheral blood was collected. After 7 days cultured, EPC were characterized as double positive for DiLDL-uptake and lectin binding by direct fluorescent staining. After 3 days cultured, the number of EPC (PE -CD34/ FITC-CD133 dual-stained positive cells) was surveyed by Flow Cytometry Analysis . Results Before treatment, the number of EPC was low; After a week treatment of atorvastatin, the number of EPC in atorvastatin was higher than before (≈7-fold, P<0.01), and kept significant higher in 3w (≈20-fold, P<0.001). The number of EPC in fenofibrate group one and two was low throughout the 4-week study period P>0.05. Conclusion fenofibrate (50 ㎎/㎏/d, 25 ㎎/㎏/d) could not mobilize EPC . PartⅡ Effects of atherosclerosis and granulocyte colony-stimulating factor on the number of endothelial progenitor cells from peripheral blood Objective To investigate whether atherosclerosis (As) and granulocyte colony-stimulating factor (G-CSF) have influence on endothelial progenitor cells (EPC) from peripheral blood. Methods 32 male New Zealand white rabbits were divided into G group(Recombinant Human Granulocyte Colony Stimulating Factor Injection rhG-CSF 50μg /d), G+As group (rhG-CSF 50μg/d, high cholesterol diet), As group (high cholesterol diet) and control group, there were 8 rabbits in each groups. Before treatment and 1.4.8.12w, peripheral blood was collected. After 7 days cultured, EPC were characterized as double positive for DiLDL-uptake and lectin binding by direct fluorescent staining. After 3 days cultured, the number of EPC (PE -CD34/ FITC-CD133 dual-stained positive cells) was surveyed by Flow Cytometry Analysis . At 12th week,serum NO and lipids were measured and square of the aortic plaque were researched. Results Before treatment, the number of EPC was low; After a week treatment of G-CSF, the number of EPC in G group and G+As group was fast higher than before (≈13-fold, P<0.001), and kept persistence significant higher in G group in 12w (1.4.8.12W compared each other P>0.05);After fed on high cholesterol diet, the number of EPC in G+As group decreased gradually. At 4th and 8th week, compared with control group, respectively P<0.001, P<0.001;At 12th week, compared with control group P=0.326. The number of EPC in control group had kept low in the whole process. The number of EPC in As group was lower than control group, but P>0.05. After fed on high cholesterol diet, there were aortic plaque in G+As group and As group, but the square of the aortic plaque was significant difference, P<0.01. Conclusion As could impair endothelial cells and reduce the number of EPC. G-CSF could mobilize EPC, improve the number of EPC in peripheral blood, protect blood vessel and prevent As. Part Ⅲ Effects of diabetes mellitus and granulocyte colony-stimulating factor on the number of endothelial progenitor cells from peripheral blood Objective To investigate whether diabetes mellitus (DM) and granulocyte colony-stimulating factor (G-CSF) have influence on endothelial progenitor cells (EPC) from peripheral blood. Methods 32 male New Zealand white rabbits were divided into G group(rhG-CSF 50μg /d), G+DM group (rhG-CSF 50μg/d, DM model), DM group(DM model)and control group, there were 8 rabbits in each groups. Before treatment and 1.2.3.4w, peripheral blood was collected. After 7 days cultured, EPC were characterized as double positive for DiLDL-uptake and lectin binding by direct fluorescent staining. After 3 days cultured, the number of EPC (PE-CD34/ FITC-CD133 dual-stained positive cells) was surveyed by Flow Cytometry Analysis. At 4th week, seru更多还原