冠心病患者循环内皮祖细胞与血管内皮功能的变化

目的:探讨冠心病患者循环内皮祖细胞(EPCs)数量及功能变化与血管内皮舒张功能的关系。方法:将58例患者分为对照组(20例)、稳定性心绞痛组(11例)、不稳定性心绞痛组(27例)。采用高分辨率二维超声检测肱动脉血流介导的内皮依赖性血管舒张功能(FMD)及硝酸甘油介导的非内皮依赖性血管舒张功能(NMD);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7天后贴壁细胞进行细胞化学分析,激光共聚焦显微镜鉴定异硫氰酸荧光素标记荆豆凝集素I和DiI标记的乙酰化低密度脂蛋白双染色阳性细胞为正在分化的EPCs,采用二苯基四氮唑嗅盐比色法、改良的Boyden小室和黏附能力测定实验观察内皮祖细胞的增殖能力、迁移能力和黏附能力。结果:①稳定性心绞痛组与不稳定性心绞痛组的FMD均明显低于对照组,有显著性差异(P<0．05～0．01),而稳定性心绞痛组的FMD较不稳定性心绞痛组也降低,有显著性差异(P<0．05);3组间NMD差异无统计学意义(P>0．05)。②稳定性心绞痛组、不稳定性心绞痛组较对照组循环EPCs数量明显减少,且黏附、迁移及增殖能力也明显下降,均有极显著性差异(P<0．01);不稳定性心绞痛组较稳定性心绞痛组EPCs数量及迁移能力无差异(P>0．05),但黏附和增殖能力降低(P<0．05)。③直线相关性分析发现不稳定性心绞痛组EPCs的数量及功能均与FMD呈正相关(P<0．05),而稳定性心绞痛组仅EPCs黏附功能与FMD呈正相关(P<0．05)。结论:冠状动脉EPCs数量及功能下降与血管内皮依赖性舒张功能障碍一致,提示当冠心病患者血管内皮功能受损而又缺乏足够有效的EPCs时,可能影响冠心病的病情程度及临床表现。     

高血压亚急症患者血管功能异常及与循环内皮祖细胞功能变化的关系

目的探讨高血压亚急症患者循环内皮祖细胞(EPC)功能的改变以及一氧化氮(NO)、肱动脉血流介导的内皮依赖性舒张功能(FMD)与EPC功能的关系。方法招募高血压亚急症患者13例,血压正常者20例(对照组),取外周血提取原代EPC培养后以Transwell小室和CCK-8法评估EPC的迁移、黏附和增殖能力,通过检测FMD、血浆和EPC分泌的NO、粒-巨噬细胞集落刺激因子(GM-CSF)、血管内皮生长因子(VEGF)、白细胞介素6(IL-6)水平评估血管内皮功能。结果高血压亚急症组与对照组相比,EPC细胞功能(包括迁移、增殖和黏附能力)明显下降(P0.05);血浆及EPC分泌的NO水平有明显差异(P0.05);血浆及EPC分泌NO水平均与EPC迁移、黏附和增殖能力呈明显的线性关系(P0.05);FMD与EPC迁移、黏附和增殖能力具有明显相关性(P0.05)。两组血浆和EPC分泌的GM-CSF、VEGF、IL-6水平均无明显差异(P0.05)。结论相对血压正常者,高血压亚急症人群的EPC功能(迁移、黏附和增殖)明显下降,且NO水平和FMD均与EPC功能正相关。     

增龄对内皮祖细胞数量和功能活性的影响及与血管内皮功能的关系

目的探讨增龄对人外周血内皮祖细胞数量和功能活性的影响及与血管内皮功能的关系。方法入选32例健康老年人作为实验组(老年组)和30例健康青年人为对照组(青年组)。抽取受试者外周静脉血20 ml,采用密度梯度离心法分离提取内皮祖细胞,采用流式细胞技术比较两组志愿者循环中内皮祖细胞数量。体外培养7 d后,分别检测两组志愿者内皮祖细胞体外迁移及黏附能力;通过裸鼠颈动脉拉脱模型,比较内皮祖细胞在体内皮损伤修复能力。检测两组志愿者血流介导的血管舒张功能(FMD)。结果与青年组相比,老年组外周血中内皮祖细胞数量明显下降(P0.01),体外迁移、黏附(P0.01)及在体再内皮化功能均明显受损。减少的在体再内皮化面积与下降的FMD之间呈正相关。结论增龄导致外周血中内皮祖细胞数量和功能活性受损与血管内皮功能下降密切相关,是动脉粥样硬化血管疾病发生发展的重要环节。     

冠心病患者外周血内皮祖细胞数量和功能的变化

目的 :观察冠心病 (CHD)患者外周血内皮祖细胞 (endothelialprogenitorcells,EPC)数量和功能的改变。方法 :选择CHD患者和非CHD患者各 2 0例 ,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后贴壁细胞进行细胞化学分析。激光共聚焦显微镜鉴定FITC标记荆豆凝血素Ⅰ和DiI 标记的乙酰化低密度脂蛋白双染色阳性细胞为正在分化的EPC。采用二苯基四氮唑嗅盐比色法、改良的Boyden小室和黏附能力测定实验观察EPC的增殖能力、迁移能力和黏附能力。 结果 :CHD患者外周血EPC数量明显减少 [(31.8± 7.7)∶(5 9.5± 10 .6 )EPC/视野 (× 2 0 0 ) ,P <0 .0 5 ],且CHD患者外周血EPC的黏附能力、迁移能力和增殖能力也明显受损。结论 :CHD患者外周血EPC数量减少、功能减退。     

冠心病患者内皮祖细胞数量及功能变化的研究进展

冠心病(CHD)是现代社会严重威胁人类健康的主要疾病之一,且病死率高。动脉粥样硬化（AS）是其病理基础。而内皮细胞的损伤与CHD的发生发展关系密切。血管内皮祖细胞(endothelial progenitor cells,EPCs)作为内皮细胞的前体细胞,在组织缺血及血管损伤时动员入血,参与微血管的生成及血管内皮的修复,在冠心病发生及发展过程中起着非常重要的作用。已经证明EPCs数量下降及功能减退与CHD发病有密切关系,现将冠心病患者外周血内皮祖细胞数量及功能的变化情况做一详细阐述。     

糖尿病时循环内皮细胞、内皮祖细胞和循环祖细胞的变化及其临床意义

糖尿病（DM）患者，特别在并发大血管病和微血管病时，体内循环内皮细胞（CECs）、内皮祖细胞（EPCs）和循环祖细胞（CPCs）均发生数量和功能的变化。本文将围绕这些变化讨论其发生原因、可能机制及临床意义。     

高胆固醇对内皮祖细胞数量和功能的影响

目的　观察高胆固醇血症患者外周血内皮祖细胞 (EPC)数量和功能的变化。方法选择高胆固醇血症患者和非高胆固醇血症患者各 2 0例 ,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后贴壁细胞进行细胞化学分析。激光共聚焦显微镜鉴定FITC UEA Ⅰ和DiI acLDL双染色阳性细胞为正在分化的EPC。采用MTT比色法、改良的Boyden小室和黏附能力测定实验观察EPC的增殖、迁移和黏附能力。结果　高胆固醇血症患者外周血EPC数量明显减少 [(41 8± 8 7比 6 4 5± 16 6 )EPCs/× 2 0 0视野 ,P <0 0 5 ],且EPC数量与血总胆固醇水平 (r =- 0 6 5 9,P <0 0 0 1)和低密度脂蛋白胆固醇水平 (r =- 0 6 11,P <0 0 0 1)呈反向线性关系。高胆固醇血症患者EPC的增殖、迁移和黏附能力也明显受损。结论　高胆固醇患者外周血内皮祖细胞数量减少、功能减退。     

内皮祖细胞的功能及应用研究进展

内皮祖细胞(endothelial progenitor cells,EPCs),主要生成和存在于骨髓中,是一类干细胞和血管内皮细胞(ECs)之间具有游走特性的能够自我更新和增殖分化的定向干细胞.未分化的EPCs呈圆形,已分化的EPCs呈梭形或纺锤形.未分化的EPCs在形态上很难与其它细胞区分开来,目前主要依靠细胞表面标志来区别,多数学者认为同时具有CD34+、AC133+及VEGFR-2(血管内皮生长因子受体2)等表面抗原的称为EPCs[1].新近研究表明,起源于脐血单核细胞的CD34+/CD14-,或CD34-/CD14+的细胞,也可分化为ECs[2].近年来的研究表明eNOS(内皮型一氧化氮合酶),也是它的特征之一[3].上个世纪末,随着对EPCs诱导[4]、培养、鉴定、分化、迁移及归巢研究的广泛开展,有关EPCs应用的报道越来越多,呈现出从基础到临床蓬勃发展的形势,为血管外科治疗难治性血管病提供了新的理论基础和思路,本文就这一时期EPCs的功能及应用方面的研究进展作一总结.     

循环内皮祖细胞与心血管疾病

内皮祖细胞具有较强的增殖、分化潜能,能够分化为成熟内皮细胞等血管细胞,使其成为血管发生和血管新生领域研究的热点.自1997年,首次在循环血中发现内皮祖细胞以来,对循环内皮祖细胞(circulating endothelial progenitor cells,CEPCs)生物学特性有了进一步深入的认识和理解,它不仅是出生后血管形成的物质来源之一,而且还是保护和修复机体整个血管内皮层的重要力量.因此,CEPCs与冠心病、高血压等心血管疾病密切的联系同样受到研究者的重视.     

冠心病患者循环内皮祖细胞变化及功能的研究

目的探讨冠心病患者外周血循环内皮祖细胞(EPC)变化及其纤溶、黏附和炎症因子表达。方法选择冠心病患者57例(冠心病组)和对照组30例,提取EPC进行数量和细胞集落的比较。用酶联免疫吸附法和底物发光法检测EPC分泌组织型纤溶酶原激活物(tPA)和纤溶酶原激活物抑制剂(PAI)的浓度和活性。用反转录-聚合酶链反应(RT-PCR)检测EPC的tPA、PAI、血管细胞黏附分子1(VCAM-1)、细胞间黏附分子1(ICAM-1)、过氧化物酶体增殖因子活化受体γ(PPARγ)的mRNA表达水平。结果冠心病组EPC数量较对照组明显减少(P0.05),形成细胞集落数、细胞增殖能力也明显降低(P0.05)。与对照组比较,冠心病组EPC分泌的tPA含量和活性明显下降(P0.05),PAI含量和活性明显升高(P0.01)。RT-PCR检测结果显示,与对照组比较,冠心病组EPC的tPA、PPARγmRNA表达减弱,PAI、VCAM-1、ICAM-1 mRNA表达增强(P0.05)。结论冠心病患者外周血循环EPC数量减少,纤溶功能减低,黏附和炎症因子表达增强,其在冠心病发生发展中起到重要作用。     

Reduced number and impaired function of circulating endothelial progenitor cells in patients with abdominal aortic aneurysm

Aim

Circulating endothelial progenitor cells (EPCs) are associated with coronary artery disease (CAD) and predict its outcome. Although the pathophysiology of abdominal aortic aneurysm (AAA) is different, it shares some risk factors with CAD. Therefore, the correlation between EPCs and AAA was investigated.

Methods and results

Seventy-eight subjects (age 77.2 ± 7.8 years) with suspected AAA were prospectively enrolled. Cut-off values (men, 3.5–5.5 cm; women, 3–5 cm) were used to define normal aorta, small AAA, and large AAA on thoraco-abdominal computer tomography. Endothelial function was measured by flow-mediated vasodilation (FMD). Flow cytometry and colony-forming units (CFUs) were used to evaluate circulating EPC numbers. Circulating EPCs were defined as mononuclear cells with low CD45 staining and double-positive staining for KDR, CD34, or CD133. Late out-growth EPCs were cultured from six patients with large AAAs and six age- and sex-matched controls to evaluate proliferation, adhesion, migration, tube formation, and senescence.FMD was significantly lower with large (5.26% ± 3.11%) and small AAAs (6.31% ± 3.66%) than in controls (8.88% ± 4.83%, P = 0.008). Both CFUs (normal 38.39 ± 12.99, small AAA 21.22 ± 7.14, large AAA 6.98 ± 1.97; P = 0.026) and circulating EPCs (CD34⁺/KDR⁺ and CD133⁺/KDR⁺) were significantly fewer in AAA patients than in controls. On multivariate analysis, CFUs and circulating EPCs (CD34⁺/KDR⁺) were independently, inversely correlated to AAA diameter. Proliferation, adhesion, migration, tube formation, and senescence of late EPCs were significantly impaired in AAA patients.

Conclusion

The number and function of EPCs were impaired in AAA patients, suggesting their potential role in AAA.     

The association between circulating endothelial progenitor cells and coronary collateral formation

Aim

We investigated the relationship between coronary collateral formation and circulating endothelial progenitor cells (EPC) in patients undergoing coronary angiography.

Methods and results

Circulating CD133⁺/34⁺ and CD34⁺/KDR⁺ EPCs were determined in 68 patients (normal coronary vessels in 24 patients and coronary artery disease (CAD) in 44 patients) (age: 58.7 ± 10.1, 64.7% male). Circulating EPCs were higher among patients with normal coronary vessels compared to patients with CAD for CD133⁺/34⁺ (p < 0.05) and CD34⁺/KDR⁺ cells (p < 0.05). The number of EPCs were significantly greater in patients with good coronary collateral formation (p < 0.05). EPC count was independent predictor for coronary collateral formation after adjustment for other cardiovascular risk factors and extent of CAD (p = 0.037).

Conclusion

In patients with severe coronary stenosis, those with increased circulating EPCs had better collateral formation compared to those with lower EPC counts. Our findings implicate that in addition to presence of critical stenosis, intact response of bone marrow is necessary for collateral formation in CAD.     

活动期系统性红斑狼疮患者外周血内皮祖细胞数量及功能变化的研究

目的 观察活动期系统性红斑狼疮(SLE)患者外周血内皮祖细胞(CEPC)的数量、功能(包括抗炎功能)以及整体状态的变化.方法 研究共纳入35例病情处于活动期的SLE患者(女性28例,男性7例),另选择35名年龄和性别与之匹配的健康志愿者作为对照组.CEPC的数量通过荧光激活细胞分选方法确定;CEPC在纤维连接蛋白上的黏附能力以及单核细胞系细胞(THP-1细胞)向CEPC的黏附能力均采用细胞黏附计数法进行评价;CEPC的迁移能力通过Transwell小室迁移率分析法来分析;采用离体Matrigel上成管实验方法检测CEPC的成管功能.分别采用实时定量聚合酶链反应(PCR)法和Western-blotting法测定CEPC中诱导型一氧化氮合酶(iNOS)和白细胞介素(IL)-6的表达,以及细胞黏附分子(ICAM)-1和磷酸化akt(p-akt)的表达(后者反映CEPC的抗炎能力和整体状态).结果 活动期SLE患者CEPC的数量与对照组比较差异无统计学意义[(0.054±0.018)%和(0.079±0.017)%.P=0.094],但其部分功能受影响:CEPC在纤维连接蛋白上的黏附能力显著下降(50±17和24±11,P=0.04)而THP-1细胞向CEPC的黏附功能显著增强(10.0±2.0和32.0±3.0,P＜0.01);CEPC的迁移功能下降[(4.81±0.23)%和(1.21±0.08)%,P＜0.01];CEPC在Matrigel上的成管功能显著下降(18.0±2.0和12.0±3.0,P=0.024);CEPC中iNOS和IL-6(P均＜0.01)的表达和ICAM-1表达水平均显著增加,而p-akt的表达水平显著下降.结论 活动期SLE患者CEPC数量并未显著减少但其部分功能受损,包括迁移和成管能力等.活动期SLE患者CEPC细胞状态差,抗炎能力降低.     

通心络和他汀类药物对人内皮祖细胞数量与功能的影响

目的:观察中药通心络和他汀类药物对外周血内皮祖细胞(EPCs)数量和功能的影响。方法:采用密度梯度离心法分离培养人外周血EPCs,经FITC-UEA-I和Dil-acLDL双染色鉴定后,并进一步通过流式细胞仪检测其表面标志CD34、CD133,将贴壁细胞随机分为:对照组、阿托伐他汀组(终浓度10μmol/L)、通心络组(终浓度0、50、100、200、500、750和1000μg/ml),作用不同时间(0、12、24、48、60和72h)后,检测细胞形态及计数,再采用四甲基偶氮唑盐微量酶反应比色法(MTT法)、Transwell小室、黏附功能检测评价其增殖、迁移和黏附能力。结果:不同浓度通心络组、阿托伐他汀组均能较对照组明显提高EPCs数量,显著改善其增殖、迁移、黏附能力,通心络在500μg/ml时对细胞数量及功能改善最为显著。采用500μg/ml的通心络进行时效作用的研究,各组呈时间依赖性增强,EPCs的数量和功能在72h达到高峰。结论:通心络和阿托伐他汀均能在体外提高内皮祖细胞的数量及功能。     

不对称二甲基精氨酸对体外培养内皮祖细胞数量和功能的影响

目的:观察不对称二甲基精氨酸(ADMA)对体外培养内皮祖细胞(EPCs)数量和功能的影响。方法:密度梯度离心法获取外周血单个核细胞,培养6d后,收集贴壁细胞并分别加入ADMA1、5、10μmol/L干预培养72h。激光共聚焦显微镜和流式细胞仪鉴定EPCs,分别观察EPCs的数量、增殖及黏附和产生一氧化氮的能力。结果:与对照组相比,不同浓度的ADMA可显著抑制外周血EPCs扩增、黏附增殖能力和产生NO的能力。结论:ADMA可抑制培养EPCs的数量和功能。     

Reduced circulating endothelial progenitor cell number in healthy young adult hyperinsulinemic men

Background and aims

The number of Endothelial Progenitor Cells (EPCs) is considered a novel marker of cardiovascular (CV) disease. It is not clear which are the main determinants of EPC number in apparently healthy subjects in the absence of overt clinical CV or metabolic abnormalities. We evaluated the main clinical determinants of EPC levels in a population of healthy subjects with normal glucose tolerance.

Methods and results

EPC number was determined in 122 healthy subjects (73M/49F;36.6 ± 8yrs). Blood samples were collected to test biochemical variables. OGTT was performed and insulin resistance/compensatory hyperinsulinemia was defined according to fasting plasma insulin (FPI) levels. EPCs were identified as cells co-expressing CD133/CD34/KDR antigens by flow-cytometry. CD133⁺/KDR⁺ count inversely correlated with BMI (rho=−0.18;p < 0.05), waist circumference (−0.2;<0.05), diastolic (−0.23;<0.01) and systolic blood pressure (−0.21;<0.05), uric acid (−0.24;<0.005), PAI-1 (−0.197; <0.05) and FPI (−0.2;<0.05) and directly correlated with HDL cholesterol (0.182;<0.05). CD34⁺/CD133⁺/KDR⁺ count inversely correlated with uric acid (−0.28;<0.005) and FPI (−0.2;<0.05). EPC number was lower in males (p < 0.05) and gender was the only independent predictor of EPC count (p < 0.05). By dividing the population in four subgroups based on gender and insulin resistance, CD133⁺/KDR⁺ levels were lower in insulin resistant compared to insulin sensitive males (p < 0.05) with no differences in females.

Conclusion

The male gender is an independent predictor of low EPC levels in healthy subjects. This might contribute to explaining the higher CV risk in males compared to pre-menopausal age-matched females. In this study a reduced EPC number seems to be associated with insulin resistance in male subjects.     

High irisin levels in overweight/obese children and its positive correlation with metabolic profile,blood pressure,and endothelial progenitor cells

Background and aims

Irisin is involved in the compensatory mechanisms for metabolic regulation and appears to be associated with glucose homeostasis and lipid profile. However, it's possible implications on obesity-associated cardiometabolic complications have not been completely elucidated. This study aimed to investigate the association between irisin level and anthropometric data, metabolic parameters, blood pressure, and endothelial progenitor cells (EPCs) level among children with overweight/obesity.

高同型半胱氨酸血症患者循环内皮祖细胞的变化

目的:观察高同型半胱氨酸血症(Hhcy)患者外周血内皮祖细胞(EPCs)数量和功能的变化.方法:入选患者60例,其中Hhcy患者30例(Hhcy组),非Hhcy患者30例(对照组).采用全自动荧光偏振免疫分析法检测血浆同型半胱氨酸(homocysteine,Hcy),流式细胞仪计数循环EPCs(即AC133 KDR 细胞).此外,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4d后贴壁细胞进行细胞化学分析.激光共聚焦显微镜鉴定FITC标记荆豆凝集素Ⅰ和DiI标记的乙酰化LDL双染色阳性细胞为正在分化的EPCs.采用改良的Boyden小室和黏附能力测定实验观察EPCs的迁移和黏附能力.结果:Hhcy患者循环EPCs数量、外周血正在分化的EPCs数量[(63.9±11.7)、(36.1±6.5)个]均较对照组的[(91.5±14.2)、(51.5±8.3)个]明显减少,均P<0.01;这2种EPCs数量均与血Hcy水平呈反向线性关系.Hhcy患者EPCs的迁移和黏附能力也较对照组的明显受损,EPCs的这2种功能与血Hcy水平呈反向线性关系.结论:Hhcy患者外周血EPCs数量减少、功能减退,且EPCs的数量和功能与血Hcy水平呈反向线性关系.