COPD患者外周血中EPCs增殖、迁移、黏附及血管形成功能研究  

Function of EPCs proliferation,migration,adhesion,and capacities of late-EPCs vascular tube formation altered in perpherial blood of COPD patients

作　　者：刘笑然[1] 刘杨丽[2] 林耿鹏[2] 黄鑫炎[2] 谭卫平[2] 谢灿茂[2] 

机构地区：[1]海南医学院附属医院急诊科,海南海口570102 [2]中山大学附属第一医院呼吸内科,广东广州510080

出　　处：《海南医学院学报》2014年第11期1463-1468,共6页Journal of Hainan Medical University

基　　金：国家自然科学基金资助项目(81260010);海南省自然科学基金资助项目(309064)~~

摘　　要：目的:观察慢性阻塞性肺病(COPD)患者外周血中内皮祖细胞(EPCs)增殖、迁移、黏附及小管形成等功能。方法:体外培养分离COPD患者及对照组外周血中EPCs,采用MTT测定细胞不同时间段的OD值,观察EPCs增殖情况。应用Transwell测定细胞对基质细胞衍生因子-1(SDF-1)的迁移能力。通过对细胞HUVEC的黏附及late-EPCs小管形成实验判断COPD患者外周血中EPCs对血管内皮的修复能力,并用NOS/SCID下肢股动脉内皮损伤模型观察患者EPCs修复功能。结果:COPD患者的EPCs和对照组单个集落细胞数量分别是22.0±7.0、62.0±12.9(100倍,10个随机视野),两者存在明显差异(P<0.001)。细胞迁移实验中发现,COPD患者组early-EPCs对SDF-1迁移细胞数量明显少于对照组,两组迁移的细胞数量分别是0.425×104、0.775×104(P=0.039),通过流式分析发现COPD患者组early-EPCs表达C-X-C家族趋化因子受体4(CXCR4)数量较对照组少,两者分别是(55.1±9.9)%、(91.5±6.7)%。Late-EPCs对HUVECs形成小管黏附实验中,两者有明显差异(P<0.001),分别是26.4±8.1、35.2±7.3/×10(10随机视野)。小管成形实验中,COPD患者的late-EPCs不能形成完整的小管;动物在体实验中发现,COPD患者组late-EPCs在受损血管处黏附较对照组少。结论:COPD患者外周血中EPCs增殖、迁移、黏附等功能降低,血管形成功能异常。Objective： To determine the role of early EPCs in COPD patients in terms of migration by SDF-1, proliferation, adhesion, and capacities of late-EPCs vascular tube formation which may contribute to the pathogenesis of CVD in patients with COPD. Methods： EPCs from 25 COPD and 16 control patients were isolated by Ficoll density-gradient centrifugation and identified using fluorescence activated cell sorting. Transwell Migratory Assay was performed to determine the number of EPCs colony forming units and the adherent capacity late-EPCs to human umbilical vein endothelial cells. Following arterial damage in NOD/SCID mice, the number of EPCs incorporated at the injured vascular site was determined using a fluorescence microscope. It showed that the number of EPCs clusters and cell migration, as well as the expression of CXCR4, was significantly decreased in COPD patients. Additionally, the number of late-EPCs adherent to HUVEC tubules was significantly reduced and fewer VEGFR2＋-staining cells were incorporated into the injured site in COPD patients. Results： EPCs clonal units from COPD patients had reduced proliferation rates, and they were smaller, with a reduced number of clusters compared to control patients. In the transwell lower chamber, the numbers of early-EPCs isolated from patients with COPD were significantly less than that of control subjects at increasing concentrations of SDF-la （P〈0.05）. Fewer COPD late-EPCs were incorporated into tubules compared with control EPCs （26.4±8.1 versus 35.2±7.3, respectively, × 10 power, 5 fields, P〈 0. 001）. Angiogenesis assays showed that the formation of vascular networks in a 24-well coated plate with matrigel was markedly impaired in the COPD patients compared to control subjects. The injured arterial slices were observed under a fluorescence microscope, The Fluorescence density of late-EPCs from COPD patients at the site of injury site were less than control subjects. Conclusions： Our data showed that human endothelial progenitor cells isolated

关 键 词：慢性阻塞性肺病 心血管疾病 内皮祖细胞 C-X-C家族趋化因子受体4 

分 类 号：R563[医药卫生—呼吸系统]