流体剪切应力对晚期内皮祖细胞生物学功能的影响  

Effects of shear stress on late endothelial progenitor cell functions

作　　者：成敏[1] 尹青令[1] 崔晓栋[1] 张晓芸[1] 李鑫[1] 李宏[1] 官秀梅[1] 王建英[1] 

机构地区：[1]潍坊医学院基础医学院,山东潍坊261053

出　　处：《医用生物力学》2014年第1期60-65,共6页Journal of Medical Biomechanics

基　　金：国家自然科学基金资助项目(30900290;31270993);国家教育部"新世纪优秀人才计划"资助项目(NCET-10-0922);山东省自然科学基金资助项目(ZR2011CQ030);山东省医药卫生科技发展计划(2011QZ024)

摘　　要：目的研究流体剪切应力处理对晚期内皮祖细胞(endothelial progenitor cells,EPCs)体外及体内生物学功能的影响。方法密度梯度离心法分离大鼠骨髓单核细胞,应用EGM-2MV进行体外培养。以3~4代的EPCs,即晚期EPCs为靶细胞,对其施以1.2 Pa剪切应力处理。采用EdU标记技术、黏附能力测定实验、改良的Boyden小室、Annexin V/PI、β-半乳糖苷酶检测法、Matrigel法、荧光定量RT-PCR等方法分别检测剪切应力对晚期EPCs增殖、黏附、迁移、凋亡、衰老、体外成血管及VEGF mRNA表达等生物学功能的影响。应用大鼠颈动脉损伤模型及细胞原位移植等实验手段检测剪切应力预处理对晚期EPCs修复受损内皮的影响。结果 1.2 Pa剪切应力处理可不同程度提高晚期EPCs的增殖、黏附、迁移及成血管能力(P<0.01),上调VEGF的基因表达,抑制晚期EPCs的衰老及凋亡(P<0.01);移植经剪切应力预处理的晚期EPCs可加速损伤内皮的修复,减缓内膜的增生。结论流体剪切应力可改善晚期EPCs的功能活性,提高晚期EPCs修复损伤血管内皮的能力,这为EPCs的临床应用及剪切应力介导的细胞疗法提供了实验依据。Objective To investigate the effects of shear stress on late endothelial progenitor cells （EPCs） functions in vitro and in vivo. Methods Density gradient centrifugation-isolated rat bone marrow mononuclear cells were cultured in EGM-2MV and induced into EPCs. The 3rd^4th generation of EPCs, namely late EPCs, were treated with shear stress （1.2 Pa）. Then cell biological functions, such as proliferation, adhesion, migration and ability of tube formation, were assayed with EdU incorporation assay, adhesion testing, Boyden chamber assay and Matrigel, respectively. The gene expression of VEFG was analyzed by real time RT-PCR. The apoptosis and aging situation of late EPCs were assayed by FACS and senescence-associated β-galactosidase （SA-β-gal） staining. The reendothelialization capacity of late EPCs treated by shear stress was evaluated by establishing models of freshly balloon-injured carotid arteries of rats and cell transplantation in situ. Results Shear stress could increase proliferation, adhesion, migration and tube formation of late EPCs （P〈0.05）, upregulate the gene expression of VEGF, inhibit EPC apoptosis and delayed EPC aging （P〈0.05）. Transplantation of late EPCs treated by shear stress facilitated in vivo reendothelialization in the injured arterial segment and inhibited neointima formation. Conclusions Shear stress within the physiological range can improve the functions of late EPCs and enhance their therapeutic ability of repairing vascular endothelial injury, which provides experimental basis for the clinic application of EPCs and shear stress-mediated cell therapy.

关 键 词：剪切应力 内皮祖细胞 细胞黏附 再内皮化 

分 类 号：R593.2[医药卫生—内科学]