骨髓刺激对大鼠骨髓源内皮祖细胞的影响  

CHANGES OF ENDOTHELIAL PROGENITOR CELLS IN RATS AFTER BONE-MARROW STIMULATION

作　　者：佟铸[1] 谷涌泉[1] 张建[1] 李建新[1] 张淑文[1] 汪忠镐[1] 

机构地区：[1]首都医科大学宣武医院血管外科血管外科研究所,北京100053

出　　处：《中国修复重建外科杂志》2008年第10期1218-1221,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家高技术研究发展计划(863)资助项目(2006AA02A134);国家自然科学基金资助项目(30471708);北京市自然科学基金资助项目(7072031);北京市委组织部优秀人才资助项目(20061D0501800265)~~

摘　　要：目的观察大鼠骨髓刺激后骨髓源内皮祖细胞(endothelial progenitor cells,EPCs)数量和功能的改变,探讨采用骨髓刺激后骨髓干细胞移植提高下肢缺血性疾病疗效的可能机制。方法取12只SPF级雄性Lewis大鼠,体重200～250g,按是否皮下注射重组人粒细胞集落刺激因子注射液分为刺激组和对照组(n=6)。取骨髓单个核细胞用EBM-2培养液进行诱导分化,培养7d后对贴壁细胞采用异硫氰酸荧光素-荆豆凝血素1和DiI标记的乙酰化低密度脂蛋白染色进行EPCs鉴定及计数;同时采用黏附能力测定实验观察EPCs的黏附能力。另取12只SPF级雄性Lewis大鼠制备单侧后肢缺血模型,在模型制备后3d将8×106个EPCs移植至缺血侧后肢的肌肉内,按照移植EPCs来源不同将后肢缺血大鼠分为刺激组和对照组(n=6)。EPCs移植3周后行后肢血管造影,计算缺血侧后肢侧支血管数目。结果大鼠骨髓单个核细胞培养7d后刺激组EPCs数量为(145.2±37.0)个/HP,大于对照组(95.2±39.4)个/HP(P<0.05);刺激组黏附EPCs数量为(21.8±4.3)个/HP,大于对照组(15.0±5.2)个/HP(P<0.05)。大鼠骨髓源EPCs移植至缺血肢体后3周,刺激组EPCs移植后缺血侧后肢侧支血管数目为(4.2±1.2)个,大于对照组的(2.7±0.8)个(P<0.05)。结论骨髓刺激使骨髓源EPCs数量增加、功能改善,可能是骨髓刺激后骨髓干细胞移植提高下肢缺血性疾病疗效的机制之一。Objective To observe the changes in the number and function of bone marrow-derived endothelial progenitor cells （EPCs） after bone-marrow stimulation, and to investigate the possible mechanism of improving ischemic limb disease after bone-marrow stimulation through autologue bone-marrow stem cell implantation. Methods Twelve male Lewis rats, weighing 200-250 g, were classified into the bone marrow stimulation group （n=6） and the control group （n=6）. In the stimulation group, the bone marrow of each rat was stimulated by injection of recombinant human granulocytemacrophage colony-stimulatory factor. Mononuclear cells were harvested from bone marrow and cultured in EBM-2 medium. After 7-day culture, EPCs were stained by 1, 1-dioctadecyl-3, 3, 3, 3-tetramethylindocarbocyanine-labbled acetylated low density lipoprotein/fluorescein isothiocyanate-ulex europaeus agglutinin 1, and the double positive cells were counted by the fluorescent microscope. The adhesive ability of EPCs was determined by counting the number of re-cultured EPCs. The unilateral ischemia hindlimb model was made with 12 Lewis rats. Three days later, EPCs were transplanted into the ischemic tissues. According to different sources of EPCs, the 12 rats were divided into 2 groups： the stimulation group （n=6） and the control group （n=6）. At 3 weeks after EPCs transplantation, the quantity of the collateral vascular was observed by digital subtraction angiography （DSA）. Results After 7-day culture, the number of EPCs in the stimulation and control groups was （145.2 ± 37.0）/HP and （95.2 ± 39.4）/HP, respectively, and there was significant difference between the two groups （P 〈 0.05）. Meanwhile, the number of adhesive EPCs in the stimulation and control groups was （21.8 _± 4.3）/HP and （15.0 _± 5.2）/HP, respectively, and the difference between the two groups was significant （P 〈 0.05）. At 3 weeks after the EPCs implantation, the number of the collateral vascular was significantly larger in the stimulati

关 键 词：重组人粒细胞集落刺激因子 骨髓刺激 内皮祖细胞 细胞移植 大鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]