缺氧预处理外周间充质干细胞对兔血管成形术后再狭窄的影响  

Effects of transplantation of peripheral blood mesenchymal stem cells with hypoxia preconditioning on postangioplasty restenosis in rabbits

作　　者：石蓓[1] 郭艳[1] 王正龙[1] 王冬梅[1] 沈长银[1] 

机构地区：[1]遵义医学院第一附属医院心血管内科,贵州遵义563003

出　　处：《中国病理生理杂志》2009年第9期1686-1691,共6页Chinese Journal of Pathophysiology

基　　金：贵州省教育厅重点项目(No.黔教科2007-040号)

摘　　要：目的:探讨经骨髓动员后外周血分离的间充质干细胞(MSCs)移植对动脉粥样硬化模型兔颈动脉球囊成形术后再狭窄的防治作用及可能机制。方法:粒细胞集落刺激因子(G-CSF)动员、外周血分离体外扩增获得MSCs,增强型绿色荧光蛋白(EGFP)报告基因标记备用。建立兔动脉粥样硬化狭窄模型,随机分为缺氧预处理MSCs移植组(HP)、非缺氧预处理MSCs移植组(NHP)及对照组(C),再于球囊成形术后立即耳缘静脉内分别注入标记的缺氧处理、非处理MSCs或培养液。术后7 d、14 d、28 d采集外周血用ELISA法检测血管内皮生长因子(VEGF)水平;术后28 d处死动物留取血管标本通过Weigert弹力纤维染色及免疫组化染色进行形态学分析、归巢MSCs分析、再内皮化程度分析。结果:HP组、NHP组术后各时点VEGF水平均显著高于C组(P<0.01);术后7d、14 d HP组显著高于NHP组(P<0.05),28 d则差异无显著。术后7 d血管组织中仅HP组、NHP组血管组织中有EGFP(+)细胞分布;术后28 d HP组内膜增殖程度、再狭窄率明显低于NHP组(P<0.05),而该2组均显著低于C组(P<0.01);再内皮化程度HP组明显优于NHP组(P<0.05)、同时该2组又均显著优于C组(P<0.01)。结论:MSCs移植能够抑制模型兔颈动脉球囊成形术后内膜增生、促进损伤血管快速内皮化从而降低再狭窄率,移植经缺氧预处理的MSCs可增强上述作用,可能与缺氧应激诱导MSCs分泌细胞因子功能增强有关。AIM： To investigate the function and the mechanism of transplanting bone marrow derived peripheral blood mesenchymal stem cells （PBMSCs） on restenosis after carotid balloon angioplasty in the model of carotid athero- sclerosis rabbits, and to determine if the functions of PBMSCs are enhanced after hypoxia preconditioning. METHODS： Bone marrow cells were mobilized by granulocyte colony - stimulating factor （ G - CSF）, and PBMSCs were collected through density gradient centrifugation and adherent culture, labeled with enhancement type green fluorescent protein （EG- FP） genes. All animals with carotid atherosclerosis stenosis were randomly divided into three groups： hypoxia preconditioning group （ n = 24, received intravenous transplantation of PBMSCs with hypoxia preconditioning）, non - hypoxia precondi- tioning group （n = 24, received normal culture of PBMSCs） and control group （ n = 24, only received equal - volume of euhure medium）. Vascular endothelial growth factor （VEGF） was determined by enzyme linked immunosorbent assay （ELISA） at 7 d, 14 d and 28 d post -angioplasty, respectively. The vessel morphology, the homing of MSCs and the reendothelialization were analyzed with Weigert staining and immunohistochemistry. RESULTS： Compared to control group, the level of VEGF significantly increased in both hypoxia preconditioning group and non - hypoxia preconditioning group at all time points （P 〈 0.01 ）. The level of VEGF in hypoxia preconditioning group was higher than that in non - hypoxia pre- conditioning group （P 〈0. 05） at 7 d and 14 d, but no difference at 28 d post - angioplasty was observed. At 7 d, GFP - positive cells were found both in hypoxia preconditioning group and non - hypoxia preconditioning group. Neointima thickening and the rate of restenosis were lower in hypoxia preconditioning group than those in non - hypoxia preconditioning group at 28 d （ P 〈 0. 05 ）, but both hypoxia preconditioning group and non - hypoxia preconditioning group w

关 键 词：间充质干细胞 缺氧 兔 绿色荧光蛋白 再内皮化 再狭窄 

分 类 号：R543.5[医药卫生—心血管疾病]