【摘要】 目的探讨兔颈动脉粥样硬化狭窄血管成形术后,骨髓间充质干细胞(BMSC)移植对损伤血管内皮修复的影响及可能机制。方法制作兔颈动脉粥样硬化狭窄模型48只,随机分成BMSC移植组(n=24)和对照组(n=24)。体外培养兔BMSC,流式细胞仪鉴定BMSC表面标志,DAPI标记后备用。球囊损伤颈动脉的即刻,BMSC移植组以107/kg的细胞数经颈外动脉移植到损伤动脉局部,对照组注射等量的PBS液。术前及细胞移植后3、7、14及28天采集外周血用ELISA法测定血管内皮生长因子水平。移植后7天取材观察DAPI标记BMSC的归巢。移植后14天免疫组织化学染色检测损伤血管组织的血小板-内皮细胞黏附分子(CD31)的表达;移植后28天HE染色后测定损伤血管新生内膜面积、新生内膜面积/中膜面积及再狭窄率。结果 BMSC移植组在移植后各时间点血管内皮生长因子水平均显著高于对照组(P<0.01)。移植后7天BMSC移植组损伤血管的内膜表面见DAPI标记的BMSC。移植后14天血管内膜有CD31的连续性表达,对照组没有表达。与对照组比较,移植后28天BMSC移植组血管新生内膜面积(0.092±0.009比0.189±0.007,P<0.01)、新生内膜面积/中膜面积(0.698±1.570比1.630±0.122,P<0.01)及再狭窄率(41.70%±3.70%比61.28%±1.57%,P<0.01)均显著减轻。结论局部移植BMSC可促进粥样硬化狭窄颈动脉球囊损伤后的内皮修复,从而有助于减轻血管再狭窄。更多还原

【Abstract】 Aim To investigate the effect and possible mechanism of bone marrow mesenchymal stem cells(BMSC)transplation on reparing injured vessels endothelium after carotid atherosclerosis stenosis angioplasty in rabbits.Methods Carotid atherosclemsis stenosis model of 48 rabbits had been successfully built up and randomly divided into BMSC transplantation group(n=24) and control group(n=24).BMSC were obtained by density gradient centrifugation and adherent cultured.Surface markers of BMSC were detected by flow cytometry,and BMSC pre-labled by DAPI.Balloon injured carotid artery of rabbits,meanwhile,BMSC(107/kg)were infused into injured artery of BMSC transplantation group rabbits by external carotid artery,and control group infused the same amount of PBS solution.The peripheral blood was collected and the vascular endothelial growth factor(VEGF) levels was detected by enzyme linked immunosorbent assay(ELISA) at preoperative and 3,7,14,28 days of BMSC transplantation.7 days after BMSC transplantation,DAPI labeled BMSC were detected by immunofluorescence microscopy.14 days after BMSC transplantation,the immunohistochemical staining was used to analyse platelet-endothelial cell adhesion molecule(CD31)expression in the injured vessels.28 days after BMSC transplantation,the neointimal area,the ratio of the intima/media area and vascular restenosis were analysed in the vascular tissue by hematoxylin and eosin staining.Results The expression of VEGF in BMSC transplantation group were elevated significantly compared with control group at 3,7,14,28 days after BMSC transplantation.7 days after BMSC transplantation,DAPI labeled BMSC were detected in injured vessels intima.14 days after BMSC transplantation,CD31 continues to express in intima of BMSC transplantation group,while the control group did not express CD31.The neointimal area(0.092 ± 0.009 vs 0.189 ± 0.007,P<0.01),the ratio of the intima/media area(0.698 ± 1.570 vs 1.630 ± 0.122,P<0.01) and the luminal stenosis ratio(41.70% ± 3.70% vs 61.28% ± 1.57%,P<0.01) were significantly decreased in BMSC transplantation group compared with control group at 28 days.Conclusion BMSC transplantation can promote repairing of endothelial after atherosclerotic stenosis carotid artery by balloon injury and reduce the restenosis of injured vessels.更多还原