小鼠异基因骨髓移植γ线清髓性照射对骨髓内皮龛的损伤研究  

Injury of Bone Marrow Endothelial Niche by Irradiation Myeloablative Conditioning in Mouse Allo-BMT

作　　者：汤仁仙[1] 丁爽[2] 徐开林[2] 贾路[2] 闫志凌[2] 陈翀[2] 曾令宇[2] 

机构地区：[1]徐州医学院病原生物教研室,江苏徐州221002 [2]徐州医学院附属医院血液科,江苏徐州221002

出　　处：《中国实验血液学杂志》2010年第6期1579-1584,共6页Journal of Experimental Hematology

基　　金：江苏省自然科学基金(BK2007504);江苏省"六大人才高峰"项目(09-B1-033);江苏省"青蓝工程"

摘　　要：本研究探索异基因骨髓移植清髓性γ线照射预处理对受鼠骨髓内皮的损伤程度。体外培养小鼠骨髓单个核细胞,经5-7天检测其表面标记、吞噬Dil-Ac-LDL和结合FITC-UEA-1鉴定,并行CFSE标记。分析正常组、清髓性照射组、内皮祖细胞(endothelial progenitor cells,EPC)移植组及照射联合EPC移植组小鼠外周血中白细胞变化、骨髓内皮的改变及CFSE标记EPC的骨髓内分布。结果发现,培养细胞鉴定证实为CD31+CD133+CD45low/-,且具有吞噬Dil-Ac-LDL和结合FITC-UEA-1能力。小鼠清髓照射后外周血白细胞迅速减少,与正常组相比,有显著差异(p<0.05)。照射后3天,骨髓中大量出血,内皮细胞和基底膜间连接被损坏。清髓照射后输注CFSE标记EPC,18小时后小鼠骨髓中可见CFSE+细胞,其细胞量是正常小鼠单纯EPC输注组的58倍(p<0.05)。结论:移植清髓照射预处理引起严重骨髓内皮龛损伤,该损伤驱动外源性EPC的归巢。The aim of study was to investigate the injury of bone marrow microenvironment after γ ray irradiation conditioning in mouse allogeneic hematopoietic stem cell transplantation （allo-HSCT）. The mononuclear cells collected from mice bone marrow for culture in vitro, were identified by flow cytometry with double staining when cultured for 5-7 days. Mice were separated randomly into 4 groups, namely, the control group, irradiation group, endothelial progenitor cell（EPC） transplantation group and irradiation combined EPC transplantation group. Peripheral blood was collected to assay the circulating white blood cells. The histological, electron microscopic and immunofluorescence analyses of bone marrow were performed in the same time, furthermore the distribution of labeled EPC was determined. The results showed that EPC were identified as CD45low/-CD133＋CD31＋, double positive of Dil-Ac-LDL and FITC-UEA-1. The bone marrow microenvironment injury of recipient mice was shown in the irradiation group in which the number of WBC began to decrease after conditioning, and the mice were all died at 8 days （p0.05）. The intramedullary hemorrhage could be detected by light microscopy at 3 days after irradiation, when the destruction of connection between endothelial cell and the basement membrane was observed by TEM. There were CFSE labeled cells in bone marrow in irradiation combined EPC transplantation group at 18 hours after transplanted cultured EPC in vitro, the number of CFSE＋ cells was 58 -folds of EPC transplantation group （p0.05）. It is concluded that the irradiation can cause the severe endothelium injury that drives extrinsic EPC homing to the injured bone marrow microenvironment.

关 键 词：异基因骨髓移植 Γ线照射 骨髓内皮龛 

分 类 号：R457.7[医药卫生—治疗学] R818[医药卫生—临床医学]