血管生成素1在G-CSF诱导的造血干/祖细胞动员中的表达变化  

Changes of angiopoietin 1 expression in G-CSF induced hematopoietic stem progenitor cells mobilization

作　　者：李天寿[1] 李桥川[1] 李斯丹[2] 聂胤超 邱录贵[3] 

机构地区：[1]广西医科大学第一附属医院,南宁530021 [2]儿童血液病与肿瘤分子分型北京市重点实验室,儿科重大疾病研究教育部重点实验室,首都医科大学附属北京儿童医院血液肿瘤中心 [3]中国医学科学院、北京协和医学院血液学研究所、血液病医院

出　　处：《中华血液学杂志》2015年第5期418-421,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金（81160075、81300433）;广西自然科学基金（0728124）;北京市自然科学基金（7144211）

摘　　要：目的 探讨G-CSF诱导的造血干/祖细胞（HSPC）动员过程中血管生成素1（Ang1）在小鼠骨髓的表达变化及导致其变化的机制.方法 采用流式细胞术检测小鼠动员前及动员后外周血Lin Scal＋cKit＋（LSK）细胞比例变化,并应用ELISA、免疫组化、RQ-PCR等方法检测G-CSF动员不同时间点小鼠外周血和骨髓标本,观察Ang1、骨钙素（OCN）基因及蛋白水平变化,显微镜下计数骨髓标本成骨细胞数量.结果 G-CSF动员5d小鼠外周血LSK细胞比例为（0.61±0.05）％,相对于对照组的（0.04±0.01）％显著增高（P＜0.05）;稳态小鼠OCN及Ang1 mRNA在骨内膜细胞表达高于骨髓有核细胞;动员后骨内膜细胞OCN及Ang1 mRNA相对表达量对照组（28.64±8.61及2.84±0.95）、G-CSF动员3d组（12.55±7.06及0.93±0.30）及动员5d组（4.75±1.62及0.92±0.22）均下降;动员后骨内膜成骨细胞数量显著减少,骨髓组织Ang1蛋白表达下降;对照组、动员3d及5d组外周血OCN及Ang1浓度分别为（24.11±3.17）及（2.24±0.52）ng/ml、（9.96±2.16）及（1.21±0.38）ng/ml和（8.43±2.62）及（0.90±0.24）ng/ml,动员3d及5d组均明显低于对照组,差异均有统计学意义（P值均＜0.05）.结论 在G-CSF介导的HSPC动员过程中,骨髓微环境中成骨细胞数量下降、功能减低,进而导致骨髓Ang1表达下降,导致骨髓对HSPC的趋化和黏附能力下降,有利于HSPC动员的发生.Objective To investigate the changes and mechanism of angiopoietinl （Ang1） in murine bone marrow during G-CSFinduced mobilization of hematopoietic stem/progenitor cell.Methods The proportion of Lin-Sca1＋cKit＋ （LSK） cells in peripheral blood of C57BL/6 mice before and after G-CSF mobilization was detected by flow cytometry.Expression changes of Ang1 and osteocalcin （OCN） during HSC mobilization were determined by immunohistochemistry,enzyme linked immunosorbent assay （ELISA） and real-time fluorescence quantitative PCR.The number of osteoblasts in the bone marrow was counted under the microscope.Results After treated with G-CSF,the proportion of LSK cells in peripheral blood significantly increased from the controls （0.04±0.01）% to （0.61±0.05）% at day 5 （P〈 0.05）.Before G-CSF mobilization,the endosteum cells expressed higher level of OCN and Ang1 than that of bone marrow nucleated cells.The mRNA expression level of OCN was significantly reduced from 28.64±8.61 in the controls to 12.55±7.06 on day 3 and 4.75±1.62 on day 5,and the expression level of Ang1 also declined from 2.84±0.95 in the controls to 0.93±0.30 on day 3 and to 0.92±0.22 on day 5 after G-CSF mobilization.The number of endosteum osteoblasts was significantly decreased after mobilization （P〈 0.05）.The Ang1 expression was decreased in the BM after mobilization.The serum OCN was significantly reduced from （24.11±3.17） ng/ml in the controls to （9.96±2.16） ng/ml on day 3 and （8.43±2.62） ng/ml on day 5,and the Ang1 also declined from （2.24±0.52） ng/ml in the controls to （1.21±0.38） ng/ml on day 3 and （0.90±0.24） ng/ml on day 5.Conclusion In G-CSFinduced HSPC mobilization,the bone marrow osteoblasts retraction causes reduction of Angl,and the reduction of Angl may contribute to HSPC mobilization.

关 键 词：血管生成素1 成骨细胞 造血干细胞动员 Anggiopoitin-1 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]