氯膦酸二钠脂质体清除巨噬细胞对小鼠造血干祖细胞归巢过程的影响  

The effect of macrophages depletion by clodronate liposome on homing of hematopoietic stem/progenitor cells

作　　者：杨骁 杨欣[2] 张晓龙 YANG Xiao;YANG Xin;ZHANG Xiao-long(Core Research Laboratory, the Second Affiliated Hospital of Xi′an Jiaotong University, Shaanxi Province, Xi′an 710004, China;Department of Rheumatism and Immunology, the Second Affiliated Hospital of Xi′an Jiaotong University, Shaanxi Province, Xi′an 710004, China;Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin 300060, China)

机构地区：[1]西安交通大学第二附属医院科研中心实验室,陕西西安710004 [2]西安交通大学第二附属医院风湿免疫科,陕西西安710004 [3]天津医科大学肿瘤医院国家肿瘤临床医学研究中心,天津300060

出　　处：《河北医科大学学报》2022年第2期135-139,192,共6页Journal of Hebei Medical University

基　　金：国家自然科学基金(81802862);陕西省自然科学基础研究计划(2020JQ-546)。

摘　　要：目的探讨清除巨噬细胞对小鼠造血干祖细胞(hematopoietic stem/progenitor cells,HSPC)归巢过程的影响。方法免疫磁珠分选获得小鼠HSPC,使用羧基荧光素二醋酸盐琥珀酰亚胺酯(Carboxyfluorescein diacetate succinimide ester,CSFE)染色标记,采用氯膦酸二钠脂质体(clodronate liposomes,Clod)方法建立去除巨噬细胞的小鼠模型,通过流式细胞术检测HSPC移植的归巢效率,通过实时荧光定量逆转录酶链式反应方法和酶联免疫吸附测定方法检测去除巨噬细胞后骨髓微环境中CXCL12、SCF和人血管内皮细胞粘附分子1(vascular adhesion molecule 1,VCAM1)分子mRNA表达水平和蛋白表达水平的变化。结果氯膦酸二钠脂质体作用后第24小时和第48小时,小鼠骨髓中CD11b+F4/80+巨噬细胞数量显著减少(P<0.05);移植小鼠HSPC后,相较于对照组,去除巨噬细胞组小鼠HSPC归巢效率显著降低(P<0.05);同时去除巨噬细胞组小鼠骨髓微环境中CXCL12、SCF的mRNA和蛋白表达量比对照组显著降低(P<0.05),而VCAM1表达量无显著变化。结论去除巨噬细胞能够显著降低小鼠HSPC归巢效率,该作用可能与去除巨噬细胞后导致的骨髓微环境中CXCL12与SCF含量减少有关。Objective To explore the effect of macrophage depletion on the homing process of hematopoietic stem/progenitor cells(HSPC).Methods Mouse HSPCs were obtained by immunomagnetic bead sorting,and labeled with carboxyfluorescein diacetate succinimide ester(CSFE).A mouse model of macrophage depletion was established by clodronate liposomes(Clod),and the efficiency of HPSC homing was detected by flow cytometry.The mRNA and protein expression levels of CXCL12,SCF and vascular adhesion molecule 1(VCAM1)molecules in bone marrow niche after macrophage depletion were detected by real-time RT polymerase chain reaction(qRT-PCR)and enzyme linked immuno sorbent assay(ELISA)assay.Results The number of CD11b+F4/80+macrophages in the bone marrow of mice was significantly decreased at the 24 and 48 h after the injection of Clod(P<0.05).After transplantation of mouse HSPC,the homing efficiency of HSPC in the mice in the Clod group was significantly reduced,as compared with the control group(P<0.05).In the meantime,the mRNA and protein expressions of CXCL12 and SCF in the bone marrow niche of the mice in the Clod group were significantly lower than those in the control group(P<0.05),while there was no significant change in VCAM1 expression between Clod and control group.Conclusion Macrophage depletion can reduce the homing efficiency of HSPC,which may be related to the reduction of CXCL12 and SCF levels in the bone marrow niche after macrophage depletion.

关 键 词：骨髓祖代细胞 巨噬细胞 氯膦酸二钠脂质体 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]