慢病毒载体介导的CXCR4基因过表达对骨髓间充质干细胞归巢影响  

Effect of Lentiviral Vector-Mediated CXCR4 Gene Overexpression on Mesenchymal Stem Cell Homing Capacity

作　　者：陈伟 周典 李亚茹 刘俊秀 王祥民 朱峰 徐开林 CHEN Wei;ZHOU Dian;LI Ya-Ru;LIU Jun-Xiu;WANG Xiang-Min;ZHU Feng;XU Kai-Lin(Department of Hematology,The Affiliated Hospital of Xuzhou Medical University,Xuzhou,221002,Jiangsu Province,China)

机构地区：[1]徐州医科大学附属医院血液科,江苏徐州221002

出　　处：《中国实验血液学杂志》2018年第5期1543-1547,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金(81300441);江苏省自然科学基金(BK20161177);中国博士后基金(2016M590507);江苏省卫生国际交流支撑计划(JSH-2017-008)

摘　　要：目的:研究小鼠CXCR4基因过表达对小鼠骨髓间充质干细胞(mesenchymal stem cell, MSC)体内归巢特性的影响。方法:利用慢病毒载体介导小鼠MSC过表达CXCR4基因,建立过表达CXCR4的MSC。将BALB/c小鼠分为单纯照射组(TBI组):受鼠接受全身照射(total body irradiation, TBI)后输注生理盐水;受鼠TBI后经鼠尾静脉输注EGFP空载体转导的MSC 5×10~5空载体对照组(EGFP-MSC)和受鼠TBI后经鼠尾静脉输注同时携带EGFP和CXCR 4基因的MSC 5×10~5过表达CXCR4组(CXCR4-MSC)3组,每组动物10只。冰冻切片观察MSC在受鼠体内重要脏器的分布,应用流式细胞术检测MSC归巢至受鼠体内骨髓及脾脏效率,酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)检测受鼠外周血和骨髓中基质细胞衍生因子1(stromal cell derived factor-1, SDF-1)水平。结果:冰冻切片结果显示,过表达CXCR 4可明显提高MSC归巢至肺脏、肝脏和脾脏的效率;流式细胞术测定过表达CXCR4的MSC归巢至脾脏和骨髓数量明显高于EGFP对照组(P <0.05);ELISA结果显示照射24小时后小鼠外周血及骨髓中SDF-1水平明显升高,并且SDF-升高水平与过表达CXCR4的MSC归巢效率存在相关性。结论:利用慢病毒载体介导的过表达CXCR4基因可促进小鼠骨髓间充质干细胞归巢。Objective:To explore the effects of lentiviral-mediated CXC chemokine receptor-4(CXCR-4)gene over-expression on the homing capacity of mesenchymal stem cells(MSC)in vivo.Methods:The MSC overexpressing CXCR-4 were constructed by using the lentiviral vector-mediated mouse MSC overexpressing the CXCR-4 gene.The BALB/c mice were divided into 3 group:simple radiation group(TBI)in which mice exposed to total body irradiation,then were infused with normal saline;EGFP-MSC group in which mice were infused with MSC(5×105)transducted by EGFP via tail vein after TBI;and CXCR-4-MSC group in which mice were infused with MSC(5×105)simultaneously carraying EGFP and CXCR-4 gene via tail vein after TBI.The mice were sacrified at 24 hours after infusion,the frozen sections were prepared to detect the distribution of infused MSC.Furthermore,the numbers of MSC homing into spleen and bone marrow was detected by flow cytometry,and the level of stromal cell-derived factor-1(SDF-1)was detected by ELISA.Results:The frozen section showed that the CXCR-4 over-expression could significantly enhance the efficacy of MSC homing into lung,liver and spleen;the flow cytonetry detection slowed that the number of over-expressed CXCR-4 MSC homing into spleen and bone matrow was sigmificantly higher than that in EGFP-MSC group(P<0.05),the ELISA showed that the SDF-1 level in peripheral blood and bone marrow after 24 hours of irradiation significantly incrtaoed(P<0.05),moreover,the SDF-1 level increase was associcted with horming efficacy of MSC with CXCR-4 overexpression.Conclusion:overexpression CXCR-4 gene mediated by lentiviral vector can prmote the efficacy of MSC homing into spleen and bone marrow.

关 键 词：C-X-C型趋化因子受体4 间充质干细胞 慢病毒载体 归巢 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]