过表达CXCR4促进小鼠骨髓间充质干细胞的归巢和增殖  

Over-expression of CXCR4 promotes homing and proliferation of mouse bone marrow mesenchymal stem cells

作　　者：王志红[1] 林芸[1] 叶海燕 陈为民[1] 尚晋[1] 魏天南[1] WANG Zhihong;UN Yun;YE Haiyan;CHEN Weimin;SHANG Jin;WEI Tiannan(Department of Hematology,Fujian Provincial Hospital,College of Clinical Medicine,Fujian Medical University,Fuzhou 350001,China)

机构地区：[1]福建医科大学省立临床医学院福建省立医院血液科

出　　处：《细胞与分子免疫学杂志》2019年第5期393-398,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：福建省卫生计中青年骨干人才培养项目(2016-ZQN-4);福建省科技创新联合基金项目(2017Y9068)

摘　　要：目的研究过表达CXC趋化因子受体4(CXCR4)对小鼠骨髓间充质干细胞(BMMSC)体内外归巢特性和细胞增殖活性的影响。方法利用慢病毒载体介导小鼠BMMSC过表达CXCR4,构建过表达CXCR4的细胞(CXCR4-BMMSC)。TranswellTM小室实验检测基质细胞衍生因子1(SDF-1)对BMMSC迁移能力的影响;噻唑蓝(MTT)法检测BMMSC的增殖能力。BALB/c小鼠接受全身照射(TBI)后,随机分为2组,每组10只。BMMSC对照组:小鼠经TBI后,输注5×10^5个绿色荧光蛋白标记的BMMSC(GFP-BMMSC);CXCR4-BMMSC组:小鼠经TBI后,回输5×10^5个携带GFP的CXCR4-BMMSC。尾静脉回输细胞后,取小鼠胸腺进行冰冻切片,荧光显微镜下观察回输的细胞归巢至胸腺组织的情况;流式细胞术检测BMMSC归巢至受鼠体内胸腺的效率。结果 TranswellTM小室实验证实CXCR4可促进SDF诱导的BMMSC的跨膜迁移;与BMMSC对照组相比,CXCR4-BMMSC的增殖活性显著升高。与BMMSC对照组相比,过表达CXCR4可明显提高BMMSC归巢至胸腺组织的效率;流式细胞术检测结果提示BMMSC归巢至胸腺数量高于对照组。结论过表达CXCR4可增强SDF-1对BMMSC的迁移募集,体内可促进BMMSC归巢至损伤胸腺。CXCR4还可促进小鼠BMMSC的增殖。Objective To explore the effects of over-expression of C-X-C motif chemokine receptor 4(CXCR4) on the homing capacity of bone marrow mesenchymal stem cells(BMMSCs) in vivo and in vitro, and the proliferation activity of BMMSCs. Methods The CXCR4-BMMSCs were constructed by the lentiviral vector-mediated BMMSC over-expressing the CXCR4 gene. The effect of chemokine SDF-1 on the migration ability of BMMSCs was detected by TranswellTM migration assay;and MTT assay was used to detect cell proliferation. The BALB/c mice were randomly divided into two groups after receiving total body irradiation(TBI), 10 mice in each group. BMMSCs control group: mice were infused with BMMSCs(5×10^5) transducted by EGFP via tail vein after TBI;CXCR4-BMMSC group: mice were infused with BMMSCs(5×10^5) simultaneously transducted by GFP and CXCR4 gene via tail vein after TBI. The thymus of mice was frozen and sectioned, and fluorescence microscopy was used to observe the homing of BMMSCs into thymus tissue. Furthermore, flow cytometry was performed to detect the homing efficiency of BMMSCs to thymus in recipient mice. Results Transwell assay showed that over-expresion of CXCR4 could promote the migration and recruitment of BMMSCs to chemokine SDF-1;the proliferation activity of CXCR4-BMMSCs increased significantly compared with BMMSC control group. Compared with the control group, the frozen section showed that over-expression of CXCR4 could significantly improve the efficiency of BMMSC homing to thymus tissue. Flow cytometry suggested that CXCR4-BMMSCs homing to thymus were more than those in the control group. Conclusion Over-expression of CXCR4 gene mediated by lentivirus vector can enhance the migration and recruitment of BMMSCs by SDF-1, and promote BMMSCs homing to damaged thymus in vivo. In addition, CXCR4 also promotes the proliferation of BMMSCs.

关 键 词：CXC趋化因子受体4(CXCR4) 基质细胞衍生因子1(SDF-1) 骨髓间充质干细胞(BMMSC) 过表达基因 移植 归巢 

分 类 号：R965[医药卫生—药理学] Q253[医药卫生—药学]