人骨髓间充质干细胞在mdx鼠体内的动态分布  

Dynamic Distribution of Implanted Human Bone Marrow Mesenchymal Stem Cells in mdx Mice

作　　者：柳太云[1] 冯善伟[2] 李才明[2] 曾缨[2] 姚晓黎[2] 黄文[2] 张成[2] 

机构地区：[1]贵州省人民医院神经内科,贵阳550002 [2]中山大学第一附属医院神经内科,广州510080

出　　处：《中国医学科学院学报》2008年第2期191-196,I0003,I0004,共8页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(30370510);教育部博士点基金(20030558058);贵州省科技基金([2007]2116)~~

摘　　要：目的探讨人骨髓间充质干细胞（hBM-MSCs）在mdx鼠体内的动态分布特点。方法用5．溴脱氧尿嘧啶核苷（Brdu）标记hBM-MSCs，将1×10^7个干细胞（0．3ml）经尾静脉注入免疫抑制的mdx鼠（8—10周龄）体内。于移植后48h、2、4、8、12、16、20、24周处死鼠，用免疫荧光法检测各组织器官的BrdU阳性细胞核数、骨骼肌人细胞核（Hu）及人抗肌萎缩蛋白（Dys）的表达，RT-PCR检测人DysmRNA的表达。结果移植后4周内，BrdU阳性细胞核可见于多数组织、器官，其中以骨髓、肝、肺较多；肝、肺Brdu阳性细胞核数4周后逐渐下降；移植后48h骨髓BrdU阳性细胞核数较多，2周最多，16周后仍维持较高水平；4周后骨骼肌的BrdU阳性细胞核数逐渐增加。移植后2周骨骼肌检测到少量人特异性抗核抗体（anti-Hu）阳性细胞核，4周后在骨骼肌肌膜可见少量人特异性Dys阳性细胞，8周后有Dy8mRNA的表达；随着移植时间的延长，anti-Hu抗体阳性细胞核数和Dys阳性细胞比例逐渐增高，Dys mRNA的表达逐渐增强。结论hBM-MSCs移植mdx鼠后早期（2—4周）主要分布在骨髓、肝、肺，后期（4周后）主要分布在骨髓、骨骼肌。Objective To investigate the dynamic distribution of human bone marrow mesenchymal stem cells （hBM-MSCs） in mdx mice. Methods Twenty-four 8-10-week-old immunocompromised mdx mice were transplanted with 1 ×10^7 passage 5 hBM-MSCs labeled with bromodeoxyuridine （BrdU） by means of injection into the tail vein. The mice were euthanized 48 hours and 2, 4, 8, 12, 16, 20, and 24 weeks after transplantation. BrdU-positive cells in tissue and organs of the mice were detected by immunofluorescence ana- lysis. Skeletal muscle was stained for anti-human nuclei mouse monoclonal antibody （anti-Hu） and analyzed for human dystrophin （Dys） expression by immunohistochemistry and reverse transcription-polymerase chain reaction. Results After transplantation, BrdU-positive cells were found in most organs （especially in bone marrow, liver, and lung） within 4 weeks, and these cells in liver and lung decreased gradually after 4 weeks. At 48 hours after transplantation, BrdU-positive cells were found in bone marrow, which reached a peak level after 2 weeks and were still detectable after 16 weeks. BrdU-positive cells in skeletal muscle increased gradually over time of transplantation. A small number of anti-Hu positive cells were detected in skeletal muscle 2 weeks after transplantation. A small number of Dys positive cell were seldom found at 4 weeks and small Dys mRNA expression detected 4 weeks after transplantation. The proportion of anti-Hu in parallel with Dys positive cells and Dys mRNA in skeletal muscle of mdx mice increased gradually over time of transplantation. Conclusion After being transplanted into mdx mice, hBM-MSCs are mainly distributed in bone marrow, liver, and lung during the early time （2-4 weeks）, and then in bone marrow and skeletal muscle （after 4 weeks）.

关 键 词：Duehenne型肌营养不良症 骨髓 间充质干细胞 分布 

分 类 号：R746.2[医药卫生—神经病学与精神病学]