骨髓间充质干细胞输注对重型再生障碍性贫血治

　　【摘要】 本研究探讨人源骨髓间充质干细胞（MSC）输注对免疫介导性再生障碍性贫血模型小鼠骨髓造血功能及生存率的影响.采用雌性BALB/c小鼠30只，参照姚军等方法建立免疫介导性再生障碍性贫血模型，实验分模型组、MSC组及照射组。MSC组于制模后3天从小鼠尾静脉1次性输注人骨髓MSC 1×106，观察各组小鼠外周血象变化、第28天存活率、骨髓有核细胞数量、造血集落生成和骨髓病理学特征。结果显示：于制模后第7天，模型组外周血WBC 显著低于MSC组和照射组，分别为（0.65±0.05）×109/L，（2.45±0.71）×109/L，（2.30±0.33）×109/L；第10天3组均表现为全血细胞减少，第14天模型组血象指标仍持续降低，而MSC组及照射组血象指标开始回升，至28天3组小鼠存活率分别为18.2%、80%和100%，MSC组小鼠存活率显著高于模型组（p<0.01）。MSC组小鼠单侧股骨有核细胞数量、造血祖细胞集落生成数量均显著高于模型组，而与照射组结果一致。结论：MSC输注可减轻再生障碍性贫血模型小鼠骨髓造血衰竭程度，提高存活率。

　　Effect of Bone Marrow Mesenchymal Stem Cell Infusion on Hemato poiesis in Mice with Aplastic Anemia

　　Abstract To explore the effects of infusion with human mesenchymal stem cells from bone marrow on bone marrow hematopoietic function and survival in mice with aplastic anemia (AA), immuno瞞ediated aplastic anemia mice model was established according to Yao′s method. Thirty BALB/c female mice were divided into AA model group, MSC group and radiation group. In MSC group, 1×106 MSC was infused by intravenous injection at 3 days after establishment of model. Thechanges of blood count， the number of nuclear cells in a single thigh瞓one, colony瞗orming units(CFU), pathological features of bone marrow and survival rate of mice were observed in all groups. The results showed that the number of white blood cells in peripheral blood of AA model group at day 7 after establishment of model was （0.65±0.05）×109/L , whichwas significantly lower than those in MSC group and radiation alone group, respectively. Pancytopenia was found in all three groups at day 10. At day 14 the pancytopenia was continued in AA model group while it was less severe in MSC and radiation groups. In MSC group the number of nucleated cells in single thigh bone and pathological slice as well as CFU睪M were significantly higher than those in AA model group, respectively. The survival rate of mice in MSC group was 80.0% which was higher than that of group in AA model (18.2%, p<0.01).There were no significant differences between MSC group and radiation alone group in bone marrow hematopoiesis and survival rate. It is concluded that MSC infusion reduces the degree of bone marrow failure and improve survival of immuno瞞ediated AA mice model.

　　KEY WORDS APLASTIC ANEMIA； ANIMAL MODEL； MESENCHYMAL STEM CELL； HEMATOPOIESIS

　　骨髓间充质干细胞（MESENCHYMAL STEM CELLS, MSC）除具有促进造血干细胞移植后造血重建外，还具有免疫调节作用，防治移植物抗宿主病［1，2］。获得性再生障碍性贫血主要是T淋巴细胞介导的骨髓造血功能衰竭［3，4］，且再生障碍性贫血患儿骨髓MSC体外增殖能力较正常对照儿童明显减弱［5］。本研究探讨人源骨髓MSC 对免疫介导性再生障碍性贫血小鼠骨髓造血功能及生存率的影响，再生障碍性贫血模型的建立VU`B5'&Ai^yP e, ZBy3egqgf^R3~]www.６６wｅｎ．ｃｏｍ0\~^A o5eHZ( kEthW_d*2ch+&D1

　　清洁级雌性BALB/C小鼠（H2D、MLSB），8-10周龄，16-18 G，作为受者。清洁级DBA/2小鼠（H2D、MLSA），6-10周龄，为淋巴细胞供者。两者均由中山大学动物实验中心提供。参照文献［6］建立免疫介导性再生障碍性贫血模型。取DBA/2小鼠颈椎脱臼处死，常规消毒，无菌取出胸腺及颈部、颌下、腋窝、腹股沟、肠系膜等处淋巴结，分别获胸腺、淋巴结细胞悬液,细胞浓度为5×106/ML，按胸腺细胞与淋巴结细胞以1∶2比例混合，即胸腺淋巴结细胞悬液。台盼蓝鉴定细胞活性，活性细胞>95%。雌性BALB/C小鼠经5 GY 60CO Γ采湎呷身照射（剂量率125.35 CGY/MIN），照射后20小时内由尾静脉注