摘要
背景:骨髓间充质干细胞在体外抑制再生障碍性贫血T淋巴细胞的活化与增殖,并有支持骨髓造血作用。目的:观察骨髓间充质干细胞对再生障碍性贫血模型鼠存活率的影响,探讨其治疗再生障碍性贫血的机制。设计、时间及地点:随机对照动物实验,于2009-01/04在安徽医科大学第一附属医院中心实验室完成。材料:清洁级雌性BALB/C小鼠40只,8~10周龄,体质量16~18g,作为受体。清洁级DBA/2小鼠5只,6~10周龄,体质量15~19g为淋巴细胞供体。方法:取BALB/c小鼠骨髓进行间充质干细胞体外增殖培养,传至3~5代的细胞用于实验。建立免疫介导再生障碍性贫血小鼠模型。40只BALB/C小鼠随机分为3组,治疗组(n=15):造模后将1×106个骨髓间充质干细胞输注到治疗组小鼠体内进行治疗;模型组(n=15):仅造模不进行任何治疗;对照组(n=10):不进行任何干预。主要观察指标:分别于第1,2,3,4周观察各组小鼠的存活率;流式细胞仪及酶联免疫吸附法测定骨髓间充质干细胞治疗后第28天各组小鼠T淋巴细胞亚群及血浆肿瘤坏死因子α的表达。结果:①模型组小鼠3只于第2周内死亡,7只于第3周内死亡,2只于第4周内死亡,28天存活率20%;治疗组小鼠4只于第3周内死亡,28天存活率73%。②与模型组相比,治疗组和对照组CD4的表达增高[(27.12±4.23)%,(32.18±6.73)%,(38.44±7.88)%,P<0.01];CD8和肿瘤坏死因子α表达均降低[CD8:(34.42±5.13)%,(27.33±5.09)%,(23.31±5.24)%,P<0.01;肿瘤坏死因子α:(47.27±10.11),(12.34±5.87),(9.31±3.77)ng/L,P<0.01]。结论:骨髓间充质干细胞可以提高再生障碍性贫血小鼠存活率,其治疗机制可能与调节T淋巴细胞亚群及抑制T淋巴细胞分泌的肿瘤坏死因子α有关。
BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) inhibit activation and proliferation of T cells in vitro and support hematopoiesis of bone marrow.
OBJECTIVE: To observe the effect of BMSCs on survival rate of mice with aplastic anemia and to explore the mechanism for treating aplastic anemia.
DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Central Laboratory of the First Affiliated Hospital of Anhui Medical University from January to April 2009.
MATERIALS: A total of 40 female BALB/c mice, clean grade, 8 10 weeks old, weighing 16 18 g, were used as recipients; another 5 DBA/2 mice, clean grade, 6-10 weeks old, weighing 15 19 g, were used as donors.
METHODS: BMSCs were extracted from BALB/c mice. The third-passage cells were used to establish aplastic anemia injury. A total of 40 BALB/c mice were randomly divided into three groups: treatment (1×10^6 BMSCs, n=15), model (modeling alone, n=15), and control (without any intervention, n=10) groups. MAIN OUTCOME MEASURES: Survival rate was observed at 1, 2, 3, and 4 weeks after operation; T lymphocyte subgroup and tumor necrosis factor e (TNF-o) expression were measured at day 28 postoperatively using flow cytometry and enzyme linked immunosorbent assay.
RESULT: (1) In the model group, three mice died at 2 weeks, seven at 3 weeks, and two at 4 weeks, and the survival rate was 20% at day 28 postoperatively. In the treatment group, four mice died at 3 weeks, and the survival rate was 73% at day 28 postoperatively. (2) As compared with the model group, CD4 expression increased in both treatment and control groups [(27.12±4.23)%, (32.18±6.73)%, (38.44±7.88)%, P 〈 0.01]; CD8 and TNF-α expression decreased [CDS: (34.42±5.13)%, (27.33±5.09)%, (23.31±5.24)%, P 〈 0.01; TNF-e: (47.27±10.11 ), (12.34±5.87), (9.31±3.77) ng/L, P 〈 0.01].
CONCLUSION: BMSCs enhance survival rate of mice with aplastic anemia, and the mechanism may be related to regulating T lymphocyte subgroup and inhibiting secretion of TNF-α.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第36期7138-7142,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
安徽省淮北市科协基金资助项目(080243)"骨髓间充质干细胞治疗再生障碍性贫血的实验研究"~~