骨髓干细胞输注修复放疗免疫抑制的动物实验

Influence of Bone Barrow Stem Cell Transplantation on Immunosuppressed Mice

目的探讨骨髓干细胞(bone marrowstem cell,BMSC)对放疗所致免疫抑制小鼠的修复作用,为干细胞用于临床修复放化疗后免疫抑制的患者提供一定的理论依据.方法选择雌性BALB/C小鼠,随机分为实验组、对照组和空白组,实验组和对照组的每只小鼠经照射剂量为8 Gy的6 MV直线加速器X线照射,随后实验组小鼠通过尾静脉输注小鼠骨髓干细胞1×107个,0.2 m L,对照组小鼠通过尾静脉输入0.2 m L生理盐水,空白组小鼠不做任何处理.于移植后第15、25、35天用血细胞分析仪检测外周血血常规,流式细胞仪检测T淋巴细胞亚群(CD3+、CD4+、CD8+)、ELISA试剂盒检测血清免疫球蛋白Ig M变化情况,并观察生存状态和存活率.结果经放疗所致免疫抑制的小鼠经骨髓干细胞输注后35 d存活率为80%,未经输注的小鼠35 d存活率为10%;放疗后小鼠经骨髓干细胞输注后外周血白细胞数、淋巴细胞比例及CD3+T细胞亚群较未经输注干细胞的小鼠高(P<0.05);在骨髓干细胞移植后第15天,CD4+%、CD8+%和免疫球蛋白Ig M最低,随后逐渐升高(P<0.05);骨髓干细胞移植后能降低免疫抑制小鼠外周血单核细胞比例及CD4+/CD8+比值(P<0.05).结论骨髓干细胞的输注能够修复免疫抑制小鼠的免疫功能,这为干细胞用于临床修复放化疗所致的免疫抑制的病人提供了一定的实验基础和理论依据.

Objective To investigate the influence of bone marrow stem cell transplantation onimmunosuppressed mice induced by radiotherapy.Methods Female BALB/C mice were randomly divided intothree groups.The mice in test group and in control group were irradiated with X-ray at dosage of8Gy respectively,while those in blank group were not irradiated.After that,bone marrow stem cells1x107in0.2ml normal salinewere injected into mice by tail veins in test group,while0.2ml normal saline(NS)was injected into mice by tailvein in control group.The peripheral blood was subjected to routine blood test on days15,25,35aftertransplantation,T lymphocytes(CD3+,CD4+,CD8+)were determined by flow cytometry(FCM)on days15,25,35after transplantation,and the concentration of mice peripheral blood immunoglobulins IgM weremeasured by the ELISA method.All the mice were observed for survivals for35days.Results The survival rates ofmice35d after Hematopoietic stemcell transplantation were80%,while10%in control group.The leukocyte countsand Lymph%as well as CD3+%in peripheral blood after transplantation increased as compared with those receivedNS(P<0.05).However,CD4+%,CD8+%and serum immunoglobulin IgM after transplantation decreased,which decreased remarkably on day15and begun to recover after that day.BMSC decreased Mono%and CD4+/CD8+in peripheral blood(P<0.05).Conclusion Bone marrow stem cell transplantation can repair the suppressedimmunity of mice.