粒细胞集落刺激因子对急性辐射损伤小鼠胸腺细胞亚群及其近期输出功能的影响  

Effect of granulocyte colony-stimulating factor on murine thymic emigration and subsets reconstitution after a sublethal dose of irradiation

作　　者：赵红霞 郭梅[2] 孙雪冬[2] 艾辉胜[2] 

机构地区：[1]解放军第二炮兵总医院血液科, 北京,100088 [2]军事医学科学院附属医院

出　　处：《中华放射医学与防护杂志》2011年第6期-,共6页Chinese Journal of Radiological Medicine and Protection

摘　　要：目的 观察粒细胞集落刺激因子(G-CSF)对急性辐射损伤小鼠胸腺细胞亚群重建及近期输出功能的影响.方法 雌性BALB/c小鼠50只给于6 Gy 60Co 1次性全身照射后随机均分为GCSF组和对照组.GCSF组小鼠给予重组人G-CSF 100 μg·kg-1·d-1皮下注射,连续14 d,对照组小鼠给予等体积磷酸盐缓冲液(PBS)皮下注射,连续14 d.照后7、14、21和28 d颈部脱臼处死小鼠,取出胸腺制成单个核细胞悬液,使用流式细胞仪检测胸腺双阴性细胞发育4阶段(DN1 ～4)细胞比例的变化,以及CD4+ CD8+、CD4+ CD8 -、CD4 - CD8+细胞亚群的比例.使用荧光定量PCR的方法检测照后30和60 d 105个胸腺细胞中T细胞受体重排删除环(sjTREC)拷贝数,判断胸腺输出功能.结果 照后7d,胸腺DN细胞大量增殖,DN1细胞比例下降,DN4细胞比例增高,GCSF组DN1细胞比例在此时间点明显高于对照组(t =9.59,P＜0.05).照后21 d GCSF组DN3、DN4细胞比例均分别高于对照组(t =16.37、7.6,P＜0.05).照后7 dCD4+CD8+细胞比例降至最低,14 d出现反弹,21 d再次下降,以后逐渐恢复,照后28 d GCSF组CD4+ CD8+细胞比例恢复正常并明显高于对照组(t =12.22,P＜0.05).G-CSF对胸腺CD4+ CD8-细胞比例影响不大.照后21 d GCSF组CD4-CD8+细胞比例明显高于对照组(t=3.77,P＜0.05).荧光定量PCR结果提示照后30 d GCSF组胸腺细胞sjTREC拷贝数明显高于对照组(t=5.95,P＜0.01),但照后60 d两组胸腺细胞sjTREC拷贝数差异无统计学意义.结论 G-CSF可促进急性辐射损伤小鼠胸腺双阴性及双阳性细胞的增殖、分化,提高胸腺输出功能,加快中枢免疫重建.Objective To investigate the effects of recombinant human granulocyte colonystimulating factor(G-CSF) on murine thymic emigration and subsets reconstitution after a sublethal dose of irradiaton.Methods Female BALB/c mice were irradiated with a 6.0 Gy of γ-ray total-body irradiation and then randomly divided into GCSF group and control group.For mice in the GCSF group,recombinant human G-CSF 100 μg · kg-1 · d-1 was injected subcutaneously once daily for 14 continuous days and mice in the control group were given the same volume of phosphate buffered solution (PBS).At 7,14,21 and 28 days later,mice were killed and thymus mononuclear cell suspension were analyzed by flow cytometry for the percentage of the four stages of thymic CD4 -CD8 - double negative cells (DN1-4) and the CD4 + CD8 + double positive ( CD4 + CD8 + DP),CD4 + CD8 - single positive ( CD4 + SP),CD4 -CD8 + single positive cells (CD8 + SP).Real-time PCR was used for detection and quantitation of murine T cell receptor rearrangement excision circles(sjTRECs) of the thymic cells of 30 and 60 d after irradiation.Results The percentage of thymic DN1 cells in GCSF group was significantly higher than that of the control group 7 d after irradiation (t =9.59,P ＜ 0.05 ).21 d later,the proportion of thymic DN3 and DN4 cells were higher than those of the control group ( t =16.37,7.6,P ＜ 0.05 ).The percentage of thymic CD4 + CD8 + DP cells decreased 7 d after irradiation,increased at 14 d,decreased again at 21 days,and then got a permanent recover.The percentage of thymic CD4 + CD8 + DP cells in the GCSF group recovered to normal and was significantly higher than that of the control group 28 days after irradiation (t =12.22,P＜ 0.05).The percentage of thymic CD8 + SP cells of the GCSF group was significantly higher than that of the control group 21 d after irradiation ( t =3.77,P ＜ 0.05 ),while G-CSF had no obvious influence on the percentage of the thymic CD4 + SP cells.The sjTRECs copies in the GCSF group was significantly higher than that of the cont

关 键 词：粒细胞集落刺激因子 辐射损伤 免疫重建 

分 类 号：R285.5[医药卫生—中药学]