摘要
本研究探讨粒细胞集落刺激因子(G-CSF)对急性辐射损伤小鼠胸腺细胞的细胞周期、凋亡及近期输出功能的影响。雌性BALB/c小鼠40只,给予6 Gy 60Coγ线1次性全身照射后随机均分为2组。G-CSF组小鼠给予重组人G-CSF 100μg/(kg.d),皮下注射,连续14天;对照组小鼠给予等体积磷酸盐缓冲液,皮下注射,连续14天。用流式细胞仪检测照后72小时内胸腺细胞的细胞周期及凋亡细胞比例;应用荧光定量PCR方法检测照后30和60天105个胸腺细胞中T细胞受体重排删除环(sjTREC)拷贝数,以判断胸腺细胞输出功能。结果表明,G-CSF在照后6小时即可使胸腺细胞出现G0/G1期阻滞,G-CSF组vs对照组为(82.0±5.0)%vs(75.9±2.8)%(p<0.05),S期细胞比例下降,G-CSF组vs对照组为(10.2±4.8)%vs(15.7±2.3)%(p<0.05),但对G2/M期细胞比例影响不大。胸腺细胞受照后6小时即出现明显凋亡,12小时达高峰。G-CSF组在照后6及12小时胸腺细胞凋亡率分别为(11.5±2.4)%和(15.5±3.3)%,而对照组分别为(16.5±2.2)%和(22.6±0.7)%,两时间点统计学差异明显(p<0.05)。荧光定量PCR结果显示,照后30天G-CSF组胸腺细胞sjTREC拷贝数明显高于对照组(p<0.01),但照后60天两组胸腺细胞sjTREC拷贝数无统计学差异。结论:G-CSF可调节急性辐射损伤后胸腺细胞周期,减少胸腺细胞凋亡,增加胸腺细胞输出功能,促进中枢免疫恢复。
This study was aimed to investigate the effect of recombinant human granulocyte colony-stimulating factor（G-CSF） on murine thymocyte emigration and cell cycle alteration after a sublethal dose of gamma-irradiation.Female BALB/c mice were given 6.0 Gy γ-ray total body irradiation and then randomly divided into G-CSF and control groups.Mice in the G-CSF group were injected recombinant human G-CSF 100 μg/（kg· d） subcutaneously once daily for 14 consecutive days and mice in the control group were given the same volume of phosphate buffered solution.Thymocyte cycle alteration and the proportion of apoptosis cells were detected by flow cytometry within 72 hours after irradiation.Real-time PCR was used for detection and quantitation of murine T cell receptor rearrangement excision circles（sjTREC）of the thymic cells at 30 and 60 day after the irradiation.The results showed that at 6 hour after irradiation G-CSF could significantly increase the thymic cells in G0/G1 phase,G-CSF vs control：（82.0±5.0）% vs（75.9±2.8）%（p0.05）,and decrease the thymic cells in S phase,G-CSF vs control：（10.2±4.8）% vs（15.7±2.3）%（p0.05）,but G-CSF seemed have no evident effects on the percentage of thymic cells in G2/M phase.G-CSF could also protect thymocytes from apoptosis at 6 hour and 12 hour after irradiation the percentages of apoptosis cells in G-CSF group were（11.5±2.4）% and（15.5±3.3）%,respectively,which were significantly lower than that of the control group（16.5±2.2）% and（22.6±0.7）%,respectively（p0.05）.The sjTREC copy amount was conspicuously higher in G-CSF group than that in the control at 30 day after irradiation（p0.01）,but the preponderance disappeared 60 days later.It is concluded that G-CSF has a positive effect on the thymic cell cycle alteration to protect thymocytes from apoptosis and enhance the recent thymocyte emigrantion,which may contribute to the central immune reconstitution after irradiation.
出处
《中国实验血液学杂志》
CAS
CSCD
2011年第5期1294-1298,共5页
Journal of Experimental Hematology
关键词
粒细胞集落刺激因子
辐射
胸腺细胞周期
胸腺输出功能
granulocyte colony-stimulating factor
irradiation
thymic cell cycle
thymic emigration
