重组人白介素-12对辐射损伤脐带血造血干细胞凋亡及其胞内STAT5、STAT4和丝裂原活化蛋白激酶表达的影响  

Effect of recombinant human interleukin-12 on apoptosis of hematopoietic stem cells isolated from umbilical cord blood after irradiation and intracellular expression of STAT5,STAT4 and mitogen activated protein kinase

作　　者：郑凤娇[1] 王翠玲 王增松 郭桂铃 李文棠 谢飞群 牛丽红 罗群方 张宜俊 

机构地区：[1]解放军第458医院检验科,广州510602 [2]广州市恺泰生物科技有限公司,广州510620

出　　处：《中国生物制品学杂志》2012年第9期1152-1155,共4页Chinese Journal of Biologicals

基　　金：科技部科技型中小企业技术创新基金(10C26214414767)

摘　　要：目的探讨重组人白介素-12(Recombinant human interleukin-12,rhIL-12)对辐射损伤新生儿脐带血造血干细胞凋亡及其胞内STAT5、STAT4和丝裂原活化蛋白激酶(Mitogen activated protein kinase,MAPK)表达的影响。方法收集16份新生儿脐带血,分离单个核细胞,经终浓度分别为0.01、0.1和1 ng/ml的rhIL-12处理24 h后,用直线加速器以4 Gy X射线单次照射24 h,用流式细胞仪分析CD34+造血干细胞的凋亡率;并观察CD34+造血干细胞经1 ng/ml rhIL-12体外活化15和30 min后,细胞内STAT5、STAT4和MAPK蛋白的表达水平。结果 rhIL-12对辐射损伤引起的新生儿脐带血CD34+造血干细胞凋亡具有明显的下调作用(P<0.01),且呈剂量依赖性(P<0.01);rhIL-12与新生儿脐带血CD34+造血干细胞共同孵育15和30 min,均可显著上调造血干细胞内STAT5和MAPK蛋白的表达水平(P<0.01或<0.001),下调STAT4蛋白的表达水平(P<0.05或<0.01),且均呈时间依赖性(P<0.001或<0.05)。结论 rhIL-12对辐射损伤的新生儿脐带血造血干细胞具有抗凋亡作用,并能上调造血干细胞内STAT5和MAPK蛋白的表达水平。Objective To investigate the effect of recombinant human interleukin-12（rhIL-12） on apoptosis of hematopoietic stem cells isolated from umbilical cord blood after irradiation as well as the íntracellular expressions of STAT5,STAT4 and mitogen activated protein kinase（MAPK）.Methods Mononuclear cells were isolated from 16 umbilical cord blood samples,and treated with rhIL-12 at final concentrations of 0.01,0.1 and 1 ng / ml for 24 h respectively,then irradiated by linear accelerator with X-ray at a dosage of 4 Gy for 24 h.The apoptosis rate of CD34＋ hematopoietic stem cells was determined by flow cytometry.The expressions of STAT5,STAT4 and MAPK in CD34＋ ematopoietic stem cells after activation with 1 ng / ml rhIL-12 for 15 and 30 min in vitro were observed.Results Recombinant human IL-12 down-regulated the apoptosis of CD3^4＋ hematopoietic stem cells isolated from umbilical cord blood after irradiation significantly（P〈0.01）,in a dose-dependent mode（P〈 0.01）.The expression levels of STAT5 and MAPK in CD3^4＋ hematopoietic stem cells after co-incubation with rhIL-12 for 15 and 30 min increased significantly（P 0.01 or P 〈0.001）,while that of STAT4 decreased significantly（P〈 0.05 or P〈 0.01）,in time-dependent modes.Conclusion The rhIL-12 showed anti-apoptotic effect on CD34＋ hematopoietic stem cells isolated from umbilical cord blood after irradiation,and increased the expression levels of STAT5 and MAPK in the cells.

关 键 词：白细胞介素-12 造血干细胞 辐射损伤 细胞凋亡 STAT5 STAT4 丝裂原活化蛋白激酶 

分 类 号：Q256[生物学—细胞生物学]