CD34^＋脐血干细胞转染pIRES2-FL—IL-3后静脉移植对辐射损伤小鼠的影响  

Effect of CD34^＋ cord blood stem cell transfected by plasmid vector pIRES2-FL-IL-3 on the mice after irradiation

作　　者：张勇[1] 张林生[1] 郭朝华[2] 张红宾[1] 董世武[2] 

机构地区：[1]第三军医大学西南医院血液科,重庆400038 [2]第三军医大学全军复合伤研究所

出　　处：《中华放射医学与防护杂志》2008年第3期228-231,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金青年基金资助项目（30200071）

摘　　要：目的观察CD34^＋脐血干细胞转染质粒载体pIRES2-FL-IL-3后静脉移植对辐射损伤小鼠的影响并探讨其机制。方法质粒载体pIRES2-FL-IL-3转染脐血CD34^＋干细胞，将其通过静脉移植到辐射损伤的小鼠体内（5×10^5个细胞，只，双表达组，12只），观察辐照后2、4和6周动物存活率及外周血象变化。6周时免疫荧光法检测实验小鼠脾脏CD34表达，RT-PCR法和Westernblot法进行IL-3和FL的mRNA和蛋白活性检测，另外3组为单纯CD34^＋细胞组（CD34组）、转染pIRES2-IL-3CD34^＋细胞组（IL-3组）和转染pIRES2-FL CD34^＋细胞组（FL组），各组动物数均为12只。结果CD34组、IL-3组和FL组动物6周存活率分别为25．0％、50．0％和50．0％，双表达组动物存活率为91．7％，显著高于其余各组。2周时双表达组白细胞数量、红细胞数量及血小板数量显著高于其余3组。双表达组脾脏CD34免疫荧光强度显著高于其余3组，IL-3及FL的mRNA水平及蛋白活性均显著高于其余3组。结论真核共表达质粒载体pIRES2-FL-IL-3转染CD34^＋脐血干细胞后对辐射损伤小鼠具有显著的促脾脏造血作用，其机制与移植后小鼠脾脏组织中移植CD34^＋脐血干细胞的聚集、增殖及目的蛋白的高效表达有关。Objective To observe the effect of CD34^＋ cord blood stem cell transfected by plasmid vector pIRES2-FL-IL-3 on the mouse after irradiation and to investigate its mechanisms. Methods In the co-expressed group （12 mice）, CD34^＋ cord blood stem cells were transfected by plasmid vector pIRES2-FL-IL-3. 5 × 10^5 cells were injected intravenously in the mouse. The hemogram changes in mice were detected 2, 4 and 6 weeks after radiation, At 6 weeks after irradiation, the expression of the CD34 in spleen was detected by immumofluorescence method. The mRNA level and the activity of IL-3 and FL were detected by RT-PCR and Western blot. Other 3 groups were CD34^＋ cell group （CD34 group）, pIRES2-IL-3 group（IL 3 group） and pIRES2-FL group（FL group）, and there were 12 mice in each group. Results The survival rate of CD34 group, IL3 group and FL group at the 6th week were 25.0% （3/12）, 50.0% （6/12） and 50.0% （6/12）, respectively. It was91.7%（11/12） in the co-expressed group, which was higher than those in the other groups. The expression of the CD34 of spleen in the co-expressed group was higher than those of the other groups. The mRNA level and the activity of IL-3 and FL of spleen in the co-expressed group were higher than those in the other groups too. Conclusions The CD34^＋ cord blood stem cells transfected by plasmid vector pIRES2-FL-IL-3 have hemogenesis promotion effect on the mice after irradiation, which was related with the aggregation, proliferation of stem cells and the high expression of the interest proteins.

关 键 词：白介素-3 FLT3配体 干细胞 辐射 造血 

分 类 号：R144[医药卫生—公共卫生与预防医学]