人参皂苷Rg_1延缓造血干细胞衰老与p16^(INK4a)表达关系的研究  

Experimental study of relationship between effect of ginsenoside Rg_1 to delay hematopoietic stem cell senescence and expression of p16^(INK4a)

作　　者：周玥[1] 杨斌[1] 姚欣[1] 王亚平[1] 

机构地区：[1]重庆医科大学干细胞与组织工程研究室,重庆400016

出　　处：《中国中药杂志》2011年第5期608-613,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30973818);重庆市科委自然科学基金重点项目(CSTC;2009BA5038)

摘　　要：目的:探讨人参皂苷单体Rg1延缓造血干细胞(HSC)衰老与p16INK4a的表达调控关系,为寻找延缓HSC衰老途径提供理论和实验依据。方法:免疫磁性分选法分离纯化Sca-1+HSC后分组。对照组常规培养;衰老组运用三丁基过氧化氢(t-BHP)复制衰老模型;Rg1组在对照组基础上加入10μmol.L-1 Rg1共培养;Rg1延缓衰老组给予10μmol.L-1 Rg1预处理后,复制衰老模型;Rg1治疗衰老组,衰老模型复制后,给予10μmol.L-1Rg1抗衰老处理。衰老相关β-半乳糖苷酶(SA-β-gal)细胞化学染色、流式细胞术分析细胞周期和造血祖细胞混合集落(CFU-Mix)培养确定Rg1延缓或治疗Sca-1+HSC衰老生物学作用。RT-PCR及Western blotting检测衰老相关基因p16INK4a mRNA及蛋白的表达。结果:Rg1延缓衰老组,Rg1治疗衰老组与衰老组相比,SA-β-gal染色阳性细胞百分比降低,G1期细胞比例下降,生成造血祖细胞混合集落数增加,p16INK4a mRNA及蛋白的表达下降;Rg1延缓衰老组的SA-β-gal染色阳性细胞百分比、G1期比例、p16INK4a mRNA及蛋白的表达均低于Rg1治疗衰老组,形成造血祖细胞混合集落数高于Rg1治疗衰老组。结论:Rg1具有延缓和治疗Sca-1+HSC衰老的作用,Rg1延缓衰老比治疗衰老效果更好。Rg1可能通过调控p16INK4a的表达发挥其对抗t-BHP诱导的Sca-1+HSC衰老的作用。Objective：To investigate the relation between the effect of ginsenoside Rg1 to delay hematopoietic stem cell senescence and the expression of p16INK4a.The purpose is to provide the theory and experimental foundation for searching the methods of how to delay HSC senescence.Method：Sca-1＋HSC was isolated by magnetic cell sorting（MACS） and divided into five groups.The control group cells were routinely cultured,the aging group cells were induced aging by tert-butylhydroperoxide（t-BHP,final concentration of 100 μmol·L-1） to establish the aging model,the Rg1 group cells were co-cutured with Rg1（final concentration is 10 μmol·L-1）.To Rg1 delay aging group,Sca-1＋HSC were established aging model after pretreatment of Rg1（final concentration is 10 μmol·L-1）.To Rg1 treat aging group,Sca-1＋HSC gave Rg1（final concentration is 10 μmol·L-1） antiaging treatment after the aging model was established.The changes of cells observed by senescence-associated β-galactosidase（SA-β-gal） staining,cell cycle analysis and culture of mixed hematopoietic progenitor cell were used to investigate the antiaging and delay aging effect of ginsenoside Rg1.The expression of senescence associated p16INK4a mRNA and p16INK4a protein was examined by RT-PCR and western blotting.Result：Compared with aging group,the percentage of positive cells expressed SA-β-gal and cells in G1 phase decreased and the number of forming colony of mixed hematopoietic progenitor increased and it showed higher expression of p16INK4a mRNA and p16INK4a protein in Rg1 treat aging group and Rg1 delay aging group.Furthermore the percentage of positive cells expressed SA-β-gal,cells in G1 phase,the number of forming colony of mixed hematopoietic progenitor and the expression of p16INK4a mRNA and protein decreased in Rg1 delay aging group compared with Rg1 treat aging group.Conclution：Rg1 can significantly delay and treat the senescence of Sca-1＋HSC.The effect of Rg1 delaying aging is better than treatment.p16INK4a may be play a ke

关 键 词：人参皂苷RG1 P16INK4A 造血干细胞 衰老 

分 类 号：R285[医药卫生—中药学]