欢迎访问《解剖学报》官方网站！今天是
English
人参皂苷Rg1在造血干/祖细胞连续移植中对延缓细胞衰老的作用与去乙酰化酶6/核因子-κB信号轴的关系
周玥* 王亚平 王建伟 丁继超
解剖学报 ›› 2015, Vol. 46 ›› Issue (5) : 623-628.
人参皂苷Rg1在造血干/祖细胞连续移植中对延缓细胞衰老的作用与去乙酰化酶6/核因子-κB信号轴的关系
Effects of sirtuin 6/nuclear factorκB signaling pathway in delaying hematopoietic stem cell and progenitor cell senescence with ginsenoside R1 during serial transplantation
目的 探讨人参皂苷Rg1在造血干/祖细胞（HSC/HPC）连续移植中对抗细胞衰老的作用与去乙酰化酶6/核因子-κB(SIRT6/NF-κB）信号轴的关系。 方法 免疫磁性分选法分离纯化雄性供体小鼠干细胞抗原阳性（Sca1+）HSC/HPC，连续移植3代构建HSC/HPC衰老体内模型。60Co γ射线致死剂量辐射雌性受体鼠后分4组，照射对照组；衰老模型组；Rg1治疗衰老组；Rg1预防衰老组。造血祖细胞混合集落（CFU-Mix）培养，细胞周期分析和衰老相关β-半乳糖苷酶（SA-β-Gal）染色分析Rg1体内调控Sca-1+HSC/HPC衰老的作用。实时定量PCR及Western blotting检测衰老调控分子SIRT6、NF-κB mRNA及蛋白的表达。 结果 连续移植后受体鼠Sca1+HSC/HPC出现细胞衰老特征，随移植代数的增加，Sca1+HSC/HPC G0/G1期细胞比例及SA-β-Gal染色阳性率增高，CFU-Mix数量下降。与同代衰老模型组相比，Rg1治疗衰老组及Rg1预防衰老组受体鼠Sca1+ HSC/HPC G0/G1期细胞比例、SA-β-Gal染色阳性率下降，CFU-Mix数量升高；SIRT6 mRNA及蛋白表达上调，NF-κB mRNA及蛋白表达下调；Rg1预防衰老组各指标变化均较Rg1治疗衰老组明显。结论 Rg1可能通过调控SIRT6/NF-κB信号轴发挥其对抗连续移植过程中Sca1+ HSC/HPC衰老的作用。
Objective To investigate the effect of sirtuin(SIRT)6/nuclear factor(NF)-κB signaling pathway in delaying hematopoietic stem cell (HSC) and progenitor cell(HPC) senescence with ginsenoside Rg1 during serial transplantation. Methods Sca-1+HSC/HPC from male donator mouse was isolated and purified by magnetic activated cell sorting (MACS). Sca-1+HSC/HPC replicability aged modelin vivo was established through the Sca-1+HSC/HPC serial transplantation. The femal recipient mice radiated lethal dose from 60Co γ ray were divided into four groups: the control group, the aged model group, the Rg1 treat aged group and the Rg1 prevent aged group. The effect of Rg1 to delay Sca-1+HSC/HPC senencence in vivo was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining, mixed hematopoietic progenitor cell culture and cell cycle assay. The expressions of SIRT6 and NF-κB mRNA and protein were detected by quantitative PCR and Western blotting. Results With the increasing of transplantation, Sca-1+HSC/HPC appeared aging characters. The number of cells entered G0/G1 phase and the percentage of SA-β-gal positive cells was increased, and the number of CFU-Mix was decreased. Compared with the aged model group, the number of cells entered G0/G1 phase and the percentage of SA-β-gal positive cells were decreased, the number of CFU-Mix was increased to Sca-1+ HSC/HPC in Rg1 treat aged group and Rg1 prevent aged group, the expression of SIRT6 mRNA and protein was up regulated and the expression of NF-κB mRNA and protein was down regulated in Rg1 treat aged group and Rg1 prevent aged group.The changes of Rg1 prevent aged group was significantly higher than Rg1 treat aged group. Conclusion Rg1 could resist Sca-1+HSC/HPC senescence during serial transplantation in which the signaling pathway of SIRT6-NF-κB may play an important role.
人参皂苷Rg1 /
去乙酰化酶6 /
核因子-κB /
干细胞抗原阳性造血干/祖细胞 /
衰老 /
实时定量PCR /
免疫印迹法 /
小鼠
Ginsenoside Rg1 /
Sirtuin6 /
Nuclear factor-κB /
Sca-1+ HSC/HPC /
Senescence /
Real\ /
time PCR /
Western blotting /
Mouse
［1］Chu SF, Zhang JT. New achievements in ginseng resrarch and its future prospects［J］. Chin J Integr Med, 2009, 15(6):403-408.
［2］Yang L, Zhang J, Zheng K, et al. Long-term ginsenoside Rg1 supplementation improves age-related cognitive decline by promoting synaptic plasticity associated protein expression in C57BL/6J mice［J］. J Gerontol A Biol Sci Med Sci, 2014, 69 (3): 282-294.
［3］Wang YP, eds. Stem Cell Senescence and Disease［M］. Beijing: Science Press, 2009:1-22. (in Chinese)
王亚平,主编.干细胞衰老与疾病［M］. 北京：科学出版社，2009：1-22.
［4］Tennen RI, Berber E, Chua KF.Functional dissection of SIRT6: identification of domains that regulate histone deacetylase activity and chromatin localization［J］. Mech Ageing Dev, 2010, 131(3):185-192.
［5］Zhou Y, Yang B, Yao X, et al. Establishment of an aging model of Sca-1+hematopoietic stem cell and studies on its relative biological mechanisms［J］. In Vitro Cell Dev Biol Anim, 2011, 47(2): 149-153.
［6］Farese AM，Hunt P，Grab LB，et al. Combined administration of recombinant human megakaryocyte growth and development factor and gran ulocyte colony-stimulating factor enhances multilineage hematopoietic reconstitution in nonhuman primates after radiation-induced malTow aplasia［J］. J Clin Invest, 1996, 97（9)：2145-2151.
［7］Zhang XP, Zhang GH, Wen KM, et al. Oxidation low density lipoprotein promotes aging of mouse hematopoietic stem cells through regulating cell cycles［J］. Acta Anatomica Sinica, 2013, 44(2):189-193. (in Chinese)
张先平，张贵海，文坤明,等. 氧化低密度脂蛋白可促进小鼠造血干细胞衰老［J］. 解剖学报，2013，44(2)：189-193.
［8］Chen X, Zhang J, Fang Y, et al. Ginsenoside Rg1 delays tert-butyl hydroperoxide-induced premature senescence in human WI-38 diploid fibroblast cells［J］. J Gerontol A Biol Sci Med Sci, 2008, 63(3):253-264.
［9］Shen LH, Zhang JT. Ginsenoside Rg1 promotes proliferation of hippocampal progenitor cells［J］. Neurol Res, 2004, 26(4):422-428.
［10］Zhou Y, Yang B, Jiang R, et al. Mechanism of ginsenoside Rg1 to delay hematopoietic stem cell senescence［J］. National Medical Journal of China, 2010, 90（48）：3421-3425. (in Chinese)
周玥，杨斌，姜蓉，等.人参皂苷Rg1延缓造血干细胞衰老及其相关机制［J］.中华医学杂志，2010，90（48）：3421-3425.
［11］TenNapel MJ, Lynch CF, Burns TL, et al. SIRT6 minor allele genotype is associated with >5-year decrease in lifespan in an aged cohort［J］. PLoS One, 2014, 9 (12): 1-18.
［12］Lee N, Kim DK, Kim ES, et al. Comparative interactomes of SIRT6 and SIRT7: Implication of functional links to aging［J］. Proteomics, 2014, 14 (13): 1610-1622.
［13］Kawahara TL, Michishita E, Adler AS, et al. SIRT6 links histone H3 lysine 9 deacetylation to NF-kappaB-dependent gene expression and organismal life span［J］. Cell, 2009, 136(1):62-74.
［14］Ashburner BP, Westerheide SD, Baldwin AS Jr. The p65 (RelA) subunit of NF-kappaB interacts with the histone deacetylase (HDAC) corepressors HDAC1 and HDAC2 to negatively regulate gene expression［J］.Mol Cell Biol, 2001, 21(20):7065-7077.
衰老调控因子SIRT6与NF-κB对话在人参皂苷Rg1延缓造血干/祖细胞衰老中的作用;当归多糖调控血液干细胞衰老的机理研究
/
| 〈 |
|
〉 |
