解剖学报

解剖学报 ›› 2015, Vol. 46 ›› Issue (5): 623-628.doi: 10.16098/j.issn.0529-1356.2015.05.008

• 细胞和分子生物学 • 上一篇    下一篇

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1* 王亚平2 王建伟2 丁继超1   

  1. 1. 云南省大理学院组织学胚胎学教研室, 云南 大理 671000; 2. 重庆医科大学干细胞与组织工程学研究室, 重庆 400016
  • 收稿日期:2015-02-02 修回日期:2015-03-27 出版日期:2015-10-06 发布日期:2015-10-06
  • 通讯作者: 周玥 E-mail:zhouyue_120@sina.com
  • 基金资助:

    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

Effects of sirtuin 6/nuclear factorκB signaling pathway in delaying hematopoietic stem cell and progenitor cell senescence with ginsenoside R1 during serial transplantation

ZHOU Yue 1* WANG Ya-ping2 WANG Jian-wei2 DING Ji-chao1   

  1. 1. Department of Histology and Embryology, Dali University, Yunnan Dali 671000, China; 2.Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, Chongqing 400016, China
  • Received:2015-02-02 Revised:2015-03-27 Online:2015-10-06 Published:2015-10-06
  • Contact: ZHOU Yue E-mail:zhouyue_120@sina.com

摘要：

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。

关键词:

Abstract:

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 modelin 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.

Key words: