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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(Sca-1+)HSC/HPC,连续移植3代构建HSC/HPC衰老体内模型。60Coγ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Sca-1+HSC/HPC出现细胞衰老特征,随移植代数的增加,Sca-1+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Sca-1+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Sca-1+HSC/HPC衰老的作用。
Objective To investigate the effect of ginsenoside Rg1 on senescence-stimulating effect on the signal transduction axis of sirtuin 6 / nuclear factor-κB (SIRT6 / NF-κB) in hematopoietic stem / progenitor cells (HSC / HPC) Methods HSC / HPC was isolated and purified from male donor mouse MSCs by immunomagnetic sorting. HSC / HPC aging models were constructed by transplanting for 3 generations. 60Coγ-ray lethal dose irradiated female rats were divided into 4 groups, irradiation control group; aging model group; Rg1 treatment of aging group; Rg1 prevention of aging group. The effect of Rg1 on the regulation of Sca-1 + HSC / HPC senescence in vivo was analyzed by CFU-Mix culture, cell cycle analysis and senescence related β-galactosidase (SA-β-Gal) staining. Real-time quantitative PCR and Western blotting were used to detect the expression of SIRT6 and NF-κB mRNA and protein. Results Sca-1 + HSC / HPC showed the characteristics of cell senescence after transplantation. With the increase of transplantation algebra, the percentage of cells in the G0 / G1 phase of Sca-1 + HSC / G0 / G1 and the positive rate of SA- -Mix number drops. Compared with the aging model group, the percentage of cells in G0 / G1 phase of Sca-1 + HSC / HPC and the positive rate of SA-β-Gal in Rg1-treated aging group and Rg1-preventive aging group decreased, while CFU- High; SIRT6 mRNA and protein expression was up-regulated, NF-κB mRNA and protein expression was down; Rg1 prevention of aging indicators of each group than the Rg1 treatment of aging group was significantly. Conclusion Rg1 may play an important role in senescence of Sca-1 + HSC / HPC by regulating SIRT6 / NF-κB signaling axis.