人参皂苷Rg_1在造血干/祖细胞连续移植中对抗细胞衰老的作用研究

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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。方法:免疫磁性分选法分离纯化的雄性供体小鼠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:对照组、衰老模型组、Rg1治疗衰老组、Rg1延缓衰老组。荧光定量PCR检测受体鼠骨髓细胞Y染色体(Sry基因)表达,确定受体鼠重建造血细胞来源;观察受体鼠存活时间及外周血血象指标的恢复,确定受体鼠造血功能重建情况及Rg1促进造血恢复情况;造血祖细胞混合性集落(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Sca-1+HSC/HPC衰老的生物学特点及Rg1体内调控Sca-1+HSC/HPC衰老的作用。结果:雌性受体鼠重建造血细胞来源于雄性供体鼠;连续移植后受体鼠30 d存活率明显降低,外周血象恢复延缓,Sca-1+HSC/HPC出现细胞衰老特征: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延缓衰老组受体鼠30 d存活率,WBC,HCT,PLT增高,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。Rg1延缓衰老组变化较Rg1治疗衰老组明显。结论:人参皂苷Rg1在HSC/HPC连续移植过程中具有延缓和治疗Sca-1+HSC/HPC衰老的作用。Rg1延缓衰老作用优于Rg1治疗衰老作用。 Objective: To investigate the effect of ginsenoside Rg1 on anti-cell senescence in continuous transplantation of hematopoietic stem / progenitor cells (HSC / HPC). Methods: HSC / HPC aging model was established by three consecutive generations of Sca-1 + HSC / HPC isolated from mouse donor by immunomagnetic separation. 60Co γ-ray lethal dose irradiated female recipient rats were divided into 4 groups: control group, aging model group, Rg1 treatment of aging group, Rg1 anti-aging group. Fluorescent quantitative PCR was used to detect the expression of Y chromosome (Sry gene) in recipient mouse bone marrow cells to determine the source of reconstituted hematopoietic cells in recipient mice. The survival time of recipient mice and the recovery of peripheral blood hemoglobin indices were observed to determine the reconstruction of Rg1 Promote the hematopoietic recovery; culture of hematopoietic progenitor cells in mixed colonies (CFU-Mix), cell cycle analysis and aging-related β-galactosidase (SA-β-Gal) staining Sca-1 + HSC / HPC aging biology Features and Rg1 in vivo regulation of Sca-1 + HSC / HPC aging. Results: The reconstructed hematopoietic cells of female recipient mice were derived from male donors. After 30 days of continuous transplantation, the survival rate of recipients decreased significantly and the recovery of peripheral blood was delayed. The characteristics of cell senescence in Sca-1 + HSC / HPC were G0 / G1 phase The proportion of cells and the positive rate of SA-β-Gal staining increased, and the number of CFU-Mix decreased. Compared with the same aging model group, the survival rate of Rg1 treated group and Rg1 delayed aging group at 30 d, WBC, HCT, PLT increased, the proportion of cells in Sca-1 + HSC / HPC G0 / G1 phase, -Gal staining positive rate decreased CFU-Mix increased. Rg1 delayed aging group than the Rg1 treatment of aging group. CONCLUSION: Ginsenoside Rg1 can delay and treat Sca-1 + HSC / HPC senescence in HSC / HPC continuous transplantation. Rg1 anti-aging effect than Rg1 treatment of aging.
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