Ease effect of ginsenoside on different-intensity ionizing radiation damage to human hematopoietic stem cells

doi:10.3969/j.issn.2095-4344.2015.01.022

Abstract

Abstract:

BACKGROUND: Many domestic and foreign scholars and institutions are studying how to relieve radiation damage and to find the most suitable drug, while ginsenosides as the main pharmacological ingredient of ginseng show significant antioxidant effect.
OBJECTIVE: To investigate the ease effect of ginsenosides on human hematopoietic stem cells under different intensities of ionizing radiations.
METHODS: The CD34⁺ hematopoietic stem cells were isolated from the healthy cord blood. Then the cells were divided into normal group and ginsenoside-pretreated group, respectively, exposed under 1, 2, 5 Gy of X-ray irradiations for 24 hours. Cell viability was detected in irradiated hematopoietic stem cells by MTT assay. Apoptosis was estimated using the following assays: Annexin-V assay, caspase-3 mRNA and protein levels. The generation of reactive oxygen species was evaluated, in the presence or absence of ginsenoside in liquid cultures of CD34⁺ human hematopoietic stem cells irradiated with 1-, 2- and 5-Gy X-rays, using a flow cytometry assay. The Nrf-2 mRNA and protein levels were also studied by western blot analysis and RT-PCR, respectively.
RESULTS AND CONCLUSION: Ionizing radiation at the therapeutic dose could decrease the viability of CD34⁺ cells and induce the cell apoptosis, and meanwhile, the activity of intracellular reactive oxygen species also showed a progressive increase that was correlated with the dose of ionizing radiation. However, ginsenoside pretreatment could relieve these above-mentioned effects. Ginsenoside inhibited the increase in caspase-3 activity induced by ionizing radiation, and additionally, enhanced the mRNA and protein expressions of Nrf-2 in CD34⁺ cells. In conclusion, ginsenoside protects CD34⁺ hematopoietic stem cells from radiation effects, which is probably correlated with anti-apoptosis and anti-oxidant roles of ginsenosides.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: Hematopoietic Stem Cells, Ginsenosides, Radiation, Ionizing

CLC Number:

R394.2

Huang Ying, Liang Xiao-yan, Li Cheng-jin, Hu Jun, Zhou Li-qian. Ease effect of ginsenoside on different-intensity ionizing radiation damage to human hematopoietic stem cells[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(1): 124-129.

Figures/Tables 3

2.1 电离辐射诱导造血干细胞活性下降，发生凋亡以及诱导大量活性氧释放首先对0，1，2，5 Gy辐射量下造血干细胞的细胞活性进行了测定，结果见图1A。结果显示，随着辐射剂量的增加，存活细胞百分比呈现出进行性下降的趋势，而且辐射组与未辐射组之间差异有显著性意义(P < 0.05)。选取AV/PI双染中AV+细胞作为凋亡细胞进行了计数，统计学分析见图1B，可见随着辐射剂量的增加，AV+的造血干细胞数显著增多，也表现为进行性的上升趋势。之后，分别选取辐射后30 min，24 h两个时间点，对各实验组细胞内活性氧的含量进行了测定，由图1C可以看到，造血干细胞在受到辐射30 min后即可出现不同程度的活性氧释放，其中2 Gy和5 Gy组的释放量与0 Gy组相比差异有显著性意义(P < 0.05)。而当造血干细胞经电离辐射诱导24 h后，细胞内活性氧释放量增多更明显，其中1 Gy组和2 Gy组与0 Gy组相比差异有显著性意义(P < 0.05)。还需要注意的是，5 Gy组在24 h时间点检测到的活性氧释放量与0 Gy组差异无显著性意义(P > 0.05)。 2.2 人参皂苷对电离辐射诱导造血干细胞出现的凋亡及活性氧释放有一定的缓解作用选用了5 μmol/L的人参皂苷对经受电离辐射的造血干细胞进行了预处理后，发现电离辐射诱导造血干细胞出现的凋亡及活性氧释放有一定的缓解作用。首先，图1A中MTT结果显示，人参皂苷预处理后，造血干细胞活性与未处理组相比明显增强(P < 0.05)；同时，AV/PI的结果(图1B)也表明，经人参皂苷预处理后AV+细胞百分比也明显减少(P < 0.05)；另外，活性氧检测结果显示(图1C，D)，5 μmol/L的人参皂苷预处理后，各辐射剂量组活性氧释放量差异均无显著性意义，但5.0 mol/L人参皂苷组与未处理组相比差异均有显著性意义(P < 0.05)。这也表明了人参皂苷在一定程度上可以抑制电离辐射诱导的造血干细胞活性的下降、凋亡和活性氧的释放。"

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