J Vet Sci.  2007 Mar;8(1):39-44. 10.4142/jvs.2007.8.1.39.

Radioprotective effects of an acidic polysaccharide of Panax ginseng on bone marrow cells

• Kim HJ
• Kim MH
• Byon YY
• Park JW
• Jee Y
• Joo HG

• KMID: 1104234
• DOI: http://doi.org/10.4142/jvs.2007.8.1.39

Abstract

An acidic polysaccharide of Panax ginseng (APG), so called ginsan is known to have important immunomodulatory activities. It was recently reported that APG has radioprotective effects in mice but the detailed mechanism was not fully elucidated. This study examined the effects of APG on bone marrow cells (BMs). The phenotypical and functional changes in APG-treated BMs after gamma radiation were studied. The benefit of APG on BMs damaged by gamma radiation was determined by measuring the cell viability. Using 2 different assays, a pretreatment with APG significantly increased the viability of BMs against gamma radiation. APG-treated BMs had a significantly higher amount of IL-12, which is a major cytokine for immune responses, compared with the medium-treated BMs. The expression of MHC class II molecules of APG-treated BMs was also increased, and APG-treated BMs showed significantly higher levels of allogeneic CD4+ T lymphocyte proliferation. Furthermore, APG-treated mice had a larger number of BMs after gamma radiation than the control mice, and the BMs of APG-treated mice were successfully cultured into dendritic cells, which are the representative antigenpresenting cells. Overall, this study shows that APG alters the phenotype of BMs, increases the viability and alloreactivity of BMs after gamma radiation both in vitro and in vivo. Therefore, APG may be a good candidate radioprotective agent for BMs.

Keyword

bone marrow cells; gamma radiation; ginsan; Panax ginseng; radioprotection

MeSH Terms

Animals
Bone Marrow Cells/*drug effects/radiation effects
CD4-Positive T-Lymphocytes/metabolism
Cell Survival/radiation effects
Flow Cytometry
Gamma Rays
Interleukin-12/biosynthesis
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Nitric Oxide/biosynthesis
Panax/*chemistry
Polysaccharides/*pharmacology
Radiation-Protective Agents/*pharmacology

Figure

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  Fig. 1 Acidic polysaccharide of Panax ginseng (APG) increased the viability of BMs. The cells were seeded in a 24-well culture plate and treated with 10 µg/ml of APG. After irradiation, trypan blue exclusion test was performed (A). The cells were seeded in a 96-well culture plate and treated with 0-10 µg/ml of APG. After irradiation, MTT assay was performed (B). All values are represented as mean ± SD and indicated statistical significance *p < 0.05 , ***p < 0.001, respectively.

• 

  Fig. 2 APG markedly increased IL-12 production in BMs but not nitric oxide (NO). The cells were seeded and treated with APG as described in Fig. 1A. After gamma irradiation, the cells were incubated for 24 h and the supernatants were harvested. The same supernatants were used in the IL-12 (A) and NO (B) assays. All values are represented as mean ± SD and indicated statistical significance **p < 0.01, ***p < 0.001, respectively.

• 

  Fig. 3 The enhanced expression of MHC class II molecules on the surface of APG-treated BMs. BMs were seeded and treated with APG as described in Fig. 1A. The cells were stained as described in M&M. The cells acquired by flow cytometric analysis were gated (A) and a representative histogram of each sample is shown (B). The number indicates the mean fluorescence intensity, minus that of the isotype control. The mean ± SD was obtained from 3 samples (C). All values are represented as mean ± SD and indicated statistical significance ***p < 0.001.

• 

  Fig. 4 APG increased the proliferation of CD4+ T lymphocytes. CD4+ T lymphocytes of Balb/c mice were co-cultured with BMs of C57BL/6 mice. After culturing for 5 days, the proliferation of CD4+ T lymphocytes was measured using 3H-thymidine incorporation assay. APG-treated BMs significantly increased the proliferation of allogeneic CD4+ T lymphocytes. The result is representative of 2 experiments. All values are represented as mean ± SD and indicated statistical significance **p < 0.01, ***p < 0.001.

• 

  Fig. 5 APG significantly increased the number of BMs and DC progenitor cells in vivo. The mice were injected with PBS alone or APG, and then irradiated. After 4 days, the number of BMs obtained from PBS-injected (control) or APG-injected mice was counted (A) and the cells were cultured in the presence of GM-CSF to verify the presence of DC progenitor cells. The number of the dot plot indicates the percentage of CD11c and MHC II double positive cells (B). Result is a representative of 2 individual experiments. All values are represented as mean ± SD and indicated statistical significance ***p < 0.001.

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