Lab Anim Res.  2017 Mar;33(1):32-39. 10.5625/lar.2017.33.1.32.

Immunomodulatory effects of ethanol extract of germinated ice plant (Mesembryanthemum crystallinum)

• Choi JH
• Jo SG
• Jung SK
• Park WT
• Kim KY
• Park YW
• Park JH

• KMID: 2375270
• DOI: http://doi.org/10.5625/lar.2017.33.1.32

Abstract

The purpose of this study was to investigate the immunomodulatory activity of ice plant (Mesembryanthemum crystallinum) extract (IPE) in vitro and in vivo. Raji (a human B cell line) and Jurkat (a human T cell line) cells were treated with various doses of IPE and cell proliferation was measured by WST assay. Results showed that IPE promoted the proliferation of both Raji and Jurkat cells in a dose-dependent manner. IPE also enhanced IL-6 and TNF-α production in macrophages in the presence of lipopolysaccharide (LPS), although IPE alone did not induce cytokine production. Moreover, IPE treatment upregulated iNOS gene expression in macrophages in a time- and dose-dependent manner and led to the production of nitric oxide in macrophages in the presence of IFNγ. In vivo studies revealed that oral administration of IPE for 2 weeks increased the differentiation of CD4+, CD8+, and CD19+ cells in splenocytes. These findings suggested that IPE has immunomodulatory effects and could be developed as an immunomodulatory supplement.

Keyword

Ice plant extract (IPE); lymphocyte; macrophage; cytokines; cell proliferation

MeSH Terms

Administration, Oral
Cell Proliferation
Cytokines
Ethanol*
Gene Expression
Humans
Ice*
In Vitro Techniques
Interleukin-6
Jurkat Cells
Lymphocytes
Macrophages
Mesembryanthemum*
Nitric Oxide
Cytokines
Ethanol
Ice
Interleukin-6
Nitric Oxide

Figure

• 

  Figure 1 Proliferative effect of ice plant extract treatment on B and T cells. Different concentrations of IPE treatment were applied to Raji (A) or Jurkat cells (B) for 24 h. Cell proliferation was determined as described in Materials and Methods. Data are presented as mean±SD (n=3). **P<0.01 vs control, ***P<0.001 vs control.

• 

  Figure 2 Effect of ice plant extract on proliferation and cytokine production in macrophages. Cells were treated with LPS (10 ng/mL) for 24 h in the absence or presence of the indicated dose of IPE. IL-6 (A) and TNF-α (B) levels in culture supernatants were measured by ELISA. Data are presented as mean±SD (n=3). ***P<0.001 vs LPS-treated group.

• 

  Figure 3 Effect of ice plant extract on the expression of iNOS mRNA in IFN-γ-stimulated macrophages. (A) Cells were treated with IPE (5 µL/mL) and co-treated with IFN-γ (100 ng/mL) for the indicated time. (B) Cells were treated with the indicated concentration of IPE and co-treated with IFN-γ (100 ng/mL) for 6 h. iNOS gene expression was analyzed using real-time qPCR and results were normalized to the expression of GAPDH. (C) Cells were treated with various concentrations of IPE for 24 h. The nitrite content in the medium was determined by Griess reagent. Data are shown as the mean±SD of triplicate samples from one representative experiment of three independent experiments. **P< 0.01 vs control, ***P< 0.001 vs control.

• 

  Figure 4 Population changes in lymphocyte subsets in mice administered ice plant extract for 2 weeks. On the day after the final administration, mice were sacrificed and their spleens were collected. Splenic single cells (1×105 cells/well) were stained with FITC- and PE-conjugated monoclonal antibodies specific for the mouse T cell markers CD4 (A), CD8 (B), and the mouse B cell marker CD19 (C). Stained cells were analyzed by flow cytometry using a BD Accuri C6. Values are reported as the percentage of each lymphocyte subset population expressing the CD marker.

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