J Nutr Health.  2018 Jun;51(3):208-214. 10.4163/jnh.2018.51.3.208.

Inhibitory effect of the aqueous extract of a tetraploid ‘etteum’ variety in Platycodon grandiflorum on degranulation and inflammatory mediator release in RBL-2H3 cells

Affiliations
  • 1Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University, Chuncheon, Gangwon 24252, Korea. myej4@hallym.ac.kr
  • 2Research Institute of Chamdahanbio, Chamdahanbio Ltd., Samcheok, Gangwon 25949, Korea.
  • 3Department of Food Science & Nutrition, Dongseo University, Busan 47011, Korea.
  • 4Department of Biochemistry, College of Medicine, Hallym University, Chuncheon, Gangwon 24252, Korea.

Abstract

PURPOSE
Platycodon grandiflorum (a domestic diploid variety, DV-PG) has been used as a food and component of various traditional oriental medicines. Although DV-PG is known to have an anti-allergic effect, little is known about the beneficial health effects of the tetraploid "˜Etteum' variety in the Platycodon grandiflorum (TV-PG), which is a recently developed variety. In this study, we investigated the effect of TV-PG on the rat basophilic leukemia mast cell (RBL-2H3)-mediated allergic response.
METHODS
To examine the effects of TV-PG on the allergic response, RBL-2H3 cells were sensitized with dinitropheny (DNP)-immunoglobin E, treated with various concentrations of TV-PG, and challenged with DNP-human serum albumin. We estimated cell granulation by measuring the release of β-hexosaminidase and production of inflammatory mediators by ELISA.
RESULTS
TV-PG had no effect on the proliferation or cytotoxicity of RBL-2H3 cells within the concentration range of 0 to 200 µg/mL. TV-PG inhibited degranulation of RBL-2H3 cells by antigen stimulation in a dose-dependent manner. TV-PG also suppressed the production of inflammatory cytokines and mediators such as interleukin-4, tumor necrosis factor-α, prostagladin E2, and leukotriene B4 in RBL-2H3 cells by antigen stimulation.
CONCLUSION
These results indicate that TV-PG exhibits anti-allergic activity via inhibition of degranulation as well as suppression of inflammatory mediators and cytokine release. These findings suggest that TV-PG may have potential as a preventive and therapeutic agent for the treatment of various allergic diseases.

Keyword

functional food; Platycodon grandiflorum; allergy; mast cell; inflammation mediators

MeSH Terms

Animals
Basophils
Cytokines
Diploidy
Enzyme-Linked Immunosorbent Assay
Functional Food
Hypersensitivity
Inflammation Mediators
Interleukin-4
Leukemia
Leukotriene B4
Mast Cells
Medicine, East Asian Traditional
Necrosis
Platycodon*
Rats
Serum Albumin
Tetraploidy*
Cytokines
Inflammation Mediators
Interleukin-4
Leukotriene B4
Serum Albumin

Figure

  • Fig. 1 Effect of TV-PG on the cell viability in RBL-2H3 cells. RBL-2H3 cells were plated at a density of 50,000 cells/well in 24-well plates with DMEM supplemented with 10% FBS. Twenty-four hours after plating, the cells were then incubated for 6 or 24 h in medium containing 0, 50, 100 or 200 µg/mL TV-PG or 200 g/mL DV-PG. Cell numbers were estimated by the MTT assay. Each bar represents the mean ± SEM (n = 6). Means at a time without a common letter differ, p<0.05. *p<0.05, **p<0.01, ***p<0.001 significantly different from that of 200 µg/mL TV-PG treated group. TV-PG = the aqueous extract of a tetraploid ‘Etteum’ variety in Platycodon grandiflorum, DV-PG = the aqueous extract of a domestic diploid variety in Platycodon grandiflorum.

  • Fig. 2 Effect of TV-PG on degranulation of β-hexosaminidase release from RBL-2H3 cells. RBL-2H3 cells were plated at a density of 2.5 × 105 cells/well in 24-well plates with DMEM supplemented with 10% FBS. Twenty-four hours after plating, the cells were incubated in a growth medium containing 50 ng/mL DNP-IgE (antibody) for 2 hours. After washed with siraganian buffer, incubated in siraganian buffer containing 5.6 mM CaCl2 and 0.1% BSA for 10 min, and then treated with 0, 50, 100 or 200 µg/mL TV-PG or 200 µg/mL DV-PG for 30 min. To activate the cells and evoke an allergic reaction, the cells were stimulated for 2 h with 10 µg/mL DNP-HSA. β-hexosaminidase secretion into the supernatant was measured and the inhibition of β-hexosaminidase (%) were estimated. Each bar represents the mean ± SEM (n = 6). Means without a common letter differ, p<0.05. *p<0.05, **p<0.01, *** p<0.001 significantly different from that of 200 µg/mL TV-PG treated group. TV-PG = the aqueous extract of a tetraploid ‘Etteum’ variety in Platycodon grandiflorum, DV-PG = the aqueous extract of a domestic diploid variety in Platycodon grandiflorum.

  • Fig. 3 Effects of TV-PG on the production of IL-4 (A) and TNF-α (B) in RBL-2H3 cells. The cells were sensitized with anti-DNP-IgE (50 ng/ml), stimulated by DNP-HSA (10 µg/ml) and treated various concentrations of TV-PG as described in Fig 2. Conditioned media were collected after 6 h. The concentrations of IL-6 and TNF-α in the conditioned media were measured using the relevant ELISA kits. Each bar represents mean ± SEM (n = 4). Means without a common letter differ, p < 0.05. * p < 0.05, ** p < 0.01, *** p < 0.001 significantly different from that of 200 µg/mL TV-PG treated group. TV-PG = the aqueous extract of a tetraploid ‘Etteum’ variety in Platycodon grandiflorum, DV-PG=the aqueous extract of a domestic diploid variety in Platycodon grandiflorum.

  • Fig. 4 Effects of TV-PG on the production of PGE2 (A) and LTB4 (B) in RBL-2H3 cells. The cells were sensitized with anti-DNP IgE (50 ng/ml), stimulated by DNP-HSA (10 µg/ml) and treated various concentrations of TV-PG as described in Fig 2. Conditioned media were collected after 6 h. The concentrations of PGE2 and LTB4 in the conditioned media were measured using the relevant ELISA kits. Each bar represents mean ± SEM (n = 4). Means without a common letter differ, p < 0.05. *p<0.05, **p<0.01, *** p < 0.001 significantly different from that of 200 µg/mL TV-PG treated group. TV-PG = the aqueous extract of a tetraploid ‘Etteum’ variety in Platycodon grandiflorum, DV-PG = the aqueous extract of a domestic diploid variety in Platycodon grandiflorum.


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