Journal List > J Nutr Health > v.51(3) > 1097793

J Nutr Health. 2018 Jun;51(3):208-214. Korean.
Published online June 28, 2018.  https://doi.org/10.4163/jnh.2018.51.3.208
© 2018 The Korean Nutrition Society
Inhibitory effect of the aqueous extract of a tetraploid ‘etteum’ variety in Platycodon grandiflorum on degranulation and inflammatory mediator release in RBL-2H3 cells
Jae In Jung,1 Hyung Seo Kim,2 Han Kyeol Ji,2 Hyun Sook Lee,3 Jae-Yong Lee,1,4 and Eun Ji Kim1
1Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University, Chuncheon, Gangwon 24252, Korea.
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.

To whom correspondence should be addressed. tel: +82-33-248-3106, Email: myej4@hallym.ac.kr
Received February 27, 2018; Revised April 01, 2018; Accepted April 16, 2018.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.


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.

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

Figures


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.
Click for larger image


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.
Click for larger image


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.
Click for larger image


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.
Click for larger image

Notes

This research was supported by the Ministry of Trade, Industry and Energy (MOTIE) and Korea Institute for Advancement of Technology (KIAT) through the Research and Promoting Regional Specialized Industry (R0005131) and by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (R1A6A3A01075809).

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