Nutr Res Pract.  2012 Apr;6(2):97-105.

Suppressive effects of Schizandra chinensis Baillon water extract on allergy-related cytokine generation and degranulation in IgE-antigen complex-stimulated RBL-2H3 cells

• Chung MJ
• Kim JM
• Lee S
• Kim T
• Kim D
• Baek J
• Kim T
• Lee J
• Kim K
• Yoon JA
• Choe M

Abstract

Schizandra chinensis Baillon is a traditional folk medicine plant that is used to treat and prevent several inflammatory diseases and cancer in Korea, but the underlying mechanisms involved in its anti-allergic activity are not fully understood. This study was designed to investigate mechanisms of anti-allergic activity of a Schizandra chinensis Baillon water extract (SCWE) in immunoglobulin E (IgE)-antigen complex-stimulated RBL2H3 cells and to assess whether gastric and intestinal digestion affects the anti-allergic properties of SCWE. Oxidative stress is an important consequence of the allergic inflammatory response. The antioxidant activities of SCWE increased in a concentration-dependent manner. RBL-2H3 cells were sensitized with monoclonal anti-dinitrophenol (DNP) specific IgE, treated with SCWE, and challenged with the antigen DNP-human serum albumin. SCWE inhibited beta-hexosaminidase release and expression of interleukin (IL)-4, IL-13, and tumor necrosis factor-alpha (TNF-alpha) mRNA and protein in IgE-antigen complex-stimulated RBL2H3 cells. We found that digested SCWE fully maintained its antioxidant activity and anti-allergic activity against the IgE-antigen complex-induced activation of RBL-2H3 cells. SCWE may be useful for preventing allergic diseases, such as asthma. Thus, SCWE could be used as a natural functional ingredient for allergic diseases in the food and/or pharmaceutical industries.

Keyword

Allergic diseases; antioxidant; cytokine; RBL-2H3 cells; Schizandra chinensis Baillon

MeSH Terms

Asthma
beta-N-Acetylhexosaminidases
Digestion
Drug Industry
Immunoglobulin E
Immunoglobulins
Interleukin-13
Interleukins
Korea
Medicine, Traditional
Oxidative Stress
Plants
RNA, Messenger
Schisandra
Serum Albumin
Tumor Necrosis Factor-alpha
Water
Immunoglobulin E
Immunoglobulins
Interleukin-13
Interleukins
RNA, Messenger
Serum Albumin
Tumor Necrosis Factor-alpha
Water
beta-N-Acetylhexosaminidases

Figure

• 

  Figure 1 RBL-2H3 cell toxicity to the Schisandra chinensis water extract (SCWE) and digested SCWE (DSCWE) and their antioxidant effects in a cell-free system. A and B, Cytotoxicity. After a 24 h treatment with various SCWE or DSCWE concentrations (0, 100, 250, 500, and 1,000 µg/mL), RBL-2H3 cell viability was determined by the MTT assay. C and D, Total phenol content. The Folin-Ciocalteu method was used. E, DPPH radical scavenging activity. Various concentrations of SCWE or DSCWE (0, 100, 250, 500, and 1,000 µg/mL) were mixed with the DPPH solution and incubated at 37℃, and the absorbance change at 517 nm was then determined. The reference compound was ascorbic acid. Means with different letters (a-e) differ significantly from each other (P < 0.05), as determined by Duncan's multiple range test (n = 4). Cell viability results were not significantly different (P > 0.05).

• 

  Figure 2 Effect of Schisandra chinensis water extract (SCWE) and digested SCWE (DSCWE) on β-hexosaminidase release in RBL-2H3 cells stimulated by the anti-DNP IgE-antigen (DNP-HSA) complex. RBL-2H3 cells (2 × 105 cells) in 24-well plates were preincubated with 0.5 µg/mL anti-DNP IgE for 12 h, washed with Siraganian buffer, incubated in Siraganian buffer containing 5.6 mM CaCl2 and 0.1% BSA for 10 min, and then treated with 1 mL of SCWE or DSCWE (0, 100, 250, 500, and 1,000 µg/mL) for 30 min. Cells were stimulated for 2 h with DNP-HSA (10 µg/mL) to activate the cells and evoke an allergic reaction. β-hexosaminidase secretion into the supernatant was then measured. Means with different letters (a-f) differ significantly from each other (P < 0.05), as determined by Duncan's multiple range test (n = 4). control, C.

• 

  Figure 3 Effects of Schisandra chinensis water extract (SCWE) and digested SCWE (DSCWE) on the expression of interleukin (IL)-4, IL-13 and tumor necrosis factor (TNF)-α mRNA in RBL-2H3 cells stimulated with the IgE-antigen (DNP-HSA) complex. RBL-2H3 cells were preincubated with anti-DNP IgE (0.5 µg/mL) for 12 h, treated with various concentration (0, 100, 250, 500, and 1,000 µg/mL) of SCWE or DSCWE for 30 min, and stimulated with DNP-HSA (1 µg/mL) for 2 h. The cells were then washed, lysed, and subjected to reverse transcription-polymerase chain reaction (RT-PCR). The IL-4, IL-13, and TNF-α mRNA levels in each sample were normalized to GAPDH levels. mRNA band density was quantified using SigmaGel software, and the group data were averaged and plotted. Means for each gene with different letters (a-e) differ significantly from each other (P < 0.05), as determined by Duncan's multiple range test (n = 4). Control, C.

• 

  Figure 4 Effects of Schisandra chinensis water extract (SCWE) on interleukin (IL)-4 and IL-13 protein production in RBL-2H3 cells stimulated with the IgE-antigen (DNP-HSA) complex. The cells were primed with anti-DNP IgE (0.5 µg/mL) for 12 h, treated with various concentrations (0, 100, 250, 500, and 1,000 µg/mL) of SCWE for 30 min, stimulated with DNP-HSA (1 µg/mL) for 2 h, washed, lysed, and subjected to Western blot analysis. IL-4 and IL-13 protein levels in each sample were normalized to α-tubulin levels. Protein band density was quantified using SigmaGel software, and the group data were averaged and plotted. Means for each protein with different letters (a-f) differ significantly from each other (P < 0.05), as determined by Duncan's multiple range test (n = 4). Control, C.

• 

  Figure 5 Effects of digested Schisandra chinensis water extract (DSCWE) on interleukin (IL)-4 and IL-13 protein production in RBL-2H3 cells stimulated with the IgE-antigen (DNP-HSA) complex. The cells were primed with anti-DNP IgE (0.5 µg/mL) for 12 h, treated with various concentrations (0, 100, 250, 500, and 1,000 µg/mL) of DSCWE for 30 min, stimulated with DNP-HSA (1 µg/mL) for 2 h, washed, lysed, and subjected to Western blot analysis. IL-4 and IL-13 protein levels in each sample were normalized to α-tubulin levels. Protein band density was quantified using SigmaGel software, and the group data were averaged and plotted. Means for each protein with different letters (a-d) differ significantly from each other (P < 0.05), as determined by Duncan's multiple range test (n = 4). Control, C.

• 

  Figure 6 Effects of Schisandra chinensis water extract (SCWE) and digested SCWE (DSCWE) on the tumor necrosis factor (TNF)-α protein production in RBL-2H3 cells stimulated with the IgE-antigen (DNP-HSA) complex. RBL-2H3 cells were preincubated with anti-DNP IgE (0.5 µg/mL) for 12 h, treated with various concentrations (0, 100, 250, 500, and 1,000 µg/mL) of SCWE or DSCWE for 30 min, and stimulated with DNP-HSA (1 µg/mL) for 2 h. The supernatant was subjected to enzyme-linked immunoabsorbent assay (ELISA). Means with different letters (a-e) differ significantly from each other (P < 0.05), as determined by Duncan's multiple range test (n = 4). Control, C.

Reference

1. Itoh T, Ohguchi K, Iinuma M, Nozawa Y, Akao Y. Inhibitory effect of xanthones isolated from the pericarp of Garcinia mangostana L. on rat basophilic leukemia RBL-2H3 cell degranulation. Bioorg Med Chem. 2008; 16:4500–4508. PMID: 18328716.

2. Matsuda H, Wang Q, Matsuhira K, Nakamura S, Yuan D, Yoshikawa M. Inhibitory effects of thunberginols A and B isolated from Hydrangeae Dulcis Folium on mRNA expression of cytokines and on activation of activator protein-1 in RBL-2H3 cells. Phytomedicine. 2008; 15:177–184. PMID: 17950587.

3. Huang F, Yamaki K, Tong X, Fu L, Zhang R, Cai Y, Yanagisawa R, Inoue K, Takano H, Yoshino S. Inhibition of the antigen-induced activation of RBL-2H3 cells by sinomenine. Int Immunopharmacol. 2008; 8:502–507. PMID: 18279805.

4. Chang GT, Kang SK, Kim JH, Chung KH, Chang YC, Kim CH. Inhibitory effect of the Korean herbal medicine, Dae-Jo-Whan, on platelet-activating factor-induced platelet aggregation. J Ethnopharmacol. 2005; 102:430–439. PMID: 16125889.

5. Hancke JL, Burgos RA, Ahumada F. Schisandra chinensis (Turcz.) Baill. Fitoterapia. 1999; 70:451–471.

6. Greene LS. Asthma, oxidant stress, and diet. Nutrition. 1999; 15:899–907. PMID: 10575668.

7. Shon MY, Choi SD, Kahng GG, Nam SH, Sung NJ. Antimutagenic, antioxidant and free radical scavenging activity of ethyl acetate extracts from white, yellow and red onions. Food Chem Toxicol. 2004; 42:659–666. PMID: 15019191.

8. Ryu JS, Hyun JW, Kim HW, Shim MJ, Kim BK. Effects of artificial digestive juice on the antitumor-immunity activity of protein-bound polysaccharide from Ganoderma lucidum. Yakhak Hoeji. 2000; 44:347–353.

9. Folin O, Denis W. A colorimetric method for the determination of phenols (and phenol derivatives) in urine. J Biol Chem. 1915; 22:305–308.

10. Mărghitaş LA, Stanciu OG, Dezmirean DS, Bobiş O, Popescu O, Bogdanov S, Campos MG. In vitro antioxidant capacity of honeybee-collected pollen of selected floral origin harvested from Romania. Food Chem. 2009; 115:878–883.

11. Choi HJ, Chung MJ, Ham SS. Antiobese and hypocholesterolaemic effects of an Adenophora triphylla extract in HepG2 cells and high fat diet-induced obese mice. Food Chem. 2010; 119:437–444.

12. Yamashita K, Suzuki Y, Matsui T, Yoshimaru T, Yamaki M, Suzuki-Karasaki M, Hayakawa S, Shimizu K. Epigallocatechin gallate inhibits histamine release from rat basophilic leukemia (RBL-2H3) cells: role of tyrosine phosphorylation pathway. Biochem Biophys Res Commun. 2000; 274:603–608. PMID: 10924324.

13. Watanabe J, Shinmoto H, Tsushida T. Coumarin and flavone derivatives from estragon and thyme as inhibitors of chemical mediator release from RBL-2H3 Cells. Biosci Biotechnol Biochem. 2005; 69:1–6. PMID: 15665459.

14. Fukao T, Terauchi Y, Kadowaki T, Koyasu S. Role of phosphoinositide 3-kinase signaling in mast cells: new insights from knockout mouse studies. J Mol Med (Berl). 2003; 81:524–535. PMID: 12928787.

15. Scalbert A, Morand C, Manach C, Rémésy C. Absorption and metabolism of polyphenols in the gut and impact on health. Biomed Pharmacother. 2002; 56:276–282. PMID: 12224598.

16. Zhao Z, Egashira Y, Sanada H. Ferulic acid is quickly absorbed from rat stomach as the free form and then conjugated mainly in liver. J Nutr. 2004; 134:3083–3088. PMID: 15514279.

17. Lafay S, Gil-Izquierdo A, Manach C, Morand C, Besson C, Scalbert A. Chlorogenic acid is absorbed in its intact form in the stomach of rats. J Nutr. 2006; 136:1192–1197. PMID: 16614403.

18. Aura AM. Microbial metabolism of dietary phenolic compounds in the colon. Phytochem Rev. 2008; 7:407–429.

19. Nishikawa H, Kitani S. Tea catechins have dual effect on mast cell degranulation induced by compound 48/80. Int Immunopharmacol. 2008; 8:1207–1215. PMID: 18602066.

20. Ramalakshmi K, Jagan Mohan Rao L, Takano-Ishikawa Y, Goto M. Bioactivities of low-grade green coffee and spent coffee in different in vitro model systems. Food Chem. 2009; 115:79–85.

21. Wijngaard HH, Rössle C, Brunton N. A survey of Irish fruit and vegetable waste and by-products as a source of polyphenolic antioxidants. Food Chem. 2009; 116:202–207.

22. Lee SH, Bae EA, Park EK, Shin YW, Baek NI, Han EJ, Chung HG, Kim DH. Inhibitory effect of eupatilin and jaceosidin isolated from Artemisia princeps in IgE-induced hypersensitivity. Int Immunopharmacol. 2007; 7:1678–1684. PMID: 17996677.

23. Kühn R, Rajewsky K, Müller W. Generation and analysis of interleukin-4 deficient mice. Science. 1991; 254:707–710. PMID: 1948049.

24. Hines C. The diverse effects of mast cell mediators. Clin Rev Allergy Immunol. 2002; 22:149–160. PMID: 11975420.

25. Grünig G, Warnock M, Wakil AE, Venkayya R, Brombacher F, Rennick DM, Sheppard D, Mohrs M, Donaldson DD, Locksley RM, Corry DB. Requirement for IL-13 independently of IL-4 in experimental asthma. Science. 1998; 282:2261–2263. PMID: 9856950.

26. Yang G, Li L, Volk A, Emmell E, Petley T, Giles-Komar J, Rafferty P, Lakshminarayanan M, Griswold DE, Bugelski PJ, Das AM. Therapeutic dosing with anti-interleukin-13 monoclonal antibody inhibits asthma progression in mice. J Pharmacol Exp Ther. 2005; 313:8–15. PMID: 15644434.

27. Gordon JR, Galli SJ. Mast cells as a source of both preformed and immunologically inducible TNF-alpha/cachectin. Nature. 1990; 346:274–276. PMID: 2374592.

28. Quan Z, Lee YJ, Yang JH, Lu Y, Li Y, Lee YK, Jin M, Kim JY, Choi JH, Son JK, Chang HW. Ethanol extracts of Saururus chinensis suppress ovalbumin-sensitization airway inflammation. J Ethnopharmacol. 2010; 132:143–149. PMID: 20699114.