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쥐 비만세포 (RBL-2H3)와 OVA/alum 감작 마우스에서 치자 추출물의 알레르기 염증 억제 효과

The Effects of Gardenia jasminoides J.Ellis on Allergic Inflammation in RBL-2H3 Mast Cells and OVA/alum-sensitized Mice

  • 김경리 (대전대학교 한의과대학 소아과학교실) ;
  • 이진화 (대전대학교 한의과대학 소아과학교실) ;
  • 김은진 (대전대학교 한의과대학 소아과학교실) ;
  • 김윤희 (대전대학교 한의과대학 소아과학교실)
  • Kim, Kyeong Ri (Department of Pediatrics, College of Korean medicine, Dajeon University) ;
  • Lee, Jin Hwa (Department of Pediatrics, College of Korean medicine, Dajeon University) ;
  • Kim, Eun Jin (Department of Pediatrics, College of Korean medicine, Dajeon University) ;
  • Kim, Yun Hee (Department of Pediatrics, College of Korean medicine, Dajeon University)
  • 투고 : 2018.10.26
  • 심사 : 2018.11.19
  • 발행 : 2018.11.30

초록

Objectives The purpose of this study is to investigate the effects of Gardenia jasminoides for. grandiflora extracts' (GAJ) anti-inflammatory effect on RBL-2H3 mast cells and OVA/alum-sensitized mice. Methods In this study, IL-4 and IL-13 production was measured via ELISA analysis, and mRNA expressions of GM-CSF, IL-4, IL-5, $TNF-{\alpha}$, IL-6 were analyzed by real-time PCR. In addition, MAPKs and $NF-{\kappa}B$ p65 transcription factors were examined using western blotting, and ELISA was used to understand IgE, IL-4, and IL-13 production in ovalbumin-allergic mice in in vitro study. Results As a result of this study, 1. GAJ were observed to suppress the mRNA expression of GM-CSF, IL-4, IL-13, IL-5, $TNF-{\alpha}$, IL-6 in comparison to PMA 50 ng/ml, ionomycin $0.5{\mu}M$ (PI) control group. 2. GAJ also inhibited the IL-4, IL-13 production in comparison to PI control group. 3. Western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65. 4. Orally-administered GAJ group in OVA/alum induced Balb/c mice showed decreased level of OVA-specific IgE in the serum. This group also has shown decreased the level of IL-4, IL-13 in the splenocyte culture supernatant. Conclusions Obtained results suggest that GAJ may regulate the allergic inflammation by transcription factors MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65 causing inhibition of Th2 cytokines in mast cells and OVA/alum-sensitized mice.

키워드

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Fig. 1. Cytotoxic effects of GAJ on RBL-2H3 mast cells

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Fig. 2. Suppressive effects of GAJ on GM-CSF mRNA expression

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Fig. 3. Suppressive effects of GAJ on IL-4 mRNA expression

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Fig. 4. Suppressive effects of GAJ on IL-13 mRNA expression

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Fig. 5. Suppressive effects of GAJ on IL-5 mRNA expression

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Fig. 6. Suppressive effects of GAJ on TNF-a mRNA expression

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Fig. 7. Suppressive effects of GAJ on IL-6 mRNA expression

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Fig. 8. Suppressive effects of GAJ on IL-4 production

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Fig. 9. Suppressive effects of GAJ on IL-13 production

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Fig. 10. Effects of GAJ on ERK phosphorylation in PI-induced RBL-2H3 mast cells

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Fig. 11. Effects of GAJ on p38 phosphorylation in PI-induced RBL-2H3 mast cells

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Fig. 12. Effects of GAJ on JNK phosphorylation in PI-induced RBL-2H3 mast cells

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Fig. 13. Effects of GAJ on the nuclear translocation of NF-κB p65 in PI-induced RBL-2H3 mast cells

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Fig. 14. Effect of GAJ on OVA-specific IgE release in OVA challenged allergic mouse model

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Fig. 15. Effect of GAJ on IL-4 release in release in OVA challenged isolated splenocytes

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Fig. 16. Effect of GAJ on IL-13 release in release in OVA challenged isolated splenocytes

Table 1. Rat Primer Sequence for Real-Time PCR Analysis

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참고문헌

  1. Korean Medicine University Herbal Medicine Commission. Herbal medicine. Seoul: Yeongrim Publishing Co. 2008:208-9.
  2. Debnath T, Park PJ, Deb Nath NC, Samad NB, Park HW, Lim BO. Antioxidant activity of Gardenia jasminoides Ellis fruit extracts. Food Chem. 2011;128(3):697-703. https://doi.org/10.1016/j.foodchem.2011.03.090
  3. Lin YJ, Lai CC, Lai CH, Sue SC, Lin CW, Hung CH, Lin TH, Hsu WY, Huang SM, Hung YL, Tien N, Liu X, Chen CL, Tsai FJ. Inhibition of enterovirus 71 infections and viral IRES activity by Fructus gardeniae and geniposide. Eur J Med Chem. 2013;62:206-13. https://doi.org/10.1016/j.ejmech.2012.12.038
  4. Toriizuka K, Kamiki H, Ohmura NY, Fujii M, Hori Y, Fukumura M, Hirai Y, Isoda S, Nemoto Y, Ida Y. Anxiolytic effect of Gardeniae Fructus-extract containing active ingredient from Kamishoyosan (KSS), a Japanese traditional Kampo medicine. Life Sci. 2005; 77(24):3010-20. https://doi.org/10.1016/j.lfs.2004.12.054
  5. Min DH, Kim DK, Lim JP, Yang JH. Transdermal drug delivery & therapeutic effect of the preparations of Lithospermi Radix and Gardeniae Fructus extracts on the burn & wound healing. J Kor Pharm Sci. 2005;35(4):255-63.
  6. Yang JH, Lee NH. Skin permeation and anti-inflammatory effects of hydrolyzed products of Gardeniae Fructus extracts. J Kor Pharm Sci. 2004;34(2):115-23.
  7. An HS. Textbook of pediatrics. 9th ed. Seoul: Korea Textbook Publishing Co. 2010:1104-21.
  8. Kang KY, Kim YH, Han JK, The suppressive effect on Th2 cytokines expression and the signal transduction mechanism in MC/9 mast cells by PRAL. J Pediatr Korean Med. 2014;28(2):23-39. https://doi.org/10.7778/jpkm.2014.28.2.023
  9. Park Y. Moisturizers in atopic dermatitis. J Skin Barrier Res (Korean). 2007;9(1):37-40.
  10. Abbas AK, Lichtman AH, Pillai S. Cellular & molecular immunology. 6th ed. Seoul: Epublic Publishing Co. 2008:451-71.
  11. Kim JW. Atopic dermatitis at allergic and immunological aspects. J Korean Dermatol Assoc. 2003;41(6):687-9.
  12. Renz H, Jujo K, Bradley KL, Domenico J, Gelfand EW, Leung DY. Enhanced IL-4 production and IL-4 receptor expression in atopic dermatitis and their modulation by interferon-gamma. J Invest Dermatol. 1992; 99(4):403-8. https://doi.org/10.1111/1523-1747.ep12616114
  13. Cha JH, Lim EM. Effects of Gardeniae Fructus on cytokines in mouse macrophage. J Korean Obstet Gynecol. 2014; 27(1):1-16. https://doi.org/10.15204/JKOBGY2014.27.1.001
  14. Sea S. Research for the effect of Gardeniae Fructus on regulatory T cell stimulation. Master Dissertation, Kyung Hee University, 2010.
  15. Ishiyama M, Miyazono Y, Sasamoto K, Ohkura Y, Ueno K. A highly water-soluble disulfonated tetrazolium salt as a chromogenic indicator for NADH as well as cell viability. Talanta. 1997;44(7):1299-305. https://doi.org/10.1016/S0039-9140(97)00017-9
  16. Leung DY, Guttman-Yassky E. Deciphering the complexities of atopic dermatitis: shifting paradigms in treatment approaches. J Allergy Clin Immunol. 2014;134(4):769-79. https://doi.org/10.1016/j.jaci.2014.08.008
  17. Cork MJ, Danby SG, Vasilopoulos Y, Hadgraft J, Lane ME, Moustafa M, Guy RH, Macgowan AL, Tazi-Ahnini R, Ward SJ. Epidermal barrier dysfunction in atopic dermatitis. J Invest Dermatol. 2009;129(8):1892-908. https://doi.org/10.1038/jid.2009.133
  18. National Health Insurance Corporation. Environment day right environment -related diseases health insurance representative 3 payment data analysis {Internet}. Available from: http://www.datanews.co.kr/news/article.html?no=94723.
  19. Jang HS. Diagnosis and treatment of atopic dermatitis. J Korean Acad Fam Med. 2002;23(7):831-40.
  20. Korea Centers for Disease Control and Prevent, Hanrim University College of Medicine Seongsim Hospital Pediatrics. The use of alternative medicine and management in children with atopic dermatitis. Korean J Pediatr. 2008;18(2):148-57.
  21. Chin HW, Jang HS, Jang BS, Jo JH, Kim MB, Oh CK, Kwon KS, Kwon YW. A study on utilization of alternative medicine for patients with atopic dermatitis. Korean J Dermatol. 2005;43(7):903-11.
  22. Kim SH. The use of oriental and complementary alternative medicine in children with atopic dermatitis. Master Dissertation, University of Cha Medical University, 2007.
  23. Koh DJ, You HJ, Lee JY, Kim DG. Clinical assessment of efficacy, safety and usefulness of traditional herbal medicine (THM) on atopic dermatitis. J Korean Orient Pediatr. 2006;20(2);129-37.
  24. Kang TJ, Lee SR. Causes and treatment management of atopic dermatitis. BioWave. 2010;12(3):1-25.
  25. Izuhara K, Shirakawa T, Adra CN, Hamasaki N, Hopkin JM. Emerging therapeutic targets in allergic disease. Pharmacol Ther. 2005;25(3):381-9.
  26. Gibbs BF, Haas H, Wolff HH, Grabbe J. Early IgE-dependent release of IL-4 and IL-13 from leukocytes is restricted to basophils: a comparison with other granulocytes and mononuclear cells. Inflamm Res. 2000; 49(Suppl1):S9-10. https://doi.org/10.1007/PL00000197
  27. Valent P, Spanbloochl E, Sperr WR, Sillaber C, Zsebo KM, Agis H, Strobl H, Geissler K, Bettelheim P, Lechner K. Induction of differentiation of human mast cells from bone marrow and peripheral blood mononuclear cells by recombinant human stem cell factor/kit-ligand in long-term culture. Blood. 1992;80(9):2237-45.
  28. Kim HC. Korean medicine pharmacology. Seoul: Jibmun Publishing Co. 2001:125-7.
  29. Lee CB. Coloured flora of Korea. Seoul: Hyangmunsa. 2003.
  30. Alavizadeh SH, Hosseinzadeh H. Bioactivity assessment and toxicity of crocin: a comprehensive review. Food Chem Toxicol. 2014;64:65-80. https://doi.org/10.1016/j.fct.2013.11.016
  31. Shin YW, Kim DW, Kim NJ. Studies on the processing of crude drugs (VII) -on the constituents and biological activities of Gardeniae Fructus by processing-. Korean J Pharmacogn. 2003;34(1):45-54.
  32. Kim HJ, Kim EJ, Seo SH, Shin CG, Jin C, Lee YS. Vanillic acid glycoside and quinic acid derivatives from Gardeniae Fructus. J Nat Prod. 2006;69(4):600-3. https://doi.org/10.1021/np050447r
  33. Chun SC, Jee SY, Lee SK. The antimicrobial activity of Naesohwangryuntang and its composition oriental medicine. Kor J Herbology. 2004;19(4):51-60.
  34. Choi CS, Sin SS, Chung SI, Yang YT. Antibacterial activity of soluble extracts of Gardenia jasminoides against mycobacteria other than Tubercle Bacilli. J Korea Soc Microbiol. 1986;21(1):53-62.
  35. Kim MJ, Nam YO, Yang JH. Anti-inflammatory and sedative effect of rat by the gel containing ketorolac and hydrolysis products of Gardeniae Fructus. J Korean Acad Dent Health. 2004;28(4):464-72.
  36. Han YN, Oh HK, Hwang KH, Lee MS. Antioxidant components of Gardenia Fruit. Kor J Pharmacogn. 1994: 25(3):226-32.
  37. Hwang KH, Park TK. Inhibitory activity of the fruit extract of Gardenia jasminoides on Monoamine Oxidase. Kor J Pharmacogn. 2007;38(2):108-12.
  38. Koo HJ, Lim KH, Jung HJ, Park EH. Anti-inflammatory evaluation of gardenia extract, geniposide and genipin. J Ethnopharmacol. 2006;103(3):496-500. https://doi.org/10.1016/j.jep.2005.08.011
  39. Wang GF, Wu SY, Rao JJ, Lu L, Xu W, Pang JX, Liu ZQ, Wu SG, Zhang JJ. Genipin inhibits endothelial exocytosis via nitric oxide in cultured human umbilical vein endothelial cells. Acta Pharmacol Sin. 2009;30(5):589-96. https://doi.org/10.1038/aps.2009.31
  40. Prieschl EE, Gouileux-Gruart V, Walker C, Harrer NE, Baumruker T. A nuclear factor of activated T cell-like transcription factor in mast cells is involved in IL-5 gene regulation after IgE plus antigen stimulation. J Immunol. 1995;154(11):6112-9.
  41. Aversa G, Punnonen J, Cocks BG, de Waal Malefyt R, Vega F Jr, Zurawski SM, Zurawski G, de Vries JE. An interleukin 4 (IL-4) mutant protein inhibits both IL-4 or IL-13-induced human immunoglobulin G4 (IgG4) and IgE synthesis and B cell proliferation: support for a common component shared by IL-4 and IL-13 receptors. J Exp Med. 1993;178(6):2213-8. https://doi.org/10.1084/jem.178.6.2213
  42. Lee IS, Dat NT, Cai XF, Shen G, Kim YH. Inhibitory effects of natural products against NFAT (nuclear factor of activated T cells) transcription factor. Korean J Pharmacogn. 2003;34(2):150-5.
  43. Liebmann C. Regulation of MAP kinase activity by peptide receptor signalling pathway: paradigms of multiplicity. Cell Signal. 2001;13(11):777-85. https://doi.org/10.1016/S0898-6568(01)00192-9
  44. Cheung PF, Wong CK, Ho AW, Hu S, Chen DP, Lam CW. Activation of human eosinophils and epidermal keratinocytes by Th2 cytokine IL-31: implication for the immunopathogenesis of atopic dermatitis. Int Immunol. 2010;22(6):453-67. https://doi.org/10.1093/intimm/dxq027
  45. Park GY, Lee SH, Hwangbo B, Yim JJ, Lee CT, Kim YW, Han SK, Shim YS, Yoo CG. Pro-inflammatory cytokine expression through NF-${\kappa}B$/$I{\kappa}B$ pathway in lung epithelial cells. Tuberc Respir Dis. 2000;49(3):332-42. https://doi.org/10.4046/trd.2000.49.3.332
  46. Pahl HL. Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene. 1999;18(49):6853-66. https://doi.org/10.1038/sj/onc/1203239
  47. Abul K. Abbas, Andrew H. Lichtman. Basic Immunology, 3th ed. Seoul: Seoul: Epublic Publishing Co. 2009:104-11,205-12.