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The Effects of Forsythiae Fructus n-BuOH Fraction on Atopic Dermatitis

연교(連翹) n-BuOH 분획물의 아토피 피부염 억제 효과

  • Lee, Jin Hwa (Department of Pediatrics, College of Korean Medicine, Daejeon University) ;
  • Han, Jae Kyung (Department of Pediatrics, College of Korean Medicine, Daejeon University) ;
  • Kim, Yun Hee (Department of Pediatrics, College of Korean Medicine, Daejeon University)
  • 이진화 (대전대학교 한의과대학 소아과학교실) ;
  • 한재경 (대전대학교 한의과대학 소아과학교실) ;
  • 김윤희 (대전대학교 한의과대학 소아과학교실)
  • Received : 2016.07.22
  • Accepted : 2016.08.16
  • Published : 2016.08.31

Abstract

Objectives Previous studies have found out that Forsythiae Fructus (FF) extracts have anti-atopic activities by in vitro experiment. In order to understand more about FF extracts' benefit, we subdivided FF extracts depending on systematic fractionation method by using Methylene chloride (MC), Ethyl acetate (EtOAc), n-BuOH and n-hexane (n-Hx). This study is designed to examine the effect of FF fractions on the PMA- ionomycin-induced activation of RBL-2H3 mast cell lines in vitro and on the DNCB-induced activation of NC/Nga mice in vivo. Methods For this study, we examined IL-4, IL-13 production by ELISA analysis, IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$ mRNA expression by real-time PCR and manifestations of AP-1 and MAPKs transcription factors by western blotting in vitro. Through in vitro experiment, we selected FF n-BuOH fraction that seems the best effective in atopic dermatitis then induced it on NC/Nga mice by DNCB. We measured mice's WBC, eosinophil and neutrophil in heart blood, IL-4, IL-5, IFN-${\gamma}$ in the spleenocyte culture supernatant, the absolute cell numbers of CD4+, CD8+, B220+CD23+, CD3+CD69+ and Gr-1+CD11b+ in the PBMCs, ALN and dorsal skin, IL-5, IL-13, IL-31, IL-31RA in the dorsal skin by real-time PCR and the distribution of immune cells by H&E on dorsal skin and ANL and toluidine blue staining on dorsal skin. Results FF n-BuOH fraction suppressed IL-4, IL-13 production and mRNA expression of IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$. Results from the western blot analysis showed that FF n-BuOH fraction reduced the activation of the mast cell specific transduction factors involved in AP-1 by suppressing JNK and ERK phosphorylation. In the gross, atopic dermatitis induced by DNCB in NC/Nga mice were improved by oral administration of FF n-BuOH fraction. Oral FF n-BuOH fraction also decreased the level of IgE in mice's serum and the level of IL-4 and IL-5 in the spleenocyte culture supernatant, cell numbers of CD8+, B220+CD23+ in the PBMCs, CD4+ in the ALN and CD4+, Gr-1+CD11b+ in the dorsal skin and suppressed mRNA expression of IL-5, IL-13, IL-31, IL-31RA in the dorsal skin. Histological examination showed that infiltration levels of immune cells in atopic dermatitis induced NC/Nga mice were improved by FF n-BuOH fraction. Conclusions FF n-BuOH fraction can reduce pruritus by suppressing IL-31, IL-31RA secretion and modulate molecular mediators and immune cells associated with atopic dermatitis induced in NC/Nga mice which may have played a significant role in recovering atopic dermatitis symptoms.

Keywords

References

  1. Ahn HS. Hongchangui pediatrics, 9th edition, Seoul: Miraen. 2010:1142.
  2. Park YM. Advances in the pathophysiology of atopic dermatitis. Pediatr Allergy Respir Dis. 2006;16(3):189-96.
  3. Mudde GC, Reijsen FC, Boland GJ, Bruijnzeel PL, Bruijnzeel-K. Allergen presentation by epidermal Langerhans' cells from patients with atopic dermatitis is mediated by IgE. Immunol. 1990;69(3):335-41.
  4. Markus G, Carla A, Bruijnzeel-K, Erwin S, Theo T, Alice G. Langeveld-W, Thomas R, Jean K. A role for Th1 and Th2 cells in the immunopathogenesis of atopic dermatitis. Immunol. 1998;19(8):359-61.
  5. Reijsena FC, Bruijnzeel-K, Kalthoff F,. Maggi E, Romagnani S, Westlanda J, Mudde GC. Skin-derived aeroallergen-specific T-cell clones of Th2 phenotype in patients with atopic dermatitis. J Allergy Clin Immunol. 1992;90(2):184-93. https://doi.org/10.1016/0091-6749(92)90070-I
  6. Foster PS, Martinez-Moczygemba M, Huston DP, Corry DB. Interleukins -4, -5, and -13; emerging therapeutic targets in allergic disease. Pharmacol Ther. 2002;94(3): 253-624. https://doi.org/10.1016/S0163-7258(02)00220-6
  7. Matsubara M, Masaki S, Ohmori K, Karasawa A, Hasegawa K. Differential regulation of IL-4 expression and degranulation by anti-allergic olopatadine in rat basophilic leukemia (RBL-2H3) cells. Biochem Pharmacol. 2004;67(7):1315-26. https://doi.org/10.1016/j.bcp.2003.12.008
  8. Hanifin JM, Rajka G. Diagnostic features of atopic dermatitis. Acta Derm Venereol. 1980;92:44-7.
  9. Yoon JH, Bae JH, Difficulties and needs of parents raising young children with atopic dermatitis. Eco-Early Child Edu. 2013;12(1):113-37.
  10. Harri A, Marie-C, Stephan M, Juliane R, Martin S, Thomas K, Hoffmann, Thomas R, Albert Z, Bernhard H. IL-31: A new link between T cells and pruritus in atopic skin inflammation. J Allergy Clin Immunol. 2006;177(2):411-7.
  11. Kim S, Kim HJ, Yang HS, Kim EG, Huh IS, Yang JM. IL-31 serum protein and tissue mRNA levels in patients with atopic dermatitis. Ann Dermatol. 2011; 23(4):468-73. https://doi.org/10.5021/ad.2011.23.4.468
  12. Ulrike R, Katja W, Manuela B, Sonja S, Bettina W, Alexander K, Thomas W. Correlation of IL-31 serum levels with severity of atopic dermatitis. J Allergy Clin Immunol. 2006;122(2):421-3. https://doi.org/10.1016/j.jaci.2008.05.047
  13. Grimstad Q, Sawanobori Y, Vestergaard C, Bilsborough J, Olsen UB, Larsen C, Matsushima K. Anti interleukin 31 antibodies ameliorate scratching behaviour in NC/Nga mice; a model of atopic dermatitis. Experiment Dermatol. 2009;18(1):35-43. https://doi.org/10.1111/j.1600-0625.2008.00766.x
  14. Ju YS. Ungok herbology. Jeonju: Woosuk Press. 2013:494-7.
  15. Herbology Association of Korea's Colleges of Oriental Medicine. Textbook of herbology, 1st ed. Seoul: Younglimsa. 1991:199.
  16. Lee JH, Han JK, Kim YH. The suppressive effect of Th2 cytokines expression and the signal transduction mechanism in MC/9 mast cells by Forsythiae Fructus extracts. J Pediatr Korean Med. 2014;28(3):31-46. https://doi.org/10.7778/jpkm.2014.28.3.031
  17. Nam JB, Lee MH, Choi HY, Sohn NW, Kang H. Effect of Forsythiae Fructus ethanol extract on inflammatory cytokine production and cellular signaling pathways in mouse macrophages. Korean J Herbol. 2012;27(1):59-64.
  18. Kim SY, Park YK. Ethanol extract of Forsythiae Fructus inhibits the production of inflammatory mediators in LPS-stimulated BV-2 microglial cells. Korean J Herbol. 2008;23(3):93-102.
  19. Lee JM, Choi SW, Cho SH, Phee SJ. Effect of Forsythia Viridissima extracts on antiosidative system and lipid peroxidation of liver in rats fed high-cholesterol die. Koran J Nutr Health. 2003;36(10):990-6.
  20. Jung YJ, Jeon YS, Kim HJ, Kang SK, Kim YK, Kim NS. Anti-allergic and anti-inflammatory effect of Leonurus Sibiricus seed ethyl acetate fractions. Yakhak Hoeji. 2014;58(5):294-9.
  21. Yang CH, Lee YM, Cho KY, Bae KH, Chung CP. Effect of Zizyphi Fructus extract on the biological activity of Gingival Fibroblast. J Korean Periodontol. 1994; 24(1):144-54.
  22. Lee EJ, Yang SA, Choi HD, Im HG, Whag K, Lee IS. Comparison of Gingerols in various fractions and the antioxidant effects of supercritical fluid extracts from Ginger. Korean J Food Sci Technol. 2011;43(4):469-74. https://doi.org/10.9721/KJFST.2011.43.4.469
  23. Jung BM, Park JA, Bae SJ. Growth inhibitory and quinone reductase induction activities of Salicornia herbacea L. fractions on human cancer cell lines in vitro. J Korean Soc Food Sci Nutr. 2008;37(2):148-53. https://doi.org/10.3746/jkfn.2008.37.2.148
  24. Kim JH. Recent advances in atopic dermatitis. Allergy Asthma Respir Dis. 2004;14(1):12-23.
  25. Kim CH, Lee SI, Lee SY, Choi SY, Lee JS, Kim KE. Nationwide study for epidemiological change of atopic dermatitis in school aged children between 1995 and 2000 and kindergarten aged children in 2003 in Korea. Allergy Asthma Respir Dis. 2003;13(4):227-37.
  26. Greer FR, Sicherer SH, Burks AW. Effect of early nutritional interventions on the development of atopic disease in infants and children; the role of maternal dietary restriction, breastfeeding, timing of introduction of complementary foods, and hydrolyzed formulas. Pediatr. 2008;111:608-16.
  27. Vander H, Klip H, Brand PI. Risk of developing asthma in young children with atopic eczema; a systemic review. J Allegy Clin Immunol. 2007;120:565-9. https://doi.org/10.1016/j.jaci.2007.05.042
  28. Lee JH, Kim KH, Kim MN, Kim JW, No YS, Park YL, Park CW, Lee KH, Lee AY, Jo SH, Choi JH. The treatment guideline of Korean atopic dermatitis. Korean J Dermatol. 2006;44(8):907-13.
  29. Katayama I, Kohno Y, Akiyama K, Ikezawa Z, Kondo N, Tamaki K, Kouro O. Japanese guideline for atopic dermatitis. Allergol Int. 2011;60:205-20. https://doi.org/10.2332/allergolint.11-RAI-0333
  30. Schacke H, Docke WD, Asadullah K. Mechanisms involved in the side effects of glucocorticoids. Pharmacol Ther. 2002;96:23-43. https://doi.org/10.1016/S0163-7258(02)00297-8
  31. Dowell SF, Bresee JS. Severe varicella associated with steroid use. Pediatr. 1993;92:223-8.
  32. Kim SC, Ahn KJ, Hann SK, Kim JW, Sung KJ, Kye YC, Kim NI, Cho KH, Kim KJ, Youn JI. Clinico-epidemiologic study on the abuse, misuse, and adverse effects of topical dermatologic drugs. Korean J Dermatol. 2003;41(9):1129-35.
  33. Jin HW, Jang HS, Jang BS, Jo JH, Kim MB, Oh CG, Gwon KS, Gwon YY. The use of alternative medicine and management in children with atopic dermatitis. Korean J Dermatol. 2005;43(7):903-11.
  34. Kim SH. The use of oriental and complementary alternative medicine in children with atopic dermatitis. University Master of Cha Medical University. 2007.
  35. 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.
  36. Schulz F, Marenholz I, Foster-Holst R, Chen C, Sternjak A, Baumgrass R. A common haplotype of the IL-31 gene influencing gene expression is associated with nonatopic eczema. J Allergy Clin Immunol. 2007;120: 1097-102. https://doi.org/10.1016/j.jaci.2007.07.065
  37. Zhang Q, Prabhakar P, Zhou Q, Liu QS, Gao WD. Structures and biological functions of IL-31 and IL-31 receptors. Cytokine Growth Factor Rev. 2008; 19(2008):347-56. https://doi.org/10.1016/j.cytogfr.2008.08.003
  38. Cornelissen C, Luscher-Firzlaff J, Baron JM, Luschera B. Signaling by IL-31 and functional consequences. Eur J Cell Biol. 2011;91(2012):552-66.
  39. Ha NL, Lee JC, Kim KB. Recent research on treatments of atopic dermatitis in Japan. Diet Kampo Med. 2012;26(1):16-35.
  40. Min DL, Park EJ, Kang KH. Review of clinical and experimental studies on external application treatment for atopic dermatitis in the Korean literature. J Pediatr Korean Med. 2013;27(1):36-49. https://doi.org/10.7778/jpkm.2013.27.1.036
  41. Yoon JY, Han JK, Kim YH. Effect of Kami-chungsimyeunjatang on atopic dermatitis-like skin lesions induced in NC/Nga mice by mite antigen stimulation. J Korean Orient Pediatr. 2007;21(1):87-116.
  42. Ko MJ, Baek JH. A clinical study on the effect of Hwangryunhaedock-tang on atopic dermatitis. J Korean Orient Pediatr. 2012;26(4):51-60. https://doi.org/10.7778/jpkm.2012.26.4.051
  43. Park JM, Chae JW. Effects of Aurantii Immaturus Fructus (AI) on atopic dermatitis (AD) induced by DNCB in mice. J Pediatr Korean Med. 2015;29(1):27-43. https://doi.org/10.7778/jpkm.2015.29.1.027
  44. Yeo EJ, Han JK, Kim YH. Inhibitory studies of Scrophulariae Radix on development of dermatitis in NC/Nga mice. J Korean Orient Pediatr. 2007;21(2): 13-34.
  45. Liebmann C. Regulation of MAP kinase activity by peptide receptor signalling pathway; paradigms of multiplicity. Cell Signal. 2001;13:777-85. https://doi.org/10.1016/S0898-6568(01)00192-9
  46. Whitmarsh AJ, Davis RJ. Transcription factor AP-1 regulation by mitogen-activated protein kinase signal transduction pathways. J Mol Med. 1996;74:589-607. https://doi.org/10.1007/s001090050063
  47. Lewin I, Nechushtan H, Ke Q, Razin E. Regulation of AP-1 expression and activity in antigen-stimulated mast cells; the role played by protein kinase C and the possible involvement of Fos interacting protein. Blood. 1993;82:3745-51.
  48. Yun YG, Hwang JM, Kim HR, Jang SU. Mouse models of atopic dermatitis for drug discovery from medicinal plants. Korean J Orient Med Prescription. 2007;150: 145-61.
  49. Venkayya R, Lam M, Willkom M, Grunig G, Corry DB, Erle DJ. The Th2 lymphocyte products IL-4 and IL-13 rapidly induce airway hyperresponsiveness through direct effects on resident airway cells. Am J Respir Cell Mol Biol. 2002;26:202-8. https://doi.org/10.1165/ajrcmb.26.2.4600
  50. Kim HM, Eom JY, Jeong HJ, Hong SH. Immune and allergic. Seoul: Shinil Books. 2008:168, 171, 222.
  51. Humbert M, Grant JA, Taborda-Barata L, Durham SR, Pfister R, Menz G. High-affinity IgE receptor (Fcepsiolon RI)-bearing cells in bronchial biopsies from atopic and nonatopic asthma. Am J Respir Crit Care Med. 1996;153:1931-7. https://doi.org/10.1164/ajrccm.153.6.8665058
  52. Toru H, Pawankar R, Ra C, Yata J, Nakahata T. Human mast cells produce IL-13 by high-affinity IgE receptor cross-linking; enhanced IL-13 production by IL-4-primed human mast cells. J Allergy Clin Immunol. 1998;102(3): 491-502. https://doi.org/10.1016/S0091-6749(98)70140-X
  53. Kim JW. Atopic dermatitis at allergic and immunological aspects. J Korean Dermatol Assoc. 2003;41(6): 687-9.
  54. Abbas Abul K, Lichtman Andrew H, Pillai Shiv. Cellular and Molecular Immunology 6th ed. Philadelphia: Saunders. 2007:304-15.
  55. Mehrotra PT, Wu D, Crim JA, Mostowski HS, Siegel JP. Effects of IL-12 on the generation of cytotoxic activity in human CD8+ T lymphocytes. J Immunol. 1993; 151(5):2444-52.
  56. Zdolsek HA, Ernerudh J, Holt PG, Nilsson J, Bjorksten B. Expression of the T-cell markers CD3, CD4 and CD8 in healthy and atopic children during the first 18 months of life. Arch Allergy Immunol. 1999; 119:6-12. https://doi.org/10.1159/000024169
  57. Kim SJ. Immunology 1st ed. Seoul: Koryo medicine. 1994:150-3, 206.
  58. Rich Robert R, Fleisher Thomas T, Shearer William T, Schroeder H. Clinical immunology. New York: Mosby. 2001;chap4:13, chap6:18.
  59. Dorshkind K, Narayanan R, Landreth KS. Regulatory cells and cytokines involved in primary B lymphocyte production. Adv Exp Med Biol. 1992;323:119-23.
  60. Askenasea Philip W, Kawikovaa I, Paliwala V, Akahira-Azumaa M, Gerardc C, Huglib T, Tsujid. A new paradigm of T cell allergy; requirement for the B-1 cell subset. Int Arch Allergy Immunol. 1999;118: 145-9. https://doi.org/10.1159/000024052
  61. Bronte V, Wang M, Overwijk WW, Surman DR, Pericle F, Rosenberg SA, Restifo NP. Apoptotic death of CD8+ T lymphocytes after immunization: induction of a suppressive population of Mac-1+/Gr-1+cells. J Immunol. 1998;161(10):5313-20.
  62. Angulo I, Heras FG, Garcia-Bustos JF, Gargallo D, Munoz-Fernandez A, Fresno M. Nitric oxide-producing CD11b (+)Ly- 6G (Gr-1) (+)CD31 (ER-MP12) (+) cells in the spleen of cyclophosphamide- treated mice: implications for T-cell responses in immuno-suppressed mice. Blood. 2000;95(1):212-20.
  63. Park EH, Yoo JH, Gim SB, Lee YK, Kim DH. Effect of Hyunggaeyunkyotangbalhyobang (HYBH) on atopic dermatitis in NC/Nga mice model. J Orient Med Daejeon University. 2011;19(2):65-83.
  64. Han DS, Han JK, Kim YH, Effects of SPDJTK (SoPungDoJeokTangKami) and concurrent administration of AJ (Atopy cream, Jawoongo) plus SPDJTK on atopic dermatitis-like skin lesions in NC/Nga mouse induced by BMAC. J Korean Orient Pediatr. 2010;24(1): 9-35.
  65. Lee YS, Han JK, Kim YH, The effects of Dictamni Radicis cortex extracts on dncb induced atopic dermatitis-like skin lesions of NC/Nga mouse. J Pediatr Korean Med. 2014;28(4):1-29. https://doi.org/10.7778/JPKM.2014.28.4.001

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