The Journal of Pediatrics of Korean Medicine (대한한방소아과학회지)

The Association of Korean Oriental Pediatrics (대한한방소아과학회)
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The Suppressive Effect on Th2 Cytokines Expression and the Signal Transduction Mechanism in MC/9 Mast Cells by PRAL

MC/9 비만세포에서 행인(杏仁) 추출물의 Th2 cytokine 발현 억제 효과 및 신호전달 기전 연구

  • Kang, Ki Yeon (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)
  • 강기연 (대전대학교 한의과대학 소아과학교실) ;
  • 한재경 (대전대학교 한의과대학 소아과학교실) ;
  • 김윤희 (대전대학교 한의과대학 소아과학교실)
  • Received : 2014.04.25
  • Accepted : 2014.05.20
  • Published : 2014.05.31
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Abstract

Objectives PRAL (Prunus armniaca Linne Var) is a herbal formula in Oriental Medicine, known for its anti-inflammatory and anti-allergenic properties. However, its mechanism of action and the cellular targets have not yet been found enough. The purpose of this study is to investigate the effects of PRAL on Th2 cytokines expression in MC/9 mast cells. Methods The effect of PRAL was analyzed by ELISA, Real-time PCR, Western blot in MC/9 mast cells. mRNA levels of GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ were analyzed with Real-time PCR. Levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results PRAL inhibited GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ mRNA expression in a dose dependent manner. GM-CSF, IL-4, IL-5 mRNA expression were inhibited significantly in comparison to DNP-IgE control group at concentration of 100 ${\mu}g/ml$ and IL-6, IL-13, TNF-${\alpha}$ mRNA expression were inhibited at concentration of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$. PRAL also inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group in a dose dependent manner. IL-13 production was inhibited at a concentration of 200 ${\mu}g/ml$, 400 ${\mu}g/ml$ and MIP-$1{\alpha}$ was inhibited at a concentration of 100 ${\mu}g/ml$, 200 ${\mu}g/ml$, 400 ${\mu}g/ml$. Western blot analysis of transcription factors involving Th2 cytokines expression revealed prominent decrease of the mast cell specific transcription factors including NFAT-1, c-Jun as well as NF-${\kappa}B$ p65 but not NFAT-2 and c-Fos. Conclusion These results indicate that PRAL has the effect of suppressing Th2 cytokines production in the MC/9 mast cells. These data represent that PRAL potentiates therapeutic activities to the allergic disease by regulating Th2 cytokines in the MC/9 mast cells.

Keywords

References

  1. Abbas Ak, Lichtman AH, Pillai S. Cellular & Molecular Immunology. 6th ed. Seoul: Epublic Publishing Co. 2008:451-71.
  2. An HS. Textbook of pediatrics. 9th ed. Seoul: Korea Textbook Publishing Co. 2010:1104-21.
  3. Seo SH. Prevalence of allergic diseases in Korean children, 2010. Public health weekly report, KCDC. 2011;4(24): 425-31.
  4. Ju CY, Heo TS, Han DH, Jung HJ, Jung SK, Rhee HK. A questionnaire analysis about the understanding allergic disease and the attitude toward oriental medical treatment. Korean J Orient Int Med. 2000;21(4):565-73.
  5. Hanuigwadaehak Bonchohak Pyeonchanwiwonhoe. Bonchohak. Seoul: Yeongrimsa Publishing Co. 2005: 515-6.
  6. Jeon YK. Kook YB, Kim YK, Yun YG. A study on application of semen armeniacae amarum main blended prescription from Donguibogam. Korean J Orient Med Prescription. 2004;12(2):31-46.
  7. Hwang DR, Kang YS, Kim SS, Kim DH, Shin MK, Son HJ. Studies on the allergic asthma effect of semen armeniaceae amarum. Korean J Herbology. 2003;18(2) 201-8.
  8. Ju CY, Jung HJ, Jung SK, Rhee HK. Armeniacae amarum semen contributes to the chemotaxis of eosinophils and secretion of chemokines in A549 human epithelial cells. Korean J Orient Int Med. 2006;27(1):208-20.
  9. Chung W, Jung HJ, Jung SK, Rhee HK. The effects of armeniacae arnarum semen and platycodi radix on IL-4, IL-5, IL-6 in asthma model. Korean J Orient Int Med. 2000;21(1):31-8.
  10. Kim YS, Song CH. Study on the anti-allergic effect of armeniacae semen herbal acupuncture solution. Korean J Acupunct. 2007;24(2):151-62.
  11. Shin HW. The experimental study on the asthmasuppressive & immune response of armeniacae amarum semen herbal-acupuncture at ST36 in OVA-induced asthma mouse. Grad School Daejeon Univ. 2006.
  12. Munetaka I, Yoko M, Kazumi S, Yosuke O, Keiyu U. 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
  13. Han JH, Kim KY. Hanbangyakrihak. Seoul: Euisungdang Pub. 2004:405-6.
  14. An SK, Park BD, Won JH, Sung YO, Chun SH. Atopic dermatitis. Seoul: Pacific Pub. 2007:13-82.
  15. The Korean Academy of Asthma, Allergy and Clinical Immunology. Asthma and allergic disease. Seoul: Koonja Pub. 2002:13-116.
  16. Kim DM, Hong SP. Development of quantitative extraction method of amygdalin without enzymatic hydrolysis from tonin & kyonin by HPLC. Grad School Kyunghee Univ. 2003.
  17. Choi JY. The analgesic and anti-inflammatory effects of armenicae semen: Potential role of bradykinin and COX-II-mediated signaling. Grad School Kyunghee Univ. 2004.
  18. Kim JH, Kang TW, Park CB, Cha KJ, Ahn YK. Immunobiological studies on route of administration of amygdalin. Yakhak Hoeji. 1996;40(2):202-11.
  19. Park YM. Lecture of medical science : Advances in the pathophysiology of atopic dermatitis. Pediatr Allergy Respir Dis (Korea). 2006;16(3):189-96.
  20. Prieschl EE, Gouileux-Gruart V, Walker C, Hrrer 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.
  21. John A, Hamilton. GM-CSF in inflammation and autoimmunity. Trends Immunol. 2002;23(8):403-8. https://doi.org/10.1016/S1471-4906(02)02260-3
  22. Kim YK, Kim HJ, Kim WS, Park HJ, Moon G, Kim DW, Won JH. Inhibitory effect of Jaeumganghwa-tang on allergic inflammatory reaction. Korean J Orient Int Med. 2004;25(2):174-82.
  23. Koh DK, Yun JM, Lee TH. Effects of aconiti tuber on the change of interleukin-6 and TNF-$\alpha$ level induced by LPS I.C.V. injection in mice. Korean J Orient Med Prescription. 2004;12(1):195-208.
  24. Lee JE. Stimulation of osteoclastogenesis by enhanced levels of MIP-$1\alpha$ in BALB/c mice in vitro. Grad School Ulsan Univ. 2008.
  25. Lukacs NW. Role of chemokines in the pathogenesis of asthma. Nature Immunology Rev. 2001;1(2):108-16. https://doi.org/10.1038/35100503
  26. Donald NC. The role of MIP-1 alpha in inflammation and hematopoiesis. J Leukoc Biol. 1996;59(1):61-6. https://doi.org/10.1002/jlb.59.1.61
  27. Lee IS, Nguyen TD, Xing FC, Guanghai S, 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.
  28. Park JC. Role of casein kinase in the regulation of NFAT function. KSBMB News. 2000;20(2):1-7.
  29. Monticelli S, Solymar DC, Rao A. Role of NFAT proteins in IL-13 gene transcription in mast cells. J Biol Chem. 2004;279(35):36210-8. https://doi.org/10.1074/jbc.M406354200
  30. Zenz R, Eferl R, Scheinecker C, Redlich K, Smolen J, Schonthaler HB, Kenner L, Tschachler E, Wagner EF. Activator protein 1 functions in inflammatory bone and skin disease. Arthritis Res Ther. 2008;10(1):201.
  31. Lee KJ, Kim BK, Kil KJ. Suppressive effects of Morus alba Linne Root Bark(MRAL) on activation of MC/9 mast cells. Korean J Herbology. 2013;28(1):33-42. https://doi.org/10.6116/kjh.2013.28.1.33
  32. Rooney JW, Hoey T, Glimcher LH. Coordinate and cooperative roles for NFAT and AP-1 in the regulation of the murine IL-4 gene. Immunity. 1995;2(5):473-83. https://doi.org/10.1016/1074-7613(95)90028-4
  33. Schreck R, Albermann K, Baeurele PA. Nuclear factor kappa B: an oxidative stress-responsive transcription factor of eukaryootic cells. Free Radic Res Commun. 1992;17(4):221-37. https://doi.org/10.3109/10715769209079515
  34. Park GY, Lee SH, Hwangbo B, Im JJ, Lee CT, Kim YH, Han SG, Sim YS, Yu CG. Pro-inflammatory cytokine expression through NF-kB/IkB pathway in lung epithelial cells. Tuberc Respir Dis. 2000;49(3):332-42. https://doi.org/10.4046/trd.2000.49.3.332
  35. Sen R, Baltimore D. Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell. 1986;46(5):705-71. https://doi.org/10.1016/0092-8674(86)90346-6
  36. Pahl HL. Activators and target genes of Rel/NF-kappaB transcription factor. Oncogene. 1999;18(49):6853-66. https://doi.org/10.1038/sj.onc.1203239

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