The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology (한방안이비인후피부과학회지)

The Society of Korean Medicine Ophthalmology, Otolaryngology & Dermatology (대한한방안이비인후피부과학회)
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The Effect of Moutan Cortex on Pro-inflammatory Cytokines through NF-κB & MAPKs pathway in HMC-l

목단피의 NF-κB와 MAPKs 억제를 통한 Pro-inflammatory Cytokines 활성에 미치는 효과

  • 박주호 (경원대학교 한의과대학 안이비인후피부과학교실) ;
  • 김경준 (경원대학교 한의과대학 안이비인후피부과학교실)
  • Published : 2009.08.25
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Abstract

Objective : Moutan Cortex (the root bark of Paeonia suffruticosa Andr.) is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how MC(Moutan Cortex) affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted MC on the mast cell-mediated inflammatory responses. Method : We observed the effect of MC on compound 48/80-induced histamine release of rat peritoneal mast cells and the effect of administering MC on PCA in rat. We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of MC. The TNF-$\alpha$ protein levels were analysised by Western blot. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-$\kappa$B, phospho-I$\kappa$B and MAPKs were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by luciferase assay. Result : 1. Enzyme immunoassay indicated that MC suppressed histamine secretion of rat peritoneal mast cells. 2. In PCA dependent on IgE, MC had anti-allergic effect of the internal surface of rat skin. 3. Western blot indicated that MC decreased TNF-$\alpha$ protein levels. 4. ELISA indicated that MC decreased TNF-$\alpha$, IL-6 but MC had no significant effect on IL-8 in HMC-1 cells. 5. RT-PCR indicated that MC decreased TNF-$\alpha$, IL-8 but MC had no significant effect on IL-6 in HMC-l cells. 6. Western blot indicated that MC suppressed the induction of MAPKs, NF-$\kappa$B & phospho-I$\kappa$B activity in HMC-1 cells. 7. Luciferase assay indicated that MC suppressed the PMA plus A23187-induced NF-$\kappa$B promoting activityin HMC-1 cells. Conclusion : In this study, we have found that MC is an inhibitor of NF-$\kappa$B, MAPKs & cytokines on the mast cell-mediated inflammatory responses.

Keywords

References

  1. 전국 한의과대학 피부외과학 교재편찬위원회. 韓醫皮膚外科學. 부산:선우. 2007:79-81.
  2. Peter P. 면역학(The Immune System 2nd edition). 서울:라이프사이언스. 2006:7, 236-7, 307-37.
  3. 西田榮介, 大野茂男. 정보전달. 서울:월드사이언스. 2004;93-117, 126.
  4. Beg AA, Baldwin AS Jr. The I kappa B protein: multifunctional regulators of Rel/NF-kappa B transcription factor. Genes Dev. 1993; 7(11):2064-70. https://doi.org/10.1101/gad.7.11.2064
  5. Palombella VJ, Rando OJ, Goldberg AL, and Maniatis T. The ubiquitinproteasome pathway is required for processing the NF-$\kappa$B1 precursor protein and the activation of NF-$\kappa$B. Cell. 1996;78(5):773-85.
  6. Kuprsh DV, Udalova IA, Turetskaya RL, Rice NR, Nedospasov SAC. Convserved kappa B element located downstream of the tumor necrosis factor alpha gene: distinct NF-kappa B binding pattern and enhancer activity in LPS activated murine macrophages. Oncogene. 1995;11:97-106.
  7. Galien R, Evans HF, Garcia T. Involvement of CCAAT/ enhancerbinding protein and nuclear factor-kappa B binding site in interleukin-6 promoter inhibition by estrogens. Mol Endocrinol. 1996;10:713-22. https://doi.org/10.1210/me.10.6.713
  8. 全國韓醫科大學 本草學敎授 共編著. 本草學. 서울:永林社. 1992:193-95.
  9. 식품의약품안전청. 대한약전외한약(생약)규격집. 한국의약품수출입협회. 2000:155-6.
  10. 안덕균 저. 한국 본초도감. 서울:교학사. 1998: 366.
  11. 李時珍. 本草綱目. 北京:人民衛生出版社. 1982:852-4.
  12. 國家中醫藥管理局 <中華本草> 編委會. 中華本草. 上海:上海科學技術出版社. 1999;3:528-35.
  13. Hachisuka H, Kusuhara M, Higuchi M, Okubo K, Sasai Y. Purification of rat cutaneous mast cells with Percoll density centrifugation. Arch Dermatol Res. 1988;280:358-62. https://doi.org/10.1007/BF00426614
  14. 대한 소아알레르기 및 호흡기학회편. 소아 알레르기 호흡기학. 서울:군자출판사. 2005;11-46.
  15. Ishizaka T, Soto CS, Ishizaka K. Mechanical of passive sensitization-III. Number of IgE molecules and their recepter sites on human basophil granulocytes. J Immunol. 1973;111:500-11.
  16. Petersen LJ, Mosbech H, Skov P. Allergen induced histamine release in intact human skin in vivo assessed by skin microdialysis technique. Characterization of factors in fulencing histamine releasability. J Allergy Clin Immunol. 1996;97:672. https://doi.org/10.1016/S0091-6749(96)70313-5
  17. Janeway CA, Travers P, Walport M, Capera JD. in Immunobiology: The Immune System in Health and Disease (Current Biology Publication, London). 1999;4th Ed.:461-88.
  18. Arend WP, Dayer JM. Inhibition of the production and effects of interleukin-1 and tumor necrosis factor alpha in rheumatoid arthritis. Arthritis Rheum. 1995;38(2):151-60. https://doi.org/10.1002/art.1780380202
  19. Erchler WB, Keller ET. Age-associated increased interreukin-6 gene expression, late-life diseases, and frailty. Annu Rev Med. 2000;51:245-70. https://doi.org/10.1146/annurev.med.51.1.245
  20. Janeway C.A. Jr, Travers P, Walport M, Shlomchik M. 면역생물학(Immunobiology 6th). 서울:라이프사이언스. 2005;801.
  21. Harvey L, Arnold B, Paul M, Chris AK, Monty K, Matthew PS, Lawrence Z, James D. 분자세포생물학(Molecular Cell Biology 5th edition). 서울:월드사이언스. 2006:601-2.
  22. Barnes PJ, Karin M. Nuclear factor-$\kappa$B: a pivotal transcription factor in chronic inflammatory disease. N Engl J Med. 1997;336:1066-71. https://doi.org/10.1056/NEJM199704103361506
  23. Geoffrey M.Cooper. 분자세포생물학(The Cell). 서울:한우리. 2002;491.
  24. Parra V, Gadina M, Kovarova M, et al. Fyn kinase initiates complementry signals required for IgE-dependent mast cell degranulation. Nat. Immunol. 2002;3:741-48. https://doi.org/10.1038/ni817
  25. Cohen MP, Shea E, Chen S, Shearman CW. Glycated albumi increase oxidative stress, activates NF-kappa B and extracellular signal-rerulated kinase(ERK), and stimulates ERK-dependent transforming growth factor-beta 1 production in macrophage RAW cells. J Lab Clin Med. 2003;141(4):242-9. https://doi.org/10.1067/mlc.2003.27
  26. Dean JLE, Brook M, Clark AR, Saklatavala J. p38 mitogen activated protein kinase regulates cycloxygenase-2 mRNA stability and transcription in lipopolysaccaridetreated human monocytes. J Biol chem. 1999;274:264-9. https://doi.org/10.1074/jbc.274.1.264
  27. Lin A, Minden A, Martinetto H, Claret FX., Lango-Carter C, Mercurio F, Johnson GI, Karin M. Identification of dual specificity kinase that activates the jun kinases and p38-mpk2. Science 1995;268:286-90. https://doi.org/10.1126/science.7716521
  28. Wang C, Deng L, Hong M, Akkaraju GR, Inoue J, Chen ZJ. TAK1 is ubiquitindependent kinase of MKK and IKK. Nature 2001;412:346-51. https://doi.org/10.1038/35085597
  29. Song HY, Regnier CH., Kirschning CJ, Goeddel DV, Rothe M. Tumor necrosis factor(TNF)-mediated kinases cascades: Bifurcation of Nuclear Factor-$\kappa$B and c-Jun N terminal kinases(JNK/SARK) pathways at TNF receptor-assoiated factor 2. Proc Natl Acad Sci USA 1997;94:9792-96. https://doi.org/10.1073/pnas.94.18.9792
  30. Baud V, Liu ZG, Bennette B, Suzuki N, Xia Y, Karin M. Signaling by proinflammatory cytokines: oligomerization of TRAF2 and TRAF6 is sufficient for JNK and IKK activation and target gene activation via an amino-terminal effector domain Genes derv 1999;13:1297-308. https://doi.org/10.1101/gad.13.10.1297
  31. Ono K, Han J. The p38 signal transduction pathways : activation and function. Cell signal. 2000;12:1-13. https://doi.org/10.1016/S0898-6568(99)00071-6
  32. 송계용, 지제근, 함의근. 핵심병리학. 서울:고려의학. 1998:193-202.
  33. Rao A. NF-ATp: a transcription factor required for the co-ordinate induction of several cytokine genes. Immunol Today. 1994;15:274-81. https://doi.org/10.1016/0167-5699(94)90007-8
  34. Firrini E, Marchisio PC, Scupoli MT, Poffe O, Tagliabue E, Brentegani M, Colombatti M, Santini F, Tridente G, Ramarli D. Adhesion of immature and mature T cells induces in human thymic epithelial cells(TEC) activation of IL-6 gene transcription factors (NF-kappaB and NF-IL6) and IL-6 gene expression: role of alpha3beta1 and alpha6beta4 integrins. Dev Immunol. 2000;7:195-208. https://doi.org/10.1155/2000/48239
  35. 노영득. 牧丹皮의 항염효과에 관한 연구. 익산. 원광대학교 대학원: 한의학과. 2004.
  36. 오중환. 목단피와 Paeonol의 항염증작용 및 항염증 작용 및 TNF-$\alpha$,iNOS에 생성억제효과. 경원대학교 대학원: 한의학과. 2005.