Effects of KMS on the DNCB induced animal Model of Atopic Dermartitis

Article information

J Korean Med. 2019;40(1):63-77

doi : https://doi.org/10.13048/jkm.19006

Dea-Uk Song ¹, Ga-Yul Min ², SooYeon Hong ¹, Seo-Eun Lim ¹, Jung Huh ¹, Youngjoo Sohn ¹, Hyuk-Sang Jung ¹

¹Department of Anatomy, College of Korean Medicine, Kyung Hee University

²Department of pathology, college of oriental Medicine, Daejeon University

Correspondence to: 정혁상 (Hyuk-Sang Jung), 경희대학교 한의과대학 해부학교실, 서울특별시 동대문구 경희대로 26, 경희대학교 한의과대학 해부학교실, Tel: +82-2-961-9449, Fax: +82-2-961-9449, E-mail: jhs@khu.ac.kr

Received 2019 February 13; Revised 2019 March 12; Accepted 2019 March 12.

Abstract

Objectives

The purpose of this study is to examine the effect of water and fermentation extracts of KMS (Kami-Mihudeongsikjang-tang) on AD (atopic dermatitis). Additionally, by applying the fermentation extracts of KMS at the first sensitization and latency period, I investgated whether it could prevent AD.

Methods

In this study, to test the effect and preventive efficacy of KMSs on AD. DNCB-induced BALB/c mice of AD model was used. Through histological observation, epidermis and dermis thickness, the infiltration of eoshiphils and mast cells in epidermis and dermis were examined. We measured the serum level of IgE and IL-6, TNF-alpha, and the expressions of NF-κB and MAPK protein. In order to examine the effect of KMSs on keratinocyte, HaCaT cells were treated TNF-alpha and IFN-gamma to induce an inflammatory response. The KMSs were applied at various concentration in the experimental group. We investigated TARC expression.

Results

The treatment groups were reduced epidermis and dermis thickness, inhibited the infiltration of eosinophils and mast cells, reduced the serum level of IL-6. Moreover, sfKMS group reduced serum level of TNF-alpha, inhibited the protein expressions of NF-κB and the phosphoryllation of ERK, JNK and p38. Especially sfKMS-pre group were reduced the serum level of IgE, show significant inhibition on the protein expression of NF-κB and the phosphoryllation of ERK, JNK and p38. In the experiment on HaCaT cells, sfKMS group were reduced expression of TARC.

Conclusions

These result suggest that wKMS and sfKMS was effective in the treament on AD, and sfKMS would prevent AD.

Keywords: atopic dermatitis; prevention of atopic dermatitis; fermentation extract; Mihudeongsikjang-tang

Fig. 1

Experimental schedule for induction of atopic dermatitis. All topical applications were applied to the dorsal skin. The normal group was treated with a PBS.

Fig. 2

The effects of KMSs on epidermis and dermis thickness change in dorsal skin tissue of DNCB-induced BALB/c mice. (A) The thicknesses of epidermis and dermis were identified by H&E staining at magnification 100× (scale bar 200μm). (B) Measurement of epidermis thickness. (C) Data of dermis thickness. Each data represents the mean ± SEM. ##p < 0.01 compared with normal and *p < 0.05 compared with control. E: epidermis: D: dermis. KMS, kami-Mihudeongsikjang-tang. DNCB, 2,4-dinitrochlorobenzene.

Fig. 3

The effects of KMSs on mast cell infiltration in dorsal skin tissue of DNCB-induced BALB/c mice. (A) The infiltration of mast cell in epidermis and dermis lesions were examined by toluidine blue staining of skin section (magnification: 100′, scale bar: 200μm). (B) The number of mast cell. The infiltration of mast cell in three sites was counted. Each data represents the mean ± SEM. ##p < 0.01 compared with normal and **p<0.01 compared with control. KMS, kami-Mihudeong sikjang-tang. DNCB, 2,4-dinitrochlorobenzene.

Fig. 4

The effects of KMSs on eosinophil infiltration in dorsal skin tissue of DNCB-induced BALB/c mice. (A) The infiltration of eosinophil in epidermis and dermis lesions were examined by H&E staining of skin section (magnification: 100′ and 400′, scale bar: 200μm and 50μm). (B) The number of eosinophil. The infiltration of eosinophil in six sites was counted. Each data represents the mean ± SEM. ##p < 0.01 compared with normal and **p<0.01 compared with control. KMS, kami-Mihudeongsikjang-tang. DNCB, 2,4-dinitrochlorobenzene.

Fig. 5

The effects of KMS on NF-kB and MAPKs expressions was investigated by western blot. (A) The level of NF-kB (B) P-ERK, (C) P-JNK, (D) P-P38 expressions were examined by western blot. The expression level were measured by image J. Each data represents the mean SEM. ##p < 0.01 compared with normal and **p < 0.01, *p<0.05 compared with control. KMS, kami-Mihudeongsikjang-tang. NF-κB, nuclear factor kappa-light-chain-enhancer of activated B cells.. MAPK, Mitogen-activated protein kinase. ERK, extracellular signal-regulated kinase. T-ERK, total-ERK. P-ERK, phospho-ERK. T-JNK, total-JNK. P-JNK, phospho-JNK. T-p38, total-p38. P-p38, phospho-p38.

Fig. 6

The effects of KMSs on IgE and cytokines level in serum of DNCB-induced BALB/c mice. (A) The data of serum IgE (B) IL-6 (C) TNF-alpha. Each data represents the mean ± SEM. ##p< 0.01 compared with normal and **p < 0.01, *p <0.05 compared with control. KMS, kami-Mihudeongsikjang-tang. IgE, immunoglobulin E. IL-6, interleukin 6, TNF-alpha, tumor necrosis factor-alpha.

Fig. 7

The effects of KMS and sfKMS on TARC expressions. (A-B) ELISA, (C, D) RT-PCR. Each data represents the mean SEM. ##p < 0.01 compared with normal and *p < 0.05 compared with control. KMS, kami-Mihudeongsikjang -tang. TARC, thymus and activation-regulated chemokine. RT-PCR, Reverse transcription polymerase chain reaction.

Table 1

Composition of KMS.

References

1. Coca AF, Cooke RA. On the classification of the phenomena of hypersensitiveness. J Immunol 1923;8:163–182.

2. Nutten S. Atopic dermatitis: global epidemiology and risk factors. Ann Nutr Metab 2015;66(1):8–16.

3. Lee YU, Park SH, Nam EH, Chung TH, Hwang CY. Modulatory effects of PG102 (Actinidia arguta) on inflammatory cytokine mRNA expression in canine atopic dermatitis. Thai J Vet Med 2016;46(4):601–607.

4. Marco M, Heeney D, Binda S, Cifelli CJ, Cotter PD, Foligne B, et al. Health benefits of fermented foods: microbiota and beyond. Current Opinion in Biotechnology 2017;44:94–102.

5. Um YR, Lee JH, Ma JY. Quantitative analysis of marker substances in Solid fermented Angelicae Gigantis Radis by HPLC. Korean Journal of Oriental Medicine 2010;16(1):173–178.

6. Wickman M, Asarnoj A, Tillander H, Andersson N, Bergstrom A, Kull I, et al. Childhood-to -adolescence evolution of IgE antibodies to pollens and plant foods in the BAMSE cohort. J Allergy Clin Immunol 2014;133(2):580–582.

7. Westman M, Asarnoj A, Hamsten C, Wickman M, van Hage M. Windows of opportunity for tolerance induction for allergy by studying the evolution of allergic sensitization in birth cohorts. Seminars in Immunology 2017;30:61–66.

8. Peng W, Novak N. Pathogenesis of atopic dermatitis. Clinical & Experimental Allergy 2015;45:566–574.

9. Bieber T. Atopic Dermatitis. Ann Dermatol 2010;22:125–137.

10. Kasutani K, Fujii E, Ohyama S, Adachi G, Hasegawa M, Kitamura H, Yamashita N. Anti-IL-31 receptor antibody is shown to be a potential therapeutic option for treating itch and dermatitis in mice. Br J Pharmacol 2014;171:5049–5058.

11. Mu Z, Zhao Y, Liu X, Chang C, Zhang J. Molecular biology of atopic dermatitis. Clin Rev Allergy Immunol 2014;47:193–218.

12. Johansson SG, Bieber T, Dahl R, Friedmann PS, Lanier BQ, et al. Revised nomenclature for allergy for global use: Report of the Nomenclature Review Committee of the World Allergy Organization. J Allergy Clin Immunol 2004;113:832–836.

13. Ong PY, Leung DY. Immune dysregulation in atopic dermatitis. Curr Allergy Asthma Rep 2006;6:384–389.

14. Leung DY, Boguniewicz M, Howell M, Nomura I, Hamid QA. New insights into atopic dermatitis. J Clin Invest 2004;113:651–657.

15. Mosmann TR, Cherwinski H, Bond MW, Giedlin MA, Coffman RL. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol 1986;136(7):2348–2357.

16. Maggi E, Prete GD, Macchia D, Parronchi P, Tiri A, Chrétien I, et al. Profiles of lymphokine activities and helper function for IgE in human T cell clones. Eur J Immunol 1988;18:1045–1052.

17. Robinson DS, Hamid Q, Ying S, Tsicopoulos A, Barkans J, Bentley AM, et al. Predominant TH2-like bronchoalveolar Tlymphocyte population in atopic asthma. N Engl J Med 1992;326:298–304.

18. Chan LS, Robinson N, Xu L. Expression of interleukin-4 in the epidermis of transgenic mice results in a pruritic inflammatory skin disease: an experimental animal model to study atopic dermatitis. J Invest Dermatol 2001;117:977–983.

19. Pulendran B, Artis D. New paradigms in type 2 immunity. Science 2012;337:431–435.

20. Hirano T. Interleukin 6 and its receptor: 10 years later. Int Rev Immunol 1998;16:249–284.

21. Diehl S, Rincón M. The two faces of IL-6 on Th1/Th2 differentiation. Mol Immunol 2002;39(9):531–36.

22. Danso MO, van Drongelen V, Mulder A, van Esch J, Scott H, van Smeden J, et al. TNF-a and Th2 Cytokines Induce Atopic Dermatitis - Like Features on Epidermal Differentiation Proteins and Stratum Corneum Lipids in Human Skin Equivalents. Journal of Investigative Dermatology 2014;134:1941–1950.

23. Cameron DJ, Tong Z, Yang Z, Kaminoh J, Kamiyah S, Chen H, et al. Essential role of Elovl4 in very long chain fatty acid synthesis, skin permeability barrier function, and neonatal survival. Int J Biol Sci 2007;3:111–119.

24. van Smeden J, Janssens M, Kaye EC, Caspers PJ, Lavrijsen AP. The importance of free fatty acid chain length for the skin barrier function in atopic eczema patients. Exp Dermatol 2014;23:45–52.

25. Zhu Z, Oh M-H, Yu J, Liu YJ, Zheng T. The Role of TSLP in IL-13-induced atopic march. Sci Rep 2011;1:23.

26. Karin M, Ben-Neriah Y. Phosphorylation meets ubiquitination: The control of NF-kB activity. Annu Rev Immunol 2000;18:621–663.

27. Aupperle K, Bennett B, Han Z, Boyle D, Manning A, Firestein G. NF-k B regulation by I k B kinase-2 in rheumatoid arthritis synoviocytes. J Immunol 2001;166:2705–2711.

28. Monaco C, Andreakos E, Kiriakidis S, Mauri C, Bicknell C, Foxwell B, et al. Canonical pathway of nuclear factor k B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis. Proc Natl Acad Sci U S A 2004;101:5634–5639.

29. Zhang YL, Dong C. MAPkinase sinimmuneresponses. Cell Mol Immunol 2005;2:20–27.

30. Vestergaard C, Yoneyama H, Murai M, Nakamura K, Tamaki K, Terashima Y, et al. Overexpression of Th2-specific chemokines in NC/Nga mice exhibiting atopic dermatitis-like lesions. J Clin Invest 1999;104:1097–1105.

31. Sallusto F, Lenig D, Mackay CR, Lanzaveccchia A. Flexible programs of chemokine receptors expression on human polarized T helper 1 and 2 lymphocytes. J Exp Med 1998;187:875–883.

32. Vestergaard C, Bang K, Gesser B, Yoneyama H, Matsushima K, Larsen CG. A Th2 chemokine, TARC, prouduced by keratinocytes may recruit CLA+CCR4+ lymphocytes into lesional atopic dermatitis skin. J Invest Dermatiol 2000;115:640–646.

33. Kakinuma T, Nakamura K, Wakugawa M, Mitsui H, Tada Y, Saeki H, et al. Thymus and activation-regulated chemokine in atopic dermatitis: serum thymus and activation-regulated chemokine level is closely related with disease activity. J Allergy Clin Immunol 2001;107:535–541.

Article information Continued