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JKM > Volume 36(1); 2015 > Article
Lee, Choi, and Yang: Effects of topical application of realgar on pruritus and inflammation of atopic dermatitis

Abstract

Objectives:

Realgar has been frequently used for skin disorders in history of herbal medicine. However, the efficacy of realgar has not been examined in atopic dermatitis (AD). In this study, the effects of realgar on AD were investigated, especially on pruritus and inflammation.

Methods:

AD lesions were induced in the shaved backs of BALB/c mice through repeated application of DNCB. The mice were treated for 11 days with 1% realgar (100 μL/day). Histological changes in skin thickness were observed. The anti-pruritic effects of realgar were evaluated by the change in numbers of scratching behavior of mice and expression of substance P. The expressions of cytokines IL-4 and IL-6 were measured. Also, anti-inflammatory effects of realgar were examined on expressions of NF-κB, phospho-I κB α and mitogen-activated protein kinases (MAPKs).

Results:

Realgar decreased skin thickness (both dermal and epidermal) 38% and 17% respectively, compared to positive control, DNCB group. The scratching behavior of mice was reduced by 42% and expression of substance P was significantly less. Cytokines IL-4 and IL-6 were significantly reduced by 52.6% and 77.6%, respectively. The expressions of NF-κB, phospho-I κBα and MAPKs (phospho-ERK1/2, -p38 and -JNK) were significantly suppressed with marked effects on phospho-ERK1/2.

Conclusions:

The collective results suggest that realgar shows anti-pruritic and anti-inflammatory effects on AD. And realgar might be a potential therapeutic candidate for treatment of atopic dermatitis.

Fig. 1.
Experimental schedule. ▲: The shaved dorsal skin was applied with 100 µL of 1% DNCB solution (in acetone: olive oil=4:1) for sensitization, ●: The realgar group were treated 1% realgar (5 mg/day) and the DEX group treated 100 µL dexamethasone (10 uM, dissolved in PBS), △: 100 µL of 0.5 % DNCB solution were applied on the dorsal skin.
jkm-36-1-9f1.tif
Fig. 2.
Change in epidermis and dermis upon application of realgar. (A) The thickness of epidermis, (B) The thickness of dermis. Hematoxylin-eosin staining showed a decrease thickness of epidermis and dermis in the realgar-treated group and DEX group. Results shown are representative of five observations. Data are presented as means ± SD and analyzed with ANOVA. ###P < 0.001, versus the NOR group. ***P < 0.001, versus the DNCB group. Magnification : X 100.
jkm-36-1-9f2.tif
Fig. 3.
Changes in scratching behavior upon application of realgar. Scratching behavior of mouse was checked after 1 h had passed following the last application of DNCB (on 18 day). Each mouse of all group were videotaped for 20 min to count the number of scratching behavior. Data are presented as means ± SD (n = 5) and analyzed with ANOVA. ###P < 0.001, versus the NOR group. **P < 0.05, versus the DNCB group.
jkm-36-1-9f3.tif
Fig. 4.
Changes in substance P expression upon application of realgar. The expression of substance P was confirmed by Immunohistochemistry staining. Skin specimens were excised 12 h after the final DNCB challenge. The arrow indicates the expression of substance P. Magnification : X 200.
jkm-36-1-9f4.tif
Fig. 5.
Changes in cytokine levels upon application of realgar. The cytokine levels of IL-4 and IL-6 were measured in the dorsal skin using mouse ELISA kits. Data are presented as means ± SD (n = 5) and analyzed with ANOVA.###P < 0.001, versus the NOR group. ***P < 0.001, versus the DNCB group.
jkm-36-1-9f5.tif
Fig. 6.
Changes in expressions of NF-κB, phospho-IκBα upon application of realgar. The effects of realgar on the expressions of NF-κB and phospho-IκBα was assessed using western blot analysis. Results shown are representative of five observations. Data are presented as means ± SD and analyzed with ANOVA. ###P < 0.001, versus the NOR group. ***P < 0.001, **P < 0.01, versus the DNCB group.
jkm-36-1-9f6.tif
Fig. 7.
Changes in MAPKs upon application of realgar. (A) phospho-ERK1/2, (B) phospho-p38 and (C) phospho-JNK. Total protein of the dorsal skin was analyzed by western blotting. Results shown are representative of five observations. Data are presented as means ± SD and analyzed with ANOVA. ###P < 0.001, versus the NOR group. ***P < 0.001, **P < 0.01, versus the DNCB group.
jkm-36-1-9f7.tif
Fig. 8.
Changes in liver tissues upon application of realgar. Results shown are representative of five observations. Magnification: X 200.
jkm-36-1-9f8.tif

참고문헌

1.. Bieber T. Atopic dermatitis. N Engl J Med. 2008; 358:1483–94.
crossref

2.. Akdis CA, Akdis M, Bieber T, Bindslev-Jensen C, Boguniewicz M, Eigenmann P, et al. Diagnosis and treatment of atopic dermatitis in children and adults: European Academy of Allergology and Clinical Immunology/American Academy of Allergy, Asthma and Immunology/PRACTALL Consensus Report. J Allergy Clin Immunol. 2006; 118:1. 152–169.
crossref

3.. DaVeiga SP. Epidemiology of atopic dermatitis: a review. Allergy Asthma Proc. 2012; 33:3. 227–234.
crossref

4.. Ständer S, Luger TA. Itch in atopic dermatitis–pathophysiology and treatment. Acta Dermatovenerol Croat. 2010; 18:4. 289–296.


5.. Klein PA, Clark RAF. An Evidence-Based review of the efficacy of antihistamines in relieving pruritus in atopic dermatitis. Arch Dermatol. 1999; 135:12. 1522–1525.
crossref

6.. Sher L, Chang J, Patel B, Balkrishnan R, Fleischer AB. Relieving the Pruritus of Atopic Dermatitis: A Meta-analysis. Value Health. 2012; 15:4. A249–A250.
crossref

7.. Sohn KH, Lee JG, Jee SY. A philological study on demonstration of atopic dermatitis. J East-West Med. 2009; 34:4. 15–24.


8.. 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.
crossref

9.. Park BK, Chang GT, Kim JH. A study of external treatments for the atopic dermatitis. J Korean Orient Pediatr. 2003; 17:1. 71–86.


10.. Wu YL. Bonchojongsin. First edition. Seoul: Haeng Lim Publishers Co;1982. p. 143


11.. Heo J. Newly translated Donguibogam. Second edition. Seoul: Bubin Publishers Co;2009. p. 797–810.


12.. Lee KS, Jeong ES, Heo SH, Seo JH, Jeong DG, Choi YK. A Novel Model for Human Atopic Dermatitis: Application of Repeated DNCB Patch in BALB/c Mice, in Comparison with NC/Nga Mice. Lab Anim. 2010; 26:1. 95–102.
crossref

13.. Kabashima K. New concept of the pathogenesis of atopic dermatitis: Interplay among the barrier, allergy, and pruritus as a trinity. J Invest Dermatol. 2013; 70:3–11.
crossref

14.. Cork MJ, Danby SG, Vasilopoulos Y, Hadgraft J, Lane ME, Moustafa M, et al. Epidermal barrier dysfunction in atopic dermatitis. J Invest Dermatol. 2009; 129:8. 1892–1908.
crossref

15.. Ye YM, Kim BE, Shin YS, Park HS, Leung DYM. Overexpression of Epidermal Filaggrin in Patients with Chronic Idiopathic Urticaria Correlates with Urticaria Severity. J Allergy Clin Immunol. 2013; 131:2. AB56
crossref

16.. Koblenzer CS. Itching and the atopic skin. J Allergy Clin Immunol. 1999; 104:3. S109–S113.
crossref

17.. Gupta MA, Gupta AK. Depression modulates pruritus perception. A study of pruritus in psoriasis, atopic dermatitis and chronic idiopathic urticaria. Ann N Y Acad Sci. 1999; 885:394–395.
crossref

18.. Shim WH, Song CH, Park HJ, Kim HS, Chin HW, Kim SH, et al. Clinical characteristics of itch in patients with adult and childhood atopic dermatitis. Korean J Dermatol. 2011; 49:4. 318–327.


19.. Park SW, Yan YP, Satriotomo I, Vemuganti R, Dempsey RJ. Substance P is a promoter of adult neural progenitor cell proliferation under normal and ischemic conditions. J. Neurosurg. 2007; 107:3. 593–599.
crossref

20.. Donkin JJ, Turner RJ, Hassan I, Vink R. Substance P in traumatic brain injury. Prog Brain Res. 2007; 161:97–109.
crossref

21.. Ebner K, Singewald N. The role of substance P in stress and anxiety responses. Amino Acids. 2006; 31:3. 251–272.
crossref

22.. Yamaoka J, Kawana S. Rapid changes in substance P signaling and neutral endopeptidase induced by skin-scratching stimulation in mice. J Dermatol Sci. 2007; 48:2. 123–132.
crossref

23.. Hon KLE, Lam MCA, Wong KY, Leung TF, Ng PC. Pathophysiology of Nocturnal Scratching in Childhood Atopic Dermatitis: The Role of Brain-Derived Neurotrophic Factor and Substance P. Br J Dermatol. 2007; 157:5. 922–925.
crossref

24.. Toyoda M, Nakamura M, Makino T, Hino T, Kagoura M, Morohashi M. Nerve growth factor and substance P are useful plasma markers of disease activity in atopic dermatitis. Br J Dermatol. 2002; 147:1. 71–79.
crossref

25.. Mosmann TR, Coffmann RL. Heterogeneity of cytokine secretion patterns and functions of helper T cells. Adv Immunol. 1989; 46:111–139.
crossref

26.. Ogawa H, Mukai K, Kawano Y, Minegishi Y, Karasuyama H. Th2-inducing cytokines IL-4 and IL-33 synergistically elicit the expression of transmembrane TNF-α on macrophages through the autocrine action of IL-6. Biochem Biophys Res Commun. 2012; 420:1. 114–118.
crossref

27.. Werner S, Krieg T, Smola H. Keratinocyte-fibroblast interactions in wound healing. J Invest Dermatol. 2007; 127:5. 998–1008.
crossref

28.. Perkins ND. Integrating cell-signalling pathways with NF-kappaB and IKK function. Nat Rev Mol Cell Biol. 2007; 8:1. 49–62.
crossref

29.. Bonizzi G, Karin M. The two NF-κ B activation pathways and their role in innate and adaptive immunity. Trends Immunol. 2004; 25:6. 280–288.
crossref

30.. Monaco C, Andreakos E, Kiriakidis S, Mauri C, Bicknell C, Foxwell B, et al. Canonical pathway of nuclear factor kappa B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis. Proc Natl Acad Sci USA. 2004; 101:15. 5634–5639.
crossref

31.. Arbabi S, Maier RV. Mitogen-activated protein kinases. Crit Care Med. 2002; 30:1. S74–S79.
crossref

32.. Garrington TP, Johnson GL. Organization and regulation of mitogen-activated protein kinase signaling pathways. Curr Opin Cell Biol. 1999; 11:2. 211–218.
crossref

33.. Hong ZY, Xiao M, Yang Y, Han ZQ, Cao Y, Li CR, et al. Arsenic disulfide synergizes with the phosphoinositide 3-kinase inhibitor PI-103 to eradicate acute myeloid leukemia stem cells by inducing differentiation. Carcinogenesis. 2011; 32:10. 1550–1558.
crossref

34.. Liu J, Lu Y, Wu Q, Goyer RA, Waalkes MP. Mineral arsenicals in traditional medicines: orpiment, realgar, and arsenolite. J Pharmacol Exp Ther. 2008; 326:363–368.
crossref

35.. Wester RC, Hui X, Barbadillo S, Maibach HI, Lowney YW, Schoof RA, et al. In Vivo Percutaneous Absorption of Arsenic from Water and CCA-Treated Wood Residue. Toxicol Sci. 2004; 79:2. 287–295.
crossref

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