Anti-atopic dermatitis effects of  Rafinesque via regulation of immune response and nerve growth factor

Seon A Jung; You Yeon Choi; Woong Mo Yang; Jung, Seon A; Choi, You Yeon; Yang, Woong Mo

doi:10.13048/jkm.16002

JKM > Volume 37(1); 2016 > Article

Jung, Choi, and Yang: Anti-atopic dermatitis effects of Poncirus trifoliata Rafinesque via regulation of immune response and nerve growth factor

Original Article

The Journal of Korean Oriental Medicine 2016; 37(1): 10-20.

Published online: March 31, 2016

DOI: https://doi.org/10.13048/jkm.16002

Anti-atopic dermatitis effects of Poncirus trifoliata Rafinesque via regulation of immune response and nerve growth factor

Seon A Jung, You Yeon Choi, Woong Mo Yang

Department of Convergence Korean Medical Science, College of Korean Medicine, Kyung Hee University

Correspondence to: 양웅모(Woong Mo Yang), 서울시 동대문구 회기동 1번지 경희대학교 한의과대학, Tel: +82-2-961-2209, E-mail: wmyang@khu.ac.kr

Received November 25, 2015 Revised March 14, 2016 Accepted March 17, 2016

Abstract

Objectives:

Poncirus trifoliata Rafinesque has been known to have anti-allergic effects in skin diseases. However, anti-atopic dermatitis effects of P. trifoliata Rafinesque have not been studied yet in skin diseases. The present study evaluated the anti-atopic dermatitis effects of P. trifoliata Rafinesque (PTR) using external treatments on AD.

Methods:

AD lesions were induced by the repeated application of 2, 4-Dinitrochlorobenzene (DNCB) on the shaved back of BALB/c mice. 100 μℓ of PTR extracts was applied to the AD lesions for 11 days. Histological assessments, mast cells count and serum levels of IgE were analyzed. The anti-pruritic effects of PTR were examined by the change of scratching frequency and nerve growth factor (NGF) expression. In addition, the anti-inflammatory effects of PTR were examined by the expressions of Th2/Th1 cytokines and pro-inflammatory in dorsal skin.

Results:

Histopathological findings showed that topical application of PTR decreased the thickness of dermal and epidermal skin compared with the DNCB group. PTR also notably decreased the mast cells count and serum IgE. The scratching behavior of mice and expression of NGF were significantly reduced. In addition, PTR group significantly suppressed the IL-4, IL-6, IFN-γ and TNF-α cytokines compared to the DNCB group.

Conclusions:

These results indicated that P. trifoliata Rafinesque possess anti-pruritus and anti-atopic dermatitis properties. Therefore, P. trifoliata Rafinesque might be used for treatment of pruritus and atopic dermatitis.

Keywords: Ponciri fructus, atopic dermatitis, anti-pruritus, anti-inflammation

Fig. 1.

Experimental scheme for atopic dermatitis induction and PTR application.

Fig. 2.

Effects of PTR on skin thickness in DNCB-induced mice. (A) The thickness of epidermis, (B) the thickness of dermis. Hematoxylin-eosin staining showed a decrease thickness of epidermis and dermis in the PTR-treated 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.

Fig. 3.

Effects of PTR on the number of mast cells in DNCB-induced mice. Toluidine blue staining showed a decrease the number of mast cells in the RTR 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, ^** P < 0.01 versus the DNCB group. Magnification: X 100.

Fig. 4.

Effects of PTR on serum IgE level in DNCB-induced mice. Data are presented as means ± SD and analyzed with ANOVA ^### P < 0.001, versus the NOR group. ^*** P < 0.001, versus the DNCB group.

Fig. 5.

Effects of PTR on scratching behavior in DNCB-induced mice. 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. N.S: non-significant.

Fig. 6.

Effects of PTR on NGF expression in DNCB-induced mice. The expression of NGF was confirmed by western blotting assay. ^### P < 0.001, versus the NOR group. ^## P < 0.01, versus the DNCB group. N.S: non-significant.

Fig. 7.

Effects of PTR on levels of cytokine in DNCB-induced mice. The cytokine levels of IL-4, −6, IFN-γ and TNF-α 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 ^* P < 0.05, versus the DNCB group.

참고문헌

1.. Min DL, Park EJ, Kang KH. Review of Clinical and Experimental Studies on External Application Treatment for Atopic Dermatitis in the Korean Literature. J Korean Orient Pediatr. 2013; 27:1. 36–49.

2.. Park Y. Status of clinical practice on diagnosis and management of atopic dermatitis in Korea: a questionnaire survey of physicians. Allergy Asthma Respir Dis. 2013; 1:3. 257–265.

3.. Lee YJ, Kang HY, Jeong BJ, Kim KE, Lee KY, Lee SY. Significance of atopic index in children with atopic dermatitis. Allergy Asthma Respir Dis. 1994; 4:2. 11–24.

4.. Choi HJ, Sim BY, Miyamoto T. Effects of Lobophytum crassum extract (MC-1) on Various Immunological Factors Related to Pathogenesis of Atopic Dermatitis in Dermatophagoides Pteronyssinus Treated NC/Nga mice. Korea J Herbol. 2015; 30:1. 95–101.

5.. Yun HJ, Ko WS. Clinical Study of Atopic Dermatitis; the Classification of Oriental Medical Clinical type and Treatment. J Korean Oriental Med. 2001; 22:2. 10–21.

6.. Lee SH, Yun YG. Study for Treatment of Atopic Dermatitis in Oriental Medical Prescription. J Korean Med. 2006; 15:1. 56–69.

7.. Shin SH, Kim MB, Byun SM, Lee HS, Park SG, Ko WS, et al. A Review on External Treatment for Atopic Dermatitis in Korean Journals. J Korean Orient Med Ophthalmol Otolaryngol Dermato. 2008; 21:2. 80–93.

8.. Katayama I, Kohno Y, Akiyama K, Ikezawa Z, Kondo N, Tamaki K, et al. Japanese Society of Allergology. Japanese Guideline for Atopic Dermatitis. Allergol Int. 2011; 60:205–220.

9.. Boneberger S, Rupec RA, Ruzicka T. Complementary therapy for atopic dermatitis and other allergic skin diseases: facts and controversies. Clin Dermatol. 2010; 28:1. 57–61.

10.. Cha KB, Kim YS, Seol IC. Literatual study on Atopic dermatitis. Daejeon J Institute Orient Med. 2005; 14:2. 113–126.

11.. Kim EH, Lee BW, Kim EH. A Literal Study of the Main Symptoms of Atopic Dermatitis. J Korean Med Classic. 2009; 22:1. 207–228.

12.. College of korean medicine co association editorial board. Textbook of herbology. Seoul: Younglimsa;2011. p. 392

13.. Aeom YD, Kim DH, Jeong JG, Shin MK, Song HJ. The Comparative Study of Fructus Immaturus Ponciri and Fructus Ponciri Effect on Allergic Re action. J Korean Orient Med. 2001; 22:4. 10–21.

14.. Kang BS. Korean medicine clinical allergy. Seoul: Seongbosa;1988. p. 57–59.

15.. Hwang KH, Lee EJ, Song BK, Kim HK. Mechanism of Inhibitory Action of Anaphylaxis by Aqueous Extract of Poncirus trifoliata. J Korean Orient Med. 1997; 18:1. 316–325.

16.. Williams HC. Clinical practice. Atopic dermatitis. N Engl J Med. 2005; 352:22. 2314–2324.

17.. Cho JB, Lee JH, Suh JM, Yu JS, Lee HS, Park EA, et al. Change in Quality of Life according to the Change in Atopic Dermatitis Severity. Allergy Asthma Respir Dis. 2012; 22:1. 86–99.

18.. Carstens E, Akiyama T. Itch: Mechanisms and Treatment. Boca Raton (FL): CRC Press/Taylor & francis;2014. Ebook-Chapter 3.

19.. Lee YJ, Kang HY, Jeng BJ, Kim GE, Lee GY, Lee SY. Significance of atopic index in children with atopic dermatitis. Allergy Asthma Respir Dis. 1994; 4:2. 11–24.

20.. Leung DY, Bieber T. Atopic dermatitis. Lancet Diabetes Endocrinol. 2003; 361:151–160.

21.. Hong SJ. Review Articles: Mast Cells and Allergic Diseases. Allergy Asthma Respir Dis. 2000; 10:3. 207–217.

22.. Hwang SY, Lee SH, Lee CW, Kim JH, Jang SI, Kim AN, et al. Anti-inflammatory and Anti-Pruritonic Effects of WSY-1075 composited with Medicinal Plants on the Activated Rat Peritoneal Mast cells and Mouse Pruritus. Korea J Herbol. 2013; 28:4. 93–100.

23.. Metz M, Grundmann S, Ständer S. Pruritus: an overview of current concepts. Vet Dermatol. 2011; 22:2. 121–131.

24.. Yamaguchi J, Aihara M, Kobayashi Y, Kambara T, Ikezawa Z. Quantitative analysis of nerve growth factor (NGF) in the atopic dermatitis and psoriasis horny layer and effect of treatment on NGF in atopic dermatitis. J Dermatol Sci. 2009; 53:1. 48–54.

25.. Pincelli C, Marconi A. Autocrine nerve growth factor in human keratinocytes. J Dermatol Sci. 2000; 22:2. 71–79.

26.. Lim JH, Kang H, Park YM, Cho SH, Lee SY. Effect of substance P on Production of interleukin-12 and interferon-gamma from peripheral blood mononuclear cells of chronic severe atopic dermatitis patients. J Asthma. 2001; 21:4. 636–646.

27.. Lee IJ. Epidermal cytokine profiles in allergic and irritant contact dermatitis. Yonsei Univ Grad School, Doctoral dissertation.2002. 28

28.. Yu JS, Kim JU, Lee CH, Lee SR, Yook TH. The Effects of Lonicerae Flos, Forsythiae Fluctus and Hwangryunhaedok Decoction Pharmacopuncture on Atopic Dermatitis in NC/Nga Mice. J Korean Acupunct Moxibust Soc. 2015; 32:4. 119–131.

29.. Yu HS, Gim S, Song H, Ji J, Bal JW, Kim D. A Study on the Immune Modulation of Chunghwatang (CHT) in Atopic Dermatitis Animal Models. Daejeon J Institute Orient Med. 2012; 21:1. 53–64.