NC/Nga 마우스에서의 아토피피부염 모델 개발 및 피부염 병변에 대한 다래 추출물(Actinidia Extract)의 효과

주영현 ( Young Hyun Joo ),원종현 ( Chong Hyun Won ),김지윤 ( Ji Yun Kim ),조광현 ( Kwang Hyun Cho ),민경업 ( Kyung Up Min ),김규한 ( Kyu Han Kim ) 대한피부과학회 2009 대한피부과학회지 Vol.47 No.10

Background: Atopic dermatitis is a chronic itchy, inflammatory skin disease that usually relapses. Although the etiology of atopic dermatitis remains unclear, it has been shown that both Th1 and Th2 cytokines play pathogenic roles in the generation of atopic dermatitis. DA-9102 is a fraction from the Actinidia species and DA-9102 displays immune modulating activity for allergy related disease. Objective: We have developed the atopic dermatitis model of NC/Nga mice using DNCB and we examined whether DA-9102 suppresses the development of atopic dermatitis-like skin lesions on NC/Nga mice. Methods: NC/Nga mice were challenged with DNCB during 5 weeks to develop atopic dermatitis-like skin lesions. Daily DA-9102 or cyclosporine A or HPMC (control) were then given orally. The efficacy of DA-9102 in NC/Nga mice was judged by measurement of the skin lesion severity (a modified SCORAD score), the serum IgE and IgG2a levels and the cytokine levels (IFN-γ and IL-4) from spleen cells cultured with ConA. Results: Atopic dermatitis-like lesions were developed on the NC/Nga mice by using topical DNCB. Oral administration of 100 mg/kg DA-9102 significantly suppressed the development of dermatitis, as was analyzed by a modified SCORAD score (p<0.01). The serum IgE level increased gradually with age, but treatment with DA-9102 suppressed the increment of the serum IgE level (p<0.01). The mean values of IFN-γ in the NC/Nga mice of the DA-9102 group were lower than those of the control mice group (p<0.05). The mean values of IL-4 were undetectable in all the experimental groups. The serum IgG2a level were not significantly different among all the experimental groups. Conclusion: We successfully developed an atopic dermatitis model in NC/Nga mice. Based on our in in vitro data, we suggest that DA-9102 can be useful for the treatment of atopic dermatitis. (Korean J Dermatol 2009;47(10):1105∼1112)

NC/Nga 마우스의 아토피 피부염에 미치는 카모마일, 라벤더, 샌달우드혼합오일 도포의 치유효과

신길란 ( Gil-ran Shin ),김양원 ( Yang-weon Kim ) 한국감성과학회 2016 감성과학 Vol.19 No.2

본 연구에서는 카모마일, 라벤더, 샌달우드를 혼합한 아로마 혼합오일(CLS)과 면역억제제(FK506)의 아토피 피부염 치료효과를 알아보기 위하여 혈청 중 IgE(Immunoglobulin E)와 IgG1(Immunoglobulin G1)의 수준의 감소효과, 피부 임상지수를 이용한 육안평가 등을 실시하였다. 그 결과 IgE 수준을 낮추는데는 CLS가 더 효과적이었고, IgG1수준을 감소시키는 데에는 FK506이 더 효과적인 것으로 나타났다. 육안평가 결과를 통하여 아토피 피부염을 가진마우스에 FK506과 CLS를 도포한 후 2주까지는 아토피 증상을 더 악화시켰으며 4주후부터는 증상완화가 현저하게나타남을 확인할 수 있었다. 특히 FK506의 효과가 두드러졌는데 이는 IgG1 수준감소와 관련이 있는 것으로 사료되며 이와 관련하여 보다 심도있는 실험 조사가 필요하겠다. 이상의 결과로부터 카모마일 저먼, 라벤더, 샌달우드의아로마 혼합오일이 아토피 피부염의 개선에 효능이 있음이 인정되었고 이와 같이 장기간 피부에 도포하여 치료하는경우 CLS가 아토피 치료의 한 방법으로서 가능성이 보임을 확인하였음에 본 연구의 의의가 있다. The effects of aroma mixed oil with Chamomile, Lavender and Sandalwood on atopic dermatitis in NC/Nga mice were examined. The NC/Nga mice were divided into BMAC group, FK 506 Oinment (Tacrolimus Hydrate) group, and CLS group to get curative power of CLS. The amount of total IgE and IgG1 was measured and the severity of atopic dermatitis was assessed by the scoring procedure in NC/Nga mice. Topically applied CLS significantly suppressed the level of serum IgE and IgG1 in NC/Nga mice and FK506, used as reference drugs for atopic dermatitis, also exhibited suppressive effects against level of IgE and IgG1. The level of IgE was lower in the CLS group than in the FK506 group while the serum IgG1 level in the FK506 group was lower than in the CLS group. The treatment with FK506 and CLS reduced the skin inflammation index, especially the severity degree of atopic dermatitis in the skin lesioned NC/Nga mice by naked eyes was improved by treatment of FK506 and CLS. The results suggest that treatment of CLS in NC/Nga mice with atopic dermatitis have an beneficial therapeutic effects on reducing the level of IgE and IgG1 and accelerating repair of skin lesion.

청열윤부탕(淸熱潤膚湯)이 DNFB로 유발된 알레르기 피부염에 미치는 효과

이경기,김진주,정희재,정승기,Lee, Kyung-Ki,Kim, Jin-Ju,Jung, Hee-Jae,Jung, Sung-Ki 대한한방내과학회 2008 大韓韓方內科學會誌 Vol.29 No.3

Objective : To examine the efficacy of CY, the improvement of atopy dermatitis(AD)-like lesions on the rostral back of the NC/Nga mice, which took CY orally and were treated with a chemical substance called DNFB, the inhibition of ear swelling, and the inhibition of inflammatory response of the ear tissue of the mice were observed and compared, and the inhibition of the serum IgE count and the IL-4 and IFN-${\gamma}$ count produced by CD4+ T cells of the lymph node were observed and compared. Materials and Methods : For measurement of ear thicknesses, 0.15% DNFB $25{\mu}l$ mixed with acetone/olive oil(3:1) was applied to the right ear and dorsal skin of the NC/Nga mice 5 times at an interval of 7 days. Ear thickness was measured every day over a five-week period after the first application of DNFB. The total serum IgE count was measured with ELISA by taking blood samples 24 hours after the fifth application of DNFB. To measure the IL-4 and IFN-${\gamma}$ count, the lymph node was cut off, and the CD4+ T cells were purified and stimulated to activate the T cells, and then the IL-4 and IFN-${\gamma}$ count was measured with ELISA. Results : Continuous oral administration of CY improved the AD-like skin lesions on the rostral back and inhibited the ear swelling in NC/Nga mice treated with DNFB and inhibited the inflammatory response in the NC/Nga mice treated with DNFB NC/Nga mice. Continuous oral administration of CY did not significantly inhibit the serum IgE count and failed to significantly reduce the IL-4 count generated after TCR stimulation in the CD4+ T cells of the NC/Nga mice treated with DNFB. Continuous oral administration of CY significantly reduced the IFN-${\gamma}$ count generated after TCR stimulation in the CD4+ T cells of the NC/Nga mice treated with DNFB.

<b><i>Costaria costata</i></b> Extract Suppresses Development of Atopic Dermatitis in chloro-2,4-dinitrobenzene-treated NC/Nga Mice

Kim, Ok-Kyung,Lee, Minhee,Kwon, Han Ol,Lee, Dasom,Park, Jeongjin,Kim, Eungpil,You, Yanghee,Lim, Young Tae,Jun, Woojin,Lee, Jeongmin S. Karger AG 2018 Skin pharmacology and physiology Vol.31 No.4

<P>We investigated the potential effects of<I> Costaria costata</I><B><I></I></B> (CC) on atopic dermatitis (AD) development in chloro-2,4-dinitrobenzene (DNCB)-treated NC/Nga mice. CC is a brown alga distributed across the seas of Korea, China, and Japan. A total of 40 mice were randomly assigned to 5 groups with 8 mice per group: untreated Balb/c mice, AD control (0.1% w/v DNCB-treated NC/Nga mice), positive control (i.e., DNCB-treated NC/Nga mice fed a dietary supplement of 66.6 mg/kg of body weight [b.w.] of CJLP133), DNCB-treated NC/Nga mice fed a dietary supplement of 100 mg/kg b.w. of CCE10 (CCE10 100), and DNCB-treated mice fed a dietary supplement of 300 mg/kg b.w. of CCE10 (CCE10 300) groups. The CCE10 100 and CCE10 300 treatment groups suppressed AD development including clinical and histopathological changes and a reduction in skin hydration induced by DNCB. In addition, Th2 cytokine production in primary splenocytes, serum IgE and histamine production, and mast cell infiltration into the skin were suppressed in the CCE10 300 mice compared to the CCE10 100 mice. Our finding demonstrated an inhibitory effect of CCE10 in AD development by means of improving the Th1/Th2 cytokine balance and anti-inflammatory effect in an in vivo model.</P>

가미청심연자탕(加味淸心蓮子場)이 NC/Nga mice의 아토피양(樣) 피부염에 미치는 영향

한재경,김윤희,윤지연,Han, Jae-Kyung,Kim, Yun-Hee,Yoon, Ji-Yeon 대한한방소아과학회 2007 대한한방소아과학회지 Vol.21 No.1

Objectives : The purpose of this study is to examine of the effect of Kami-chungsimyeunjatang(KCSYJT) medicine on the atopy eruption control. Methods : The expression of IgE, IL-4, IL-6, $TNF-{\alpha}$, IgG2b, IgM, IgG2a and IgG1 level in serum, and $IFN-{\gamma}$ production by KCSYJT were analyzed. CD3e+/CD69+, CD4+/CD25+, B220+/IgE+ and B220+/CD23+ positive cells by flow cytometry in splenocytes were assayed and the revelation of CD3e+/CD69+, CD4+/CD8+ and CD4+/CD25+ marker in PBMC, spleen and DLN were observed. The outturn of IL-4, eotaxin 2, CCR3, TARC mRNA in splenocytes werw observed. We also analyzed NC/Nga mice's ear, DLN and neck-back skin after biopy and dye by H&E, and toluidine staining (mast cells marker) method, measured about epidermis and dermis part in comparison with control group. Results : NC/Nga mice suffered from dermatitis very similar to human AD with IgE hyperproduction. Specially, result that measure IgE content in serum on 8 weeks, 12 weeks, 16 weeks decreased remarkably than control group. After experiment end, result that observe revelation CD3e+/CD69+, CD4+/CD8+ and CD4+/CD25+ marker in PBMC, spleen and DLN establishment observed recover as normal with political background. And decreased than result control group which measure IL-4, IL-6, $TNF-{\alpha}$, IgG2b, IgM, IgG2a, IgG1 level in serum, and $IFN-{\gamma}$ production secreted in Th1 cell displayed increase by KCSYJT medicines. Ear thickness decrease than control group in result that observe effect that get in ear of a NC/Nga mouse. Course inflammation immunocyte etc.. permeated of result that effect that KCSYJT medicines get to NC/Nga mouse's skin establishment analyzes ear, DLN and neck-back skin after biopy, and dye by H&E, and toluidine staining (mast cells marker) method decreased about epidermis. and inflammation of dermis part remarkably than control group. Immunohistochemical examination of the skin lesion showed decrease by KCSYJT medicines on numbers of mast cells (CCR3) and CD4+ T cells containing IL-4 necessary for IgE. Conclusions : Th1 cell and Th2 cell was observed to be shift by secretion amount of IL-4 and $IFN-{\gamma}$ by KCSYJT medicines. Therefore, the KCSYJT medicine turned out to be useful in allergy autoimmune disease.

현삼(玄蔘)이 NC/Nga mice에서 유발된 피부염에 미치는 영향

한재경,김윤희,여의주,Han, Jae-Kyung,Kim, Yun-Hee,Yeo, Eui-Ju 대한한방소아과학회 2007 대한한방소아과학회지 Vol.21 No.2

Objectives The purpose of this study is to examine of the effect of SPAR medicines on the atopy eruption control Methods This experiment is about the expression of IgE, IL-4, IL-6, IL_13, IgM, IgG2a, IgG2b, IgG1 level in serum, and $IFN-{\gamma}$ production by SPAR medicines. We assayed for $CD3e^+/CD69^+$, $CD044^+/CD19^+$ positive cells by flow cytometry in splenocytes and observed the revelation of $CD3e^+/CD69^+$, $CD4^+/CD8^+$, $CD44^+/CD19^+$ marker in PBMC, spleen and DLN. We also observed the outturn of IL-4, IL-5, CCR3, $IFN-{\gamma}$ in skin of a NC/Nga mice. We also analyzed NC/Nga mice's ear and neck-back skin after biopsy and dye by H&E staining method, measured about epidermis and dermis part in comparison with control group. Results SPAR medicines as treatment result to a NC/Nga mice, clinical skin severity score decreased remarkably than the ontrol group. Specially, experiment was results by measuring IgE and IL-6 content in serum 8 weeks, 10 weeks, 12 weeks, 16 weeks, 20 weeks respectively, and it was decreased remarkably than the control group. After experiment ended, the result that observed the revelation CD3e, CD4, CD8, CD19, CD69, CD11a marker in lymph node establishment were observed and that B/T rate becomes recover as normal with political background. In addition to that, the control group was decreased in the measured value of IL-4, IL-5, IL-13, IgM, IgG2a, IgG1's level in serum, and $IFN-{\gamma}$' production secreted in Th1 cell displayed increase by SPAR medicines. IL-4, IL-5, CCR3, and $IFN-{\gamma}$'s gene revelation amount displayed marked decrease than the control group in result that observe effect that get in skin of a NC/Nga dermatitis mouse. Moreover in culture supernatant which cultivate for 14 days after separate skin cell, IL-13 and IL-6 production, and $CD69^+/CD3e^+$, $CD44^+/CD19^+$ expression cell number was decreased than the control group's number. Course inflammation immunocyte permeated of result that effect that SPAR medicines get to NC/Nga mice's skin establishment analyzes ear and neck-back skin after biopsy, and dye by H&E method decreased about epidermis and inflammation of dermis part remarkably than the control group. Conclusions Th1 cell and Th2 cell observe to be shifted by secretion amount of IL-4 and $IFN-{\gamma}$ by SPAR medicines could know that SPAR medicines can be use for treatung allergy autoimmune disease.

Substance P restores normal skin architecture and reduces epidermal infiltration of sensory nerve fiber in TNCB-induced atopic dermatitis-like lesions in NC/Nga mice

Choi, Hyeongwon,Kim, Dong-jin,Nam, Seungwoo,Lim, Sunki,Hwang, Jae-Sung,Park, Ki Sook,Hong, Hyun Sook,Won, Younsun,Shin, Min Kyung,Chung, Eunkyung,Son, Youngsook Elsevier 2018 JOURNAL OF DERMATOLOGICAL SCIENCE Vol.89 No.3

<P><B>Abstract</B></P> <P><B>Background</B></P> <P>Atopic dermatitis (AD) is a chronic inflammatory skin disorder characterized by intense pruritus and eczematous lesion. Substance P (SP) is an 11-amino-acid endogenous neuropeptide that belongs to the tachykinin family and several reports recently have supported the anti-inflammatory and tissue repairing roles of SP.</P> <P><B>Objective</B></P> <P>In this study, we investigated whether SP can improve AD symptoms, especially the impaired skin barrier function, in 2, 4, 6-trinitrochlorobenzene (TNCB)-induced chronic dermatitis of NC/Nga mice or not.</P> <P><B>Method</B></P> <P>AD-like dermatitis was induced in NC/Nga mice by repeated sensitization with TNCB for 5 weeks. The experimental group designations and topical treatments were as follows: vehicle group (AD-VE); SP group (AD-SP); and SP with NK1R antagonist CP99994 (AD-SP-A) group. Histological analysis was performed to evaluate epidermal differentiation, dermal integrity, and epidermal nerve innervation in AD-like lesions. The skin barrier functions and pruritus of NC/Nga mice were evaluated by measuring transepidermal water loss (TEWL) and scratching behavior, respectively.</P> <P><B>Result</B></P> <P>Topical SP treatment resulted in significant down-regulation of Ki67 and the abnormal-type keratins (K) K6, K16, and K17, restoration of filaggrin and claudin-1, marked reduction of TEWL, and restoration of basement membrane and dermal collagen deposition, even under continuous sensitization of low dose TNCB. In addition, SP significantly reduced innervation of itch-evoking nerve fibers, gelatinase activity and nerve growth factor (NGF) expression in the epidermis but upregulated semaphorin-3A (Sema3A) expression in the epidermis, along with reduced scratching behavior in TNCB-treated NC/Nga mice. All of these effects were completely reversed by co-treatment with the NK1R antagonist CP99994. In cultured human keratinocytes, SP treatment reduced expression of TGF-α, but upregulated TGF-β and Sema3A.</P> <P><B>Conclusion</B></P> <P>Topically administered SP can restore normal skin barrier function, reduce epidermal infiltration of itch-evoking nerve fibers in the AD-like skin lesions, and alleviate scratching behavior. Thus, SP may be proposed as a potential medication for chronic dermatitis and AD.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Topical Substance P treatment reduced AD-like symptoms in TNCB-induced NC/Nga mice. </LI> <LI> Topical Substance P treatment reduced epidermal acanthosis and expressed normal keratin profiles in AD-like mice. </LI> <LI> Topical Substance P treatment regulated both small nerve attraction and repulsion factors in AD-like skin. </LI> <LI> Topical Substance P treatment recovered skin barrier function and reduced pruritus in AD-like mice. </LI> </UL> </P>

라벤더, 레몬, 유칼립투스 혼합 에센셜오일이 아토피 피부염 동물 모델의 Th2 관련인자에 미치는 영향

김현아,윤미영,송향희,정광조,유화승,Kim, Hyeon-Ah,Yun, Mi-Young,Song, Hyang-Hee,Cheong, Kwang-Jo,Yoo, Hwa-Seung 대한약침학회 2010 Journal of pharmacopuncture Vol.13 No.1

Purpose : To investigate the effects of the lavender, lemon and eucalyptus oil mixture on the atopy dermatitis skin lesions induced on NC/Nga Mice by dinitrochlorobenzene (DNCB). Material and Method : For this purpose, we fabricated the oil mixture blending three essential oils (lavender, lemon, eucalyptus : ELL) with one carrier oil (jojoba) and apply it on the atopic dermatitis skin lesions of NC/Nga Mice. Atopic dermatitis in NC/Nga Mice was induced by DNCB treatment on the dorsal skin of mice for 8 weeks. The mixture of ratio of each essential oil drop was 1 (eucalyptus) : 2 (lemon) : 2 (lavender) and this mixture was blended with jojoba oil 50ml (0.025%). The ELL-ointment was supplied for 8 weeks. We evaluated the effects of ELL on cell viability of mouse lung fibroblast, clinical skin features and severity, the level of serum Immunoglobulin (Ig) E & Ig G1, Interleukin (IL)-4, IL-13 and Interferon (IFN)-$\gamma$. Results : ELL showed safety on the cell viability of mouse lung fibroblast compared with control group. The cell viability was measured by SRB method. The effects of ELL on clinical skin features and severity in DNCB-induced dermatitis model of NC/Nga mice was significant compared with control group. EEL also showed significant effects on clinical symptom score compared with control group. Serum IgE & IgG1 level and development of atopy dermatitis skin lesions were evaluated. Serum IgE & IgG1 production was significantly down-regulated in EEL group compared with control group. ELL also down-regulated the levels of IL-4 and IL-13, and up-regulated the level of IFN-$\gamma$ compared with control group significantly. Conclusion : ELL was effective on atopy dermatitis by modulating Th2 related factors.

이중탕(理中湯)이 Mite Antigen으로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향

서희연,한재경,김윤희,Seo, Hui-Yeon,Han, Jae-Kyung,Kim, Yun-Hee 대한한방소아과학회 2011 대한한방소아과학회지 Vol.25 No.1

Objectives: The purpose of this study is to investigate the effects of Yijungtang(YJT) on atopic dermatitis in an in-vitro and in-vivo experiment using a RBL-2H3 mast cells and a NC/Nga atopic dermatitis mouse. Methods: In-vitro experiment, IL-4, IL-13 mRNA expression were evaluated by a real-time PCR, IL-4, IL-13 production by ELISA and transcription factor as GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-kB by western blotting. In-vivo experiment, clinical skin score we evaluated by, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, cytokine level, total number of cell, Immunohistochemical staining and Histological features of auxiliary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results: YJT decreased IL-4, IL-13 mRNA expression, IL-4, IL-13 production and prominently decreased the expression of mast cell specific transcription factors including GATA-2, NF-AT2, c-Fos and NF-kB. YJT oral administration reduced the levels of skin severity scores. It also decreased the level of inflammatory cytokines such as IL-5, IL-13, histamine and IgE in the serum. It elevated IFN-gamma level in the spleenocyte culture supernatant but decreased. $CD3e^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $CD11b^+Gr-1^+$, $CCR3^+$ in the PBMCs, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $B220^+CD23^+$ in the ALN, $CD4^+$, $CD3e^+CD69^+$ in the ALN and $CD4^+$, $CD11b^+Gr-1^+$ in the dorsal skin. Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mice were much improved by YJT oral administration. Conclusions: The anti-allergic activities of YJT may be mediated by down-regulation of Th2 cytokines, such as IL-4 and IL-13, through the regulation GATA-2, NF-AT2 and NF-kB transcription factors in mast cells. YJT would be regulate molecular mediators and immune cells which are functionally associated with atopic dermatitis induced in NC/Nga mice, and may play an important role in recovering AD symptoms.

구풍제습탕(驅風除濕湯)이 DNCB로 유도된 NC/Nga mice의 아토피 피부염에 미치는 영향

윤재은,김윤희,한재경,김윤희,Yoon, Jae-Eun,Kim, Yun-Hee,Han, Jae-Kyung,Kim, Yun-Hee 대한한방소아과학회 2008 대한한방소아과학회지 Vol.22 No.3

Objectives : The purpose of this study is to investigate the effect of Gupoongjeseuptang (GPJST) on atopic dermatitis by in vivo experiment using NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the atopic dermatitis of human. Methods : To investigate the effect of GPJST on atopic dermatifis, we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells (PBMCs), splenocytes, draining lymph node (DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis-like skin NC/Nga mouse in vivo. Results : In vivo, clinical skin severity score were significantly lower in GPJST group than control group. IgE, IL-6, $TNF-{\alpha}$, IgG1, IgM, IgG2a and IgG2b levels in serum decreased remarkably in GPJST group than control group. Also, total absolute number of $CD3^+CD69^+$, and $CCR3^+$ cells recovered as normal in PBMCs and $CD3^+$, $CD3^+CD69^+$ decreased significantly compared with control group in isolated DLN from NC/Nga mouse and total absolute number of $CD11b^+Gr-1^+$, $CCR3^+CD3^+$ in dorsal skin of NC/Nga mouse decreased by GPJST. We analyzed ear and neck-back skin after biopsy and dyeing by hematoxyline/eosin (H&E) and toluidine staining (mast cells marker) and obtained results that GPJST are very effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell (CD4, $CCR3^+$) infiltration). Conclusions : This study demonstrates immunological activity of GPJST on atopic dermatitis-like model mice.