• The Journal of Pediatrics of Korean Medicine
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• v.30 no.4
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• pp.77-86
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• 2016

Objectives Hataedock (HTD) is an oral Korean herbal medical oral treatment that removes fetal toxic heat and meconium from new born babies. The purpose of this study is to evaluate whether Hataedock treatment of Duchi extracts has anti-inflammation effects in atopic dermatitis-like skin lesions in House Dust Mite-Induced NC/Nga Mice. Methods The mice were divided into 3 groups (n=10 per group) as follows: the control group (Ctrl group), AD-induced group (AE group), AD-induced with HTD treatment group (DT group). 3-week-old NC/Nga mice were introduced to Hataedock treatment, made of Duchi extract. After 4 weeks, House Dust Mite-Induced application was used six times per week for 3 weeks to induce the first atopic dermatitis, and second AD in 7 weeks after. To examine skin injuries and anti-inflammatory effect, PKC, MMP-9, iNOS immunohistochemistry were used. Results The alleviate effect of the skin damage and angiogenesis was observed in DT group. The damage of stratum corneum, hyperplasia, edema, infiltration of lymphocytes and distribution of capillary were decreased in DT group. Also, the study results suggested that Hataedock treatment made of Duchi extracts in DT group remarkably decreased skin damages by 51% (p < 0.001), as well as PKC by 91%, MMP-9 by 48% (p < 0.001), iNOS by 51% (p < 0.001). Conclusions Based on the study results, we observed that Hataedock treatment of Duchi extracts alleviates AD by diminishing various inflammatory cytokines, initial steps of AD development, in the skin lesions. Potential applications for prevention and treatment of atopic dermatitis are expected.

• The Journal of Pediatrics of Korean Medicine
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• v.23 no.2
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• pp.29-50
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• 2009

Objectives : The purpose of this study is to investigate effectiveness of KamiCheongsimyeonjatang(KCSYJT) medicines to suppress atopic dermatitis in mouse model experimentally. Methods : First, in vitro, we isolated B cells from 18 weeks of atopicdermatitis-like skin NC/Nga mouse. Then we analyzed FACS(Fluorescence Activated Cell Sorter) by intracellular staining of IFN-$\gamma$, GATA-3+ analyzed cytokines by using real-time PCR. Secondly, in vivo, after administration of KCSYJT to atopic dermatitis NC/Nga mouse at 12 weeks of age, we analyzed serum IgE and the change of activated cell in PBMCs(Peripheral Blood Mononuclear Cells). Results : In vitro, KCSYJT medicines supressed IL-1$\beta$, IL-6, TNF-$\alpha$, and TGF-$\beta$ mRNA and increased IL-10 mRNA in B cells. Also, KCSYJT medicines decreased the levels of GATA-3$^+$CD4$^+$ and increased the levels of IFN-$\gamma^+$CD4$^+$T Cell. In vivo, serum IgE levels dreased in KCSYJT group than control group and In PBMCs, the activated cell percentage of granulocytes, CD3+, CD3+/CD4+, B220+/CD23+, and CCR3+ decreased and CD19+, CD3+/CD8+ increased in KCSYJT group than control group. Conclusions : This study demonstrates immunological activity of KCSYJT on atopic dermatitis-like model mice.

• The Journal of Pediatrics of Korean Medicine
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• v.23 no.1
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• pp.1-22
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• 2009

Objectives The purpose of this study is to investigate the effect of Li Zhong Tang(LZH-T) on atopic dermatitis by using NC/Nga atopic dermatitis mouse. Methods First, in vitro, we isolated B cells from NC/Nga mouse which induced atopic dermatitis-like skin for 18 weeks. We analyzed FACS by intracellular staining of $IFN-{\gamma}$, GATA-3+ and also analyzed cytokines by using real-time PCR. Secondly, in vivo, after administration of LZH-T to the 12 weeks old mouse with atopic dermatitis. We analyzed serum IgE, $IFN-{\gamma}$ level and observed the changes of activated cell. Results In vitro, LZH-T decreased the levels of CD4+/$IFN-{\gamma}$ and increased the levels of CD4+/GATA3+. In vivo, serum IgE levels were decreased and $IFN-{\gamma}$ levels were increased in LZH-T group compared to the control group. In PBMCs, the percentage of activated cell - granulocytes, CD3+, CD3+CD4+, B220+CD23+, and CCR3+ were decreased and CD19+, CD3+CD8+ were increased in LZH-T group compared to the control group. Conclusions This study demonstrates immunological activity of GPJST on atopic dermatitis-like model mice.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.2 s.33
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• pp.142-150
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• 2007

Objectives : This study was carried out to investigate the latest study tendency in mouse model of skin pruritus published since 2005 and to arrange various experimental methods. Methods : We examined the journal(such as Experimental dermatology and British journal of dermatology) and in Pubmed since 2005, regarded pruritus and scratching behavior model as key words. Results and Conclusion : 1. BALB/c, hairless, NC/Nga and ICR mice were the most used in scratching behavior model. According to scratching-induced agents. we need to select experimental mice. 2. There are various methods inducing skin pruritus; by cohabitation with NC/Nga mice having severe skin lesions, by injection intradermally(or subcutaneously) with agent inducing inflammation, by inducing contact dermatitis with TNCB, TNFB, destruction of skin barrier and by transgenic mice. 3. Injection intradermally(or subcutaneously) with agent inducing inflammation is the most used out of methods inducing skin pruritus. Compound 48/80, histamine, substance P and others(chloroquine, serotonin, carrageenin, TPA etc.) were included in agents inducing inflammation and pruritus in skin pruritus model. 4. Video camera, SCLABA system, MicroAct and acoustic evaluation system were included in evaluation methods of scratching behavior mouse model.

• 한국약용작물학회:학술대회논문집
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• 2008.11a
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• pp.417-418
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• 2008

• Toxicological Research
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• v.22 no.4
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• pp.357-364
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• 2006

This study was undertaken to develop a reliable mice model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. BALB/C mice and NC/Nga mice were sensitized twice with $100{\mu}l$ of 1% 2,4-dinitrochlorobenzene (DNCB) or vehicle (acetone : olive oil=4:1 mixture) in a week and challenged twice with $100{\mu}l$ of 0.2% DNCB or the vehicle at the following week. Mice were sacrificed at 19 days following the second DNCB or vehicle challenge for NC/Nga mice and at 28 days following the second DNCB or vehicle challenge for BALB/c mice. Upregulation of plasma 1gE, a hallmark of atopic dermatitis occurrence, was evident in the plasma obtained 4 day after the second DNCB challenge from BALB/c mice (approximately 4-fold) and NC/Nga mice (approximately 6-fold) treated with DNCB in comparison with that of the vehicle treated-control mice, and remain higher $3{\sim}4$ week after the second challenge. Ratio of plasma IgG1 versus IgG2a concentration was significantly higher in the mice treated with DNCB than the control mice, which also implies the skewed type-2 reactivity in vivo. Ratio of interleukin-4 versus interferon gamma produced in the splenic T cell culture supernatants was approximately 3-fold higher in the both strains of mice treated with DNCB than their control mice, respectively. The DNCB-treated mice demonstrated atopic dermatitis-like skin legions characterized with erythma, scaling, and hemorrhage, which was not observed with the control mice. Scratching on face or dorsal area was significantly more frequent (approximately 25-fold) in the DNCB-treated mice than the control at next day of the second DNCB challenge, and scratching frequency remains higher (approximately 4-fold) in the mice treated with DNCB than the control at 14 day following the second DNCB challenge. Overall, the mice model developed through sensitization and challenge with DNCB may be useful for research on atopic dermatitis and development of treatment materials for atopic dermatitis.

• The Korea Journal of Herbology
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• v.24 no.4
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• pp.215-224
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• 2009

Objectives : Atopic dermatitis is a chronic inflammatory disease characterized by typically distributed eczematous skin lesion with pruritus, lichenification and dry skin. In this study, we performed to assess the therapeutic effects of co-treatment of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS, C&C) on the TNCB(trinitrochlorobenzene)-induced atopic dermatitis in NC/Nga mice, characterized by the onset of atopic dermatitis along with an increase the number of inflammatory cells and dysregulation of Th2 cytokines. Methods : Defined amount of CGB was sprayed on mice skin and CHS was simultaneously orally administrated to TNCB treated NC/Nga mice for 5 weeks. The immune cell types were caracterized by flow cytometry using each specific antibody. The amount of Th2 cytokines in serum and splenocytes culture supernatant were measured by ELISA. Results : Administration of C&C significantly reduced clinical dermatitis severity including pruritus, edema, eczematous and erythema. Histological findings indicated that the thickening of epidermis/dermis and dermal infiltration of inflammatory cells were dramatically reduced. Flow cytometry analysis showed that infiltrated immune cell numbers of CCR3+, B220+/IgE+, Gr-1+/CD11b+, and CD117+ were significantly reduced in C&C-treated dorsal skin lesion. Furthermore, T cell composition rate in PBMC was also dramatically decreased by the treatment. C&C greatly down-regulated production of Th2 cytokines including IL-4, IL-5, IL-13 in the serum. The down- regulatory effects of C&C on these Th2 cytokines production were also detected in CD3/ CD28 activated splenocytes. Conclusions : These results indicated that C&C is a plausible therapeutic agent for treatment of atopic dermatitis through regulating the Th2 skewed immune system.

• BMB Reports
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• v.45 no.9
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• pp.538-543
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• 2012

We evaluated the expression of the costimulatory molecules CD80 and CD83 and major histocompatibility (MHC) class II induced by 2,4-dinitrofluorobenzene (DNFB) in the RAW 264.7 macrophage cell line. In contrast to the previously reported effect of DNFB on dendritic cells, CD86 expression did not change. Furthermore, we observed that the CD83 expression level transiently increased and then decreased. Induction of CD80 and MHC class II molecule expression and a decrease in CD83 expression by DNFB in vitro were also confirmed in splenocytes of BALB/c and NC/Nga mice. However, DNFB did not influence CD83 expression in peritoneal $CD11b^+$ cells from BALB/c or NC/Nga mice. Detailed in vivo experiments and further studies on the possible contribution of $CD11b^+$ cells to induce atopic dermatitis (AD) would be helpful to attain a better understanding of AD pathogenesis.

• Biomolecules & Therapeutics
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• v.22 no.5
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• pp.438-444
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• 2014

Broussonetia kazinoki Siebold. (B. kazinoki) has long been used in the manufacture of paper in Asian countries. Although B. kazinoki leaves (BK) have been employed in dermatological therapy, use of BK has not been tested in patients with atopic dermatitis (AD). Using Nc/Nga mice, which are genetically predisposed to develop AD-like skin lesions, we confirmed the efficacy of BK in AD treatment. BK extract was applied topically to Dermatophagoides farinae-induced AD-like lesions in Nc/Nga mice, and the effects were assessed both clinically and by measuring skin thickness on the back and ears. We measured the effects of BK extract on plasma levels of IgE and IL-4. We also measured the ability of BK extract to inhibit the secretion of hTARC in HaCaT cells after stimulation by TNF-${\alpha}$ and IFN-${\gamma}$. We found that BK extract significantly reduced ear and dorsal skin thickness and the clinical signs of AD, as well as significantly down-regulating the plasma levels of IgE and IL-4 (p<0.01 for each comparison). Moreover, $500{\mu}g/mL$ of BK extract inhibited hTARC secretion in HaCaT cells by activated TNF-${\alpha}$/IFN-${\gamma}$ by about 87%. These findings suggest that topical application of BK extract has excellent potential in the treatment of AD.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.2
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• pp.116-122
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• 2019

The aim of this study is to evaluate the anti-inflammatory effect of Hataedock treatment using Coptidis Rhizome and Glycyrrhiza Uralensis (CG) mixed extract in allergic rhinitis induced NC/Nga mice. We divided NC/Nga mice into 3 groups as follows; allergic rhinitis-induced group after CG Hataedock treatment (CGT, n=10), no treatment group (Ctrl), allergic rhinitis elicited group (ARE). To induce allergic rhinitis, NC/Nga mice of 3 weeks age were sensitized on 7, 8 and 9week by Ovalbumin (OVA) antigen in intranasal space. Hataedock using CG extract was administered on week 3 in allergic rhinitis-induced group (CGT) after Hataedock treatment. To identify distribution of Interlukin (IL)-4, Cluster of differentiation 40 (CD40), high-affinity IgE receptor ($Fc{\varepsilon}RI$), substance P, Matrix metallopeptidase 9 (MMP-9), Nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) p65, Inducible nitric oxide synthase (iNOS) and Cycloxygenase-2 (COX-2), we used histological examination. CGT significantly inhibited IL-4 and CD40 response compared with ARE. The reduction of Th2 cytokine expression decreased inflammatory mediators such as $Fc{\varepsilon}RI$, substance P, MMP-9, $NF-{\kappa}B$ p65, iNOS and COX-2. Such immunological improvement induced reduction of respiratory epithelial damage and mucin secretion in goblet cell. These results indicate that Hataedock treatment suppresses allergic rhinitis through modulating of Th2 responses and diminishing various inflammatory mediators in nasal mucosal tissue. It might have potential applications for prevention and treatment of allergic rhinitis.