• Journal of Nutrition and Health
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• v.39 no.6
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• pp.520-528
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• 2006

Free amino acids in epidermis function as a major component of Natural Moisturizing Factor (NMF), which maintains the optimal level of water in skin even at the low humidity. In fact, the depletion of free amino acids is reported in the epidermis of atopic dermatitis, the skin condition involving dryness. As an effort searching the dietary source for improving the level of water and free amino acid in epidermis, the dietary effects of silk protein, sericin (S) and fibroin (F) on trans epidermal water loss (TEWL), and plasma and epidermal levels of free amino acids were compared in this study. Thirty of male NC/Nga mice, an animal model of atopic dermatitis, were divided into three groups: group CA as an atopic control with control diet, group S: 1% sericin diet and group F: 1% fibroin diet. Ten of male BALB/c mice were served as group C (control group) with control diet. All mice were fed on diet and water ad libitum for 10weeks. Dry skin condition was established in group CA as TEWL was increased (148.7% of group C). In parallel, epidermal level of glutamate, one of major amino acids functioning as NMF, was dramatically decreased and epidermal levels of methionine and alanine were inversely elevated. Dietary supplementation of sericin (group S) reduced TEWL at the similar level with group C and increased epidermal levels of glutamate as well as serine and glycine, the other major amino acids as NMF. Despite a marked decrease of methionine and alanine, the reduction of TEWL and epidermal levels of glutamate, serine and glycine of group F were less than of group S. Furthermore, in contrast to similar levels of other free amino acids in plasma and epidermis of group S and group C, plasma and epidermal levels of other free amino acids, specifically phenylalanine, isoleucine, cysteine and tyrosine in epidermis of group F, were significantly higher than of group C. Together, our data demonstrate that dietary supplementation of sericin is more effective at improving dry skin condition that paralleled with the normalization of free amino acids in plasma and epidermis of NC/Nga mice.

• The Journal of Pediatrics of Korean Medicine
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• v.30 no.4
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• pp.29-59
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• 2016

Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

• The Journal of Pediatrics of Korean Medicine
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• v.25 no.1
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• pp.1-27
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• 2011

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.

• IMMUNE NETWORK
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• v.8 no.3
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• pp.98-105
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• 2008

Background: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. Methods: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-${\gamma}$) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. Results: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-${\gamma}$ while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. Conclusion: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.

• Journal of Haehwa Medicine
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• v.17 no.2
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• pp.167-183
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• 2008

KCSYJT medicines controlled $CD4^+/IFN-\gamma$, and $CD4^+/CD25^+/foxp3^+$ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates T cells of a NC/Nga mouse same time by anti-CD40/rmIL-4, and interleukin-$1{\beta}$, IL-6, TNF-$\alpha$, and TGF-$\beta$ mRNA outturn that bear in T and B cells decreased remarkably by KCSYJT medicines. Intracellular staining of splenocytes anti-CD40/rmIL-4 plus rmIL-4 stimulated as described in a, assessed after 24 h, KCSYJT exerts a mainly immunosuppressive effect that acts at least partially through suppression of the transcription factor GATA3 expression in $CD4^+$ T cells. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result that Th1 cell and Th2 cell observe to be shifted by cytokine expression with GATA3 regulation by KCSYJT medicines could know that KCSYJT medicines can use usefully in allergy autoimmnune diease.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.866-874
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• 2006

Atopic dermitiis(AD) is a chronic inflammatory skin disease, which requires safe and effective medicinal therapy. Over production of Th2 cytokines and chemokines as well as IgE, which are mediated by highly activated immune cells, have been considered as pathologic factors in this disease. We found that Gamipaidok-san(GPDS), which is a traditional herbal medicine clinically prescribing for atopic dermitis patients in the hospital, has suppressive effects on the development of DNC8 induced dermatitis in Nc/Nga atopic model. Oral administration of GPDS at the concentration of 250 mg/Kg for 12 weeks significantly suppressed the clinical severity of the dermatitis including pruities, edema, eczematous and dryness. Histological examination revealed that thickness of dermis and epidermis were considerably reduced, and the number of infiltrated inflammatory immune cells including mast cells, CCR3+, and CD4+ T cells were decreased in the affected skin and ear, and consistantly, the number of CD3+/CCR3+ cells in Iymph nodes were decreased. The levels of Th2 cytokines produced by activated splenocyte from atopic mice were also down-regulated by GPDS. Furthermore, the serum levels of IgE were considerably reduced, which accompanied by a decrease in the number of B220+IgE+ B cells in the Iymph nodes. Taken together, these results suggested that oral administration of GPDS, a traditional herbal medicine, has suppressive effects on atopic dermitis of Nc/Nga mouse by the modulation of the immune system, therefore GPDS has potential as a natural therapeutic for treatment of atopic dermatitis.

• Toxicological Research
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• v.24 no.1
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• pp.17-22
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• 2008

Sopungsan (SS) is a traditional Korean decoction used for the treatment of dermatitis. The aim of this study is to confirm whether or not SS has a preventive effect on the development of atopic dermatitis in dinitrochlorobenzene-applied Nc/Nga mice. SS was administered orally to Nc/Nga mice, which led to the remarkable suppression of the development of dermatitis, as determined by a histological examination and the serum IgE levels. Moreover, SS inhibited the production of thymus- and activation-regulated chemokine (TARC) and its mRNA expression in a keratinocyte cell line, HaCaT, which had been stimulated with tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$). Activation of the nuclear factor-${\kappa}B$ (NF-${\kappa}B$) or activator protein-1 (AP-1) is one of key steps in the signaling pathways mediating induction of TARC. In this study, SS selectively suppressed NF-${\kappa}B$ activation which may be essential for TARC expression in $TNF-{\alpha}/IFN-{\gamma}$ treated keratinocytes. The inhibitory effect of SS on NF-${\kappa}B$ activation and TARC production might be associated with the anti-dermatitic effects of SS.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.3
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• pp.10-19
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• 2008

Objective : To investigate the effects of Radix Ophiopogon japonieus on atopic dermatitis, I prepared DNCB(2,4-dinitrochlorobenzen) induced atopic dermatitis NC/Nga mice and observed the mice by four ways; eye observation, the number of skin behavior times, histological changes of skin and cytokine(Total IgE, IL-4, $IFN-{\gamma}$). Methods : After prepare Radix Ophiopogon japonicus extract, DNCB induced atopic dermatitis NC/Nga mice were divided into three groups. The first is Control group which was intact group. The second is Medication group which was orally medicated Radix Ophiopogon japonicus extract one time a day for consecutive 5 days. The third group is Application group which was applied Radix Ophiopogon japonicus extract externally one time a day for consecutive 5 days. After that, the effect of Radix Ophiopogon japonicus on atopic dermatitis was observed. Statistical analysis was performed by using Kmskal-Wallis test and statistical significance was set at less than 5%. Results : 1. Radix Ophiopogon japonicus showed some in both Medication group and Application At observation of skin morphologic change, effects to prevent erythema reaction on skin group. 2. At the number of scratching behavior times, Radix Ophiopogon japonicus showed an effect to decrease scratching behavior times, but there was no statistical significance among three groups. 3. At skin tissue H-E stain, Radix Ophiopogon japonicus showed an effect to prevent skin epidermal tissue damages and also showed that it could keep the skin healthy in both Medication group and Application group. Especially in Application group, the skin of mouse showed almost normal recovery. 4. At cytokines, there was no statistical significance among three groups in IgE and IL-4. But Radix Ophiopogon japonicus showed an significant effect to suppress $IFN-{\gamma}$ in both Medication group and Application group. There was no significant difference between two groups. Conclusion : Radix Ophiopogon japonicus has some effects on atopic dermatitis in both internal medication and external application.

• Journal of Haehwa Medicine
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• v.16 no.1
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• pp.81-91
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• 2007

Objective : We wished to examine closely effect that Kami-KangHwalSan medicines used to atopy dermatitis disease patient get in atopy eruption control experimentally. Materials and Methods : Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. However, the exact etiology of AD has been unclear. To further explore the pathogenesis and treatment of AD, a suitable animal model is required. We found that skin lesions, which were chnically and histologically very simlar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Results and Conclusion : Kami-KangHwalSan medicines controlled CD3+/CD69+, CD4+/CD25+, B220+/IgE+, and B220+/CD23+ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates splenocytes of a NC/Nga mouse same t1me by PWM, and interleukin-4, eotaxin 2, CCR3, TARC mRNA outturn that bear in splenocytes decreased remarkably by Kami-KangHwalSan medicines. Th1 cell and Th2 cell observe to be shifted by secretion amount of IL-4 and IFN-$\gamma$ by Kami-KangHwalSan medicines could know that Kami-KangHwalSan medicines can use usefully in allergy autoimmnune diease.

• Journal of Haehwa Medicine
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• v.16 no.1
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• pp.93-105
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• 2007

Objective : We wished to examine closely effect that Kami-JeSeubUilYeongTang medicines used to atopy dermatitis disease patient get in atopy eruption control experimentally. Materials and Methods : Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. However, the exact etiology of AD has been unclear. To further explore the pathogenesis and treatment of AD, a suitable animal model is required. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result and Conclusion : Kami-jeseupwiryeongtang(JWRTK) medicines controlled CD3+/CD69+, CD4+/CD25+, B220+/IgE+, and B220+/CD23+ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates splenocytes of a NC/Nga mouse same time by PWM, and interleukin-4, eotaxin 2, CCR3, TARC mRNA outturn that bear in splenocytes decreased remarkably by Jeseupwiryeongtang-Kamibang(JWRTK) medicines. Th1 cell and Th2 cell observe to be shifted by secretion amount of IL-4 and IFN-$\gamma$ by Jeseupwiryeongtang-Kamibang(JWRTK) medicines could know that Jeseupwiryeongtang-Kamibang(JWRTK) medicines can use usefully in allergy autoimmnune diease.