• 혜화의학회지
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• 제16권2호
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• pp.251-265
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• 2007

Yanhyeoljeseuptang(YHJST) is a traditional herbal medicine used for the treatment of dermatitis. The aim of this study was to confirm whether or not YHJST has a preventive effect on development of atopic dermatitis in dinitrochlorobenzene(DNCB)-applied Nc/Nga mouse. This study was undertaken to develop a reliable mouse model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. NC/Nga mouse were sensitized with $200\;{\mu\ell}$ of 1% 2,4-dinitrochlorobenzene(DNCB) (acetone : olive oil = 3 : 1 mixture) and challenged twice or three times with $150\;{\mu\ell}$ of 0.2% DNCB in a week for the following 4 weeks. YHJST was administered orally to Nc/Nga mouse for 8 weeks, which led to the remarkable suppression on the development of dermatitis, as determined by various immune factors related to pathogenesis of atopic dermatitis in splenocytes and DLN cells. In this study, YHJST selectively suppressed T ce11 (CD4+, CD3+/CD69+, CD4+/CD25+) activation, which may be essential for ratio of IL-4 versus INF-$\gamma$ produced in the splenic T cell culture supernatants was approximately 3-fold higher in the mouse treated with DNCB than their control mouse respectively. Immunologic studies showed down-regulated that the capacity of spleen T cells to produce IL-4, but IFN-$\gamma$ was up-regulated by means of oral intake of these YHJST. These results strongly suggest that YHJST is a promising candidate for treatment of human atopic dermatitis.

• 혜화의학회지
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• 제16권2호
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• pp.267-280
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• 2007

Gupoongjeseuptang(GPJST) is a traditional herbal medicine used for the treatment of dermatitis. The aim of this study was to confirm whether or not GPJST has a preventive effect on development of atopic dermatitis in dinitrochlorobenzene(DNCB)-applied Nc/Nga mouse. This study was undertaken to develop a reliable mouse model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. NC/Nga mouse were sensitized with $200\;{\mu\ell}$ of 1% 2,4-dinitrochlorobenzene(DNCB) (acetone : olive oil = 3 : 1 mixture) and challenged twice or three times with $150\;{\mu\ell}$ of 0.2% DNCB in a week for the following 4 weeks. GPJST was administered orally to Nc/Nga mouse for 6 weeks, which led to the remarkable suppression on the development of dermatitis, as determined by various immune factors related to pathogenesis of atopic dermatitis in splenocytes and DLN cells. In this study, GPJST selectively suppressed T ce11 (CD4+, CD3+CD69+, CD4+CD25+) activation, which may be essential for ratio of IL-4 versus INF-$\gamma$ produced in the splenic T cell culture supernatants was approximately 3-fold higher in the mouse treated with DNCB than their control mouse respectively. Immunologic studies showed down-regulated that the capacity of spleen T cells to produce IL-4, but IFN-$\gamma$ was up-regulated by means of oral intake of these GPJST. These results strongly suggest that GPJST is a promising candidate for treatment of human atopic dermatitis.

• 대한한방소아과학회지
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• 제23권2호
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• pp.51-85
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• 2009

Objectives : The purpose of this study is to investigate the effect of concurrent administration of KKSDU and AJ on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the condition in humans. Methods : We evaluated clinical skin score, hematology, serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse and analyzed the cytoline level, total cell number, immunohistochemical staining, histological features of axillary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results : Orally administration of KKSDU and concurrent administration of KKSDU and AJ decreased the clinical skin score, total cell number of WBC, platelet, neutrophils, eosinophils in blood, serum total IgE & IgG1, IL-5, IL-13. Also, total cell number of ALN and dorsal skin tissue, absolute cell number of CD3e+&CD19+, CD4+&CD8+, CD3+/CCR3+, CCR3+, CD3+/CD69+, CD3+/CXCR5+ in ALN, PBMCs, absolute cell number of CCR3+, CD3+/CD69+, CD11b+/Gr-1+ in dorsal skin tissue, Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue and gene expression of IL-5 mRNA, IL-13 mRNA in ALN are significantly decreased. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell & mast cell in dermis, histologic infiltration of mast cell, the size of inflammatory lymphocytes cells & plasma cells in ALN and histologic infiltration of CD4+ & CCR3+ in ALN and dorsal skin tissue are significantly decreased. However, total cell number of DLN, absolute cell number of CD3+&CD19+, CD4+&CD8+, B220+/CD23+, CD3+/CD69+ in DLN and CD4+CD25+foxp3+Treg cell, foxp3 mRNA in dersal skin tissue are increased significantly. Conclusions : Concurrent administration of KKSDU and AJ on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse was very effective to the atopic detmatitis treatment.

• 대한한방소아과학회지
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• 제22권3호
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• pp.105-137
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• 2008

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.

• 대한한방소아과학회지
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• 제23권1호
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• pp.37-72
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• 2009

Objectives The purpose of this study is to investigate the suppressive effects of Atopy cream-combined with Jawoongo ointment (A-J), on the development of atopic dermatitis-like skinlesions in NC/Nga mouse. Methods We evaluated clinical skin score, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, analyzed the cytokine level, total cell number, Immunohistochemical staining and Histological features of axillary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results A-J decreased the clinical skin score, total cell number of WBC, platelet, neutrophils, eosinophils in blood, Serum total IgE & IgG1, IL-5, IL-13. Also, total cell number of ALN and dorsal skin tissue, Absolute cell number of $CD3e^+$&$CD19^+$, $CD4^+$&$CD8^+$, $CD3^+/CCR3^+$, $CCR3^+$, $CD3^+/CD69^+$, $CD3^+/CXCR5^+$ in ALN, PBMCs, Absolute cell number of $CCR3^+$, $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ in dorsalskin tissue, Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue and gene expression of IL-5 mRNA, IL-13 mRNA in ALN decreased significantly. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell and mast cell in dermis, histologic infiltration of mast cell, the size of inflammatory lymphocytes cells and plasma cells in ALN and histologic infiltration of CD4+ & CCR3+ in ALN and dorsal skin tissue decreased significantly. However, total cell number of DLN, absolute cell number of $CD3e^+$&$CD19^+$, $CD4^+$&$CD8^+$, $B220^+/CD23^+$, $CD3^+/CD69^+$ increased significantly. Conclusions A-J was the successful treatment of atopic dermatitis in a NC/Nga mouse model.

• 대한한방소아과학회지
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• 제22권2호
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• pp.81-114
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• 2008

Objectives : The purpose of this study is to investigate the effect of Jeseupwiryeongtang-gagam(JWTG) 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 JWTG on AD, 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 and Conclusions : In vivo, clinical skin severity score were significantly lower in JWTG group than control group. IgE, IL-6, $TNF-{\alpha}$, IgM, IgG2a and IgG2b levels in serum were decreased remarkably in JWTG group than control group and $IFN-{\gamma}$ production, secreted in Th1 cell were increased by JWTG. After experiment ended, we analyzed immunological cells ($CD3^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3^+$$CD69^+$, $CD4^+$$CD25^+$ and $CD49b^+$) by flow cytometry. It resulted that total absolute number of $CD3^+$, $CD19^+$, $CD4^+$ and $CD8^+$ cells were recovered as normal and $CD3^+$$CD69^+$ were decreased significantly compared with control group in isolated DLN and PBMCs from NC/Nga mouse and total absolute number of $Gr-1^+$, $CD11b^+$ and $CD3^+$ in dorsal skin of NC/Nga mouse were decreased by JWTG. We analyzed ear, DLN, and neck-back skin after biopsy and dyeing by hematoxyline/eosin(H&E) and toluidine staining (mast cells marker) and obtained results that JWTG were effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell infiltration). Ear thickness was decreased significantly than the control group and the size of inflammatory lymphocytes cells(ILC) and plasma cells(PC) in DLN were also decreased.

• 대한한방소아과학회지
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• 제22권1호
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• pp.69-93
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• 2008

Objectives The purpose of this study is to investigate the effect of KKHS on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to this condition in humans. Methods To investigate the effect of KKHS on atopic dermatitis (AD), 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 of NC/Nga mouse in vivo. Results In vivo, clinical skin severity score was significantly lower in the KKHS group than in the control group. IgE, IL-6, TNF-${\alpha}$, IgM, IgG2a and IgG2b levels in serum decreased remarkably in the KKHS group than in the control group, and the level of IFN-${\gamma}$ production which is secreted from Th1 cell was increased by KKHS. After this experiment we analyzed immunological cells ($CD3^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3^+CD69^+$, $CD4^+CD25^+$ and $CD49b^+$) by flow cytometry. It results that the total absolute number of $CD3^+$, $CD19^+$, $CD4^+$ and $CD8^+$ cells were recovered as much as normal state, and the level of $CD3^+CD69^+$ in isolated DLN and PBMCs were significantly decreased, and total absolute number of $Gr-1^+$, $CD11b^+$ and $CD3^+$ in dorsal skin of NC/Nga mouse were decreased by KKHS. We analyzed ear, DLN, and neck-back skin after biopsy and dyeing by hematoxyline/eosin(H&E), toluidine staining (mast cells marker). KKHS were very effective to the histological symptoms which are in dermal and epidermal thickening, hyperkeratosis and inflammatory cell infiltration. Ear thickness was significantly decreased compared with the control group and the size of inflammatory lymphocytes cells (ILC) and plasma cells (PC) in DLN were also decreased. Conclusions KKHS on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse was very effectiveness to the atopy dermatitis treatment.

• 대한한방소아과학회지
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• 제25권1호
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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.