• 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.30 no.3
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• pp.1-30
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• 2016

Objectives Previous studies have found out that Forsythiae Fructus (FF) extracts have anti-atopic activities by in vitro experiment. In order to understand more about FF extracts' benefit, we subdivided FF extracts depending on systematic fractionation method by using Methylene chloride (MC), Ethyl acetate (EtOAc), n-BuOH and n-hexane (n-Hx). This study is designed to examine the effect of FF fractions on the PMA- ionomycin-induced activation of RBL-2H3 mast cell lines in vitro and on the DNCB-induced activation of NC/Nga mice in vivo. Methods For this study, we examined IL-4, IL-13 production by ELISA analysis, IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$ mRNA expression by real-time PCR and manifestations of AP-1 and MAPKs transcription factors by western blotting in vitro. Through in vitro experiment, we selected FF n-BuOH fraction that seems the best effective in atopic dermatitis then induced it on NC/Nga mice by DNCB. We measured mice's WBC, eosinophil and neutrophil in heart blood, IL-4, IL-5, IFN-${\gamma}$ in the spleenocyte culture supernatant, the absolute cell numbers of CD4+, CD8+, B220+CD23+, CD3+CD69+ and Gr-1+CD11b+ in the PBMCs, ALN and dorsal skin, IL-5, IL-13, IL-31, IL-31RA in the dorsal skin by real-time PCR and the distribution of immune cells by H&E on dorsal skin and ANL and toluidine blue staining on dorsal skin. Results FF n-BuOH fraction suppressed IL-4, IL-13 production and mRNA expression of IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$. Results from the western blot analysis showed that FF n-BuOH fraction reduced the activation of the mast cell specific transduction factors involved in AP-1 by suppressing JNK and ERK phosphorylation. In the gross, atopic dermatitis induced by DNCB in NC/Nga mice were improved by oral administration of FF n-BuOH fraction. Oral FF n-BuOH fraction also decreased the level of IgE in mice's serum and the level of IL-4 and IL-5 in the spleenocyte culture supernatant, cell numbers of CD8+, B220+CD23+ in the PBMCs, CD4+ in the ALN and CD4+, Gr-1+CD11b+ in the dorsal skin and suppressed mRNA expression of IL-5, IL-13, IL-31, IL-31RA in the dorsal skin. Histological examination showed that infiltration levels of immune cells in atopic dermatitis induced NC/Nga mice were improved by FF n-BuOH fraction. Conclusions FF n-BuOH fraction can reduce pruritus by suppressing IL-31, IL-31RA secretion and modulate molecular mediators and immune cells associated with atopic dermatitis induced in NC/Nga mice which may have played a significant role in recovering atopic dermatitis symptoms.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.27 no.4
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• pp.141-157
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• 2014

Objectives : The purpose of this study is to investigate the effects of DOGO phreatic water containing germanium on Atopic Dermatitis in NC/Nga mouse. Methods : We made DOGO phreatic water added germanium. After making atopic dermatitis caused by sensitizing NC/Nga mouse to DNCB(dinitrochlorobenzene), we made mouse swim in tanks each filled with distilled water, tap water, DOGO phreatic water, DOGO phreatic water(added germanium) for 30minutes everyday. 3weeks later, we analyzed skin clinical score, total IgE levels(by ELISA), WBC differential counting(Neutrophils, Monocytes), absolute cell number of $Neutrophil^+Gr-1^+$, CCR3 mRNA expressions(by Real-time PCR), IL-4, IFN-${\gamma}$ production levels(by ELISA), histologic test(by H&E staining, toluidine blue staining). Results : The results of making NC/Nga mouse induced atopic dermatitis swim in tanks filled with DOGO phreatic water(contain germanium) are as follows. 1. Skin clinical scores were decreased significantly in comparison to control group. 2. Total IgG levels were decreased significantly in comparison to control group. 3. WBC differential counting(Neutrophils, Monocytes) were decreased significantly in comparison to control group. 4. Absolute cell number of $Neutrophil^+Gr-1^+$ were decreased significantly in comparison to control group. 5. CCR3 mRNA expressions were decreased significantly in comparison to control group. 6. IL-4, IFN-${\gamma}$ production levels were decreased significantly in comparison to control group. 7. The epithelial tissue thickness, leucocytes infiltration, erythema, edema, excoriation, scaling, mast cells infiltrations in dorsal skin were decreased in comparison to control group. Conclusions : These results indicate that DOGO phreatic water(contain germanium) can be used for helping treat atopic dermatitis.

• 한국약용작물학회:학술대회논문집
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• 2010.10a
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• pp.349-350
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• 2010

• The Journal of Pediatrics of Korean Medicine
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• v.35 no.4
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• pp.167-167
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• 2021

• 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 Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.181-189
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• 2007

To examine the effects of HHCP on atopic dermatitis and its various immunopathologic parameters was induced by DNCB in NC/Nga mice and the animals were orally administrated with HHCP. We summarized the results obtained from serum levels of IgE and the numbers of various immune cells as follow. HHCP has no cytotoxic effects at the range of concentration (1-400 ${\mu}g$/ml) on fibroblast isolated from lung of BALB/c mice. HHCP significantly lowered the serum levels of IgE compared with control at 16 and 20 week. HHCP significantly reduced the number of CD19$^+$ cell in spleen and DLN, as well as the number of B220$^+$ /IgE$^+$ cell in DLN compared with control. HHCP significantly reduced the number of ${\alpha}$${\beta}$ TCR$^+$ in spleen and DLN, the number of CD8$^+$ in spleen compared with control, and also significantly reduced the number of CD3$^+$, CCR3, CD3$^+$/CD69$^+$, CD3/ CCR3, CD4$^+$, CD3$^+$/ CD4$^+$/CD45$^+$ cell in DLN. HHCP increased the number of NK$^+$ cells in spleen compared with control, in contrast significantly decreased the number of CD11c$^+$/ Classll$^+$ cell and CD11b$^+$/Gr-1$^+$ cell in DLN. Taken together, these results suggested that HHCP has suppressive effects on atopic dermatitis through the inhibition of IgE production and modulation of immune cell population in NC/Nga mice.

• 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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.5
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• pp.1099-1107
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• 2007

This study was investigated the anti-allergic effect of SJSBT on DNCB induced atopic dermatitis in NC/Nga mice. We summerized as the follow. SJSBT significantly decreased the clinical manifestations of atopic dermatitis including itching, dryness, edema, hemorrhage, and lichenification in dose dependent manner. SJSBT markedly suppressed invasion and edema of leukocytes and mast cell in dorsal skin and ear tissue. SJSBT significantly reduced the number of CD11b+/Gr-1 cell compared with positive control, but it had no affect the number of CD3+ and CCR3+/CD3+ cell. SJSBT markedly suppressed the expression of cytokine and chemokine such as IL-6, $TNF-{\alpha}$, eotaxin and CCR3 compared with positive control group. SJSBT significantly decreased the invasion of CD4+ and CCR3+ cell in ear and dorsal skin tissue compared with positive control group by using immunohistochemical staining. Taken together, these findings suggested that SJSBT has an anti-allergic activity and this might be useful for the clinical application to treat allergic diseases such as atopic dermatitis.