• Journal of Acupuncture Research
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• v.37 no.2
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• pp.123-127
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• 2020

Background: This study was designed using a mouse model of atopic dermatitis [phthalic anhydride (PA)-treated mice], to investigate the anti-inflammatory effect of bee venom pharmacopuncture (BVP) in keratinocytes. Methods: Western blot analysis was performed to investigate inflammation related protein expression of iNOS, COX-2, phospho-ERK (p-ERK), and ERK, in LPS (1 ㎍/mL)-activated keratinocytes, following BVP treatment, and in PA-treated mice, after BVP treatment. Griess reaction was performed to investigate NO concentration. Enzyme-linked immunosorbent assays were used to determine the concentrations of interleukin (IL)-4+, IL-17A+, IL-13 and IL-4 in PA-treated mice after BVP treatment. In addition, monocyte, macrophage, neutrophil, and eosinophil counts were measured to observe the changes in white blood cell infiltration. Results: The keratinocytes of the BVP-treated group showed a decreased expression of iNOS, COX-2, ERK at 5 OX-2, ERK E, and p-ERK at 1, 2 and 5 RKRK ERK ERK, and a dose-dependent decrease in NO concentration at 2 and 5 ntrationof s. In the BVP-treated groups (0.1 μ.1-trea μ.1-treated gr), PA-treated mice showed recovery after 4 weeks which was dose-dependent, showing a significant decrease in clinical scores for AD, and a decreased concentration of IL-13 and IL-4 with BV treatment. There was a dose-dependent decrease in the infiltration of eosinophils, neutrophils, monocytes, macrophages, and a decreased thickness of the epidermis due to inflammation, and decreased expressions of iNOS, COX-2, p-ERK, ERK, especially in the 0.1 μ0/mL BVP-treated group, Conclusion: These results suggest that BVP may be an effective alternative treatment for atopic dermatitis.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.32 no.2
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• pp.94-106
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• 2019

Objectives : This study was conducted to investigate the effects of Gwakhyangjeonggi-san(GJS) on atopic dermatitis(AD) induced by 2,4-dinitrochlorobenzene(DNCB) in mice. Methods : The mice(Balb/c mice) were divided into three groups; normal Balb/c mice with oil treatment(Sham group), DNCB-induced AD mice(AD group), and GJS treated AD mice(GJS group). GJS group were orally administered GJS daily for 2 weeks. We observed changes of clinical skin severity score, the expression of thymic stromal lymphopoietin(TSLP), interleukin(IL)-4 and tumor necrosis factor(TNF)-${\alpha}$ in skin and mast cell infiltration. Also, serum immunoglobulinE(IgE), IL-4, $TNF-{\alpha}$ and IL-6 were evaluated. Results : The clinical skin severity score of GJS group was decreased compared to AD group. In hematoxylin and eosin staining results, GJS group showed a significant reduction of epithelial skin thickness. In addition, expression of TSLP and mast cell infiltration in skin were also reduced by GJS treatment compared to those of AD group. Thus, we evaluated expression of IL-4, Th2-dependent cytokine, and $TNF-{\alpha}$, pro-inflammatory cytokine in skin. GJS significantly reduced both IL-4 and $TNF-{\alpha}$ compared to AD mice. Moreover, levels of IgE, IL-4, $TNF-{\alpha}$ and IL-6 in plasma also significantly decreased by oral GJS treatment. Conclusion : The present study suggests that GJS can significantly reduced symptoms of AD, therefore it can be a promising candidate for anti-atopic dermatitis treatment.

• The Journal of Pediatrics of Korean Medicine
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• v.36 no.3
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• pp.87-96
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• 2022

Objective The purpose of this study was to confirm the effect of Sabaek-san extract on skin damage recovery and inflammation relief in atopic dermatitis-induced mice through Endocannabinoid system (ECS) control. Methods In this study, we used 6-week-old NC/Nga mice were divided into 4 group: control group (Ctrl), lipid barrier elimination group (LBEG), palmitoylethanolamide (PEA) treated group after lipid barrier elimination (PEAG), and Sabaek-san extract treatment group after lipid barrier elimination (SBSG). Each group was assigned 10 animals. After drug administration of three weeks duration following lipid barrier elimination, cannabinoid receptor (CB) 1, CB2, CD (Cluster of Differentiation) 68, phosphorylated inhibitor kappa B (p-IκB), inducible nitric oxide synthase (iNOS), Fc ε receptor, substance P and serotonin were observed to confirm the regulation of the ECS, macrophage activity and mast cell activity. Results We found that 8-hydroxydeoxyguanosine (8-OXdG) positive reaction was significantly lower in the SBST group than in LBET and PEAT groups. Both CB1 and CB2 showed higher positive reactions in the SBST group than in the LBET and PEAT. CD68, p-IκB, iNOS, Fc ε receptor, Substance P and serotonin showed lower positive reaction in the SBST compared to the LBET and PEAT. Conclusion It was confirmed that the Sabaek-san extract can reduce the inflammation of atopic dermatitis by restoring the structural damage of the skin lipid barrier through ECS activity.

• 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.

• Food Science and Preservation
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• v.19 no.5
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• pp.792-798
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• 2012

The purpose of this study was to evaluate the anti-atopic-dermatitis effect of Angelica keiskei extract using a DNFB-induced animal model of atopic-dermatitis symptoms. A. keiskei was prepared via extraction with DW, 50% ethanol, and 100% ethanol in addition to fresh juice. All the extracts reduced the thickness levels of the ears and ear epidermis against swelling by DNFB inducement, especially in the aqueous-extract-treated group. The mRNA expressions of matrix metalloproteinases (MMPs), however, were not observed, which may indicate that Angelica keiskei extract alleviates the atopic-dermatitis symptoms in an MMP-independent manner. Finally, the level of the inflammatory cytokine IL-4 was inhibited only in the juice-treated group, although the IL-13 level was inhibited in the juice-, 50%-ethanol-extract-, and 100%-ethanol-extract-treated groups in a dose-dependent manner, and was not inhibited in the aqueous-extract-treated group. Taken together, these results suggest that Angelica keiskei extract has an inhibitory effect on atopic dermatitis, and may be a useful biomaterial for the development of cosmeceuticals.

• Journal of Physiology & Pathology in Korean Medicine
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• v.36 no.5
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• pp.187-192
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• 2022

The purpose of this study is to examine the effect of herbal medicine complex (HMC) containing Camellia sinensis L., Duchoesna chrysantha, Houttuynia cordata Thunberg, Poncirus trifoliata Rafinesque on skin inflammation and atopic dermatitis. First, we examined the anti-inflammatory effect of HMC in TNF-α induced human keratinocytes (HaCaT cell). Real-time PCR and western blotting were performed to evaluate the expression of inflammatory cytokines (e.g., iNOS, COX-2, IL-6, IL-8) mRNA and protein. Four-weeks old male NC/Nga mice were treated with 1% 2,4-dinitrochlorobenzene (DNCB) solution and used as an atopic dermatitis mice model. And, HMC (200 mg/kg or 400 mg/kg) was administered directly into the stomach of mice for 4 weeks, and blood or serum analysis, tissue staining were performed after oral gavage. As a result HMC inhibited the mRNA expression of iNOS, COX-2, IL-6, and IL-8, which had been increased by TNF-α in HaCaT cells. In addition, the protein expression was also significantly suppressed in the same way as the mRNA expression results. The in vivo experiment results showed that, HMC administration reduced thickening of the epidermis and infiltration of eosinophil into the skin stratum basale compared to DNCB treatment. In addition, HMC administration significantly reduced the inflammatory cytokines (IL-4, IL-5, IL-6, and IL-13) production and immunocyte (white blood cell, lymphocyte, neutrophil, and eosinophil) count compared to DNCB treatment. Moreover, the serum IgE and histamine level was decreased by HMC administration. These results suggest that HMC can be used as effective herbal medicine extract for skin inflammation and atopic dermatitis. And this study may contribute to the development of the herbal medicine-based drug for the treatment of skin inflammation and atopic dermatitis.

• Natural Product Sciences
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• v.21 no.3
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• pp.205-209
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• 2015

Fucoidan, a sulfated polysaccharide is found in several types of edible brown algae. It has shown numerous biological activities; however, the molecular mechanisms on the activity against atopic dermatitis have not been reported yet. We now examined the effects of fucoidan on chemokine production co-induced by TNF-α/IFN-γ, and the possible mechanisms underlying these biological effects. Our data showed that fucoidan inhibited the TNF-α/IFN-γ-induced production of thymus and activation-regulated chemokine (TARC) and macrophagederived chemokine (MDC) mRNA in human keratinocytes HaCaT cells. Also, fucoidan suppressed phosphorylation of nuclear factor kappa B (NF-κB) and activation of signal transducer and activator of transcription (STAT)1 in a dose-dependent manner. In addition, fucoidan significantly inhibited activation of extracellular-signal-regulated kinases (ERK) phosphorylation. These data indicate that fucoidan shows anti-inflammatory effects by suppressing the expression of TNF-α/IFN-γ-induced chemokines by blocking NF-κB, STAT1, and ERK1/2 activation, suggestive of as used as a therapeutic application in inflammatory skin diseases, such as atopic dermatitis.

• BMB Reports
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• v.55 no.6
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• pp.275-280
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• 2022

The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.

• Journal of Physiology & Pathology in Korean Medicine
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• v.36 no.1
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• pp.23-27
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• 2022

This study aims to evaluate the anti-inflammatory effect of 80% ethanol extracts of Stachys affinis (MQ) on 1-fluoro-2,4-Dinitrofluorobenzene (DNFB) induced atopic dermatitis (AD) in Balb/c mice. AD-like allergic contact dermatitis was induced by challenge of DNFB on the ear after DNFB sensitization on the back sides of mice. MQ alleivated clinical severity in AD-like skin lesions. In addition, ear thickness of epidermis and penetration of inflammatory cells in AD-like skin lesions were decreased by topical application of MQ. The the serum levels of tumor necrosis factor-α (TNF-α) and interleukin-4 (IL-4) were measured in AD mice using ELISA kits. Levels of TNF-α and IL-4 in serum were significantly decreased by topical application of MQ. Therefore, this study could give a clinical basis that MQ could be a agent to prevent AD.

• Journal of Physiology & Pathology in Korean Medicine
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• v.34 no.6
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• pp.341-347
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• 2020

The purpose of this study was to investigate the possibility of herbal medicine complex extract (NI-01), which were prepared from 6 natural materials (Cinnamomum cassia Blume, Lonicerae Flos, Paeonia suffruticosa Andrews, Arctium lappa Linne, Schzandra chinesis Bailon, Elsholtzia ciliata Hylander), as a functional material for inhibition of atopic dermatitis. anti-oxidative activity was confirmed by measuring DPPH electron donating ability and ABTS+ radical scavenging ability. Cytotoxicity and NO inhibition were measured using RAW 264.7 cells to confirm anti-inflammatory efficacy. The test substance was orally administered to the pruritus-induced ICR mice to confirm the inhibition of pruritus. The bovine cornea opacity and permeability (BCOP) assay was performed to confirm safety for irritation. NI-01 showed high antioxidant activity in DPPH and ABTS+ methods. In the anti-inflammatory effect tests with RAW 264.7 cells, NO production was inhibited at NI-01 concentrations of 50 (14.9%) and 100 (4.2%) ㎍/mL, which indicated that the anti-inflammatory effect was increased in a concentration-dependent manner. NI-01 also showed anti-itching effect after inducing of itching by compound 48/80 in ICR mice. NI-01 was proved to be a non-irritant substance in BCOP assay. The results of this study suggested that the herbal medicine combined extract (NI-01) has high antioxidant, anti-inflammatory and anti-itching effects, and safety for irritation. Therefore, herbal medicine complex extract (NI-01) is thought to be highly applicable for the inhibitory ingredients of the atopic dermatitis.