• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.9
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• pp.1263-1269
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• 2010

This study was performed to evaluate the antioxidant activities and anti-asthma effects of Morus bark water extracts. Inhibitory effect of Morus bark onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activities in vitro. Anti-asthma activities of Morus bark water extracts were assessed by testing their effects on the degranulation of mast cell. For this, $\beta$-hexosaminidase released from a basophilic cell line, RBL-2H3 was used and pro-inflammatory cytokines were measured by ELISA kit. The antioxidant activities of water extracts of Morus bark was 59.2% in the DPPH assay at $2,000\;{\mu}g/mL$ and 78.8% in the hydroxyl radical scavenging assay at $2,000\;{\mu}g/mL$. Our results indicated that Morus bark water extracts effectively inhibited free radical generation. Morus bark water extracts inhibited inflammation-mediating substances such as histamine and $\beta$-hexosaminidase release from RBL-2H3 cells. Cytokine release demonstrated a more effective blockading ability of the Morus bark water extracts to the release of IL-4 and TNF-$\alpha$ compared to control. These results demonstrate that Morus bark may be beneficial in the treatment of allergic inflammatory disease.

• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.331-337
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• 2009

Interleukin-13 (IL-13) has been proposed as a therapeutic target for bronchial asthma as it plays crucial roles in the pathogenesis of the disease. We developed an in vitro test system measuring transcriptional downregulatory activities on IL-13 as a primary screening method to select drug candidates from natural products. The promoter region of IL-13 (-2,048 to +1) was cloned into the upstream of a luciferase gene in the plasmid pGL4.14 containing the hygromycin resistance gene as a selection marker, generating pGL4.14-IL-13. The EL-4 thymoma and RBL-2H3 mast cells transiently expressing this plasmid highly produced the luciferase activities by responding to PI (PMA and ionomycin) stimulation up to 8-fold and 13-fold compared with the control, respectively, whereas cyclosporin A, a well-known antiasthmatic agent, significantly downregulated the activities. The BF1 clone of RBL-2H3 cells constitutively expressing pGL4.14-IL-13 was established by selecting surviving cells under a constant lethal dose of hygromycin treatment. The feasibility of this system was evaluated by measuring the downregulatory activities of 354 natural products on the IL-13 promoter using the BF1 clone. An extract from Morus bombycis (named TBRC 156) significantly inhibited PI-induced luciferase activities and IL-13 mRNA expression, but not the protein expression. Fisetin (named TBRC 353) inhibited not only PI-induced luciferase activities and mRNA expression, but also the IL-13 protein secretion, whereas myricetin (named TBRC 354) could not suppress the IL-13 expression at all. Our data indicated that this in vitro test system is able to discriminate the effects on IL-13 expression, and furthermore, that it might be suitable as a simple and time-saving primary screening system to select antiasthmatic agents by measuring transcriptional activities of the IL-13 promoter.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.23 no.2
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• pp.118-151
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• 1997

Allergic contact dermatitis is a common skin disease resulting from specific immumologic sensitization to topically applied various allergen. Purpose of this study was to investigate skin morphologic chganges by light microscopic, changes of protein band by SDS-PAGE in the skin treated on the surfactant. Furthermore, lectin histochemistry is used to know the chagnges of the terminal sugar of the glycoconjugate in the skin treated on the surfactant. The results were as follows : The 1 day of treated group was indicated the enlargement of capillary, the 3 days of treated group was showed that the cytoplasm was eosinophlic by the pyknotic of nucleus. The 6 days of treated group was observed hyperkeratinoid, increased of inflammation cells to epithelium. The 9 days of treated group was appeares that serum crust was sheded and formed a acanthosis. But dermis layer was cytoplasmic vaculation and enlargement of intercellular space. The 12 days treated group made up new epidermis layer of seven layer and of observed an decreased in quantity of inflammation cells. The mast cell of degranulated type was increased in treated surfactnat. It was secreating granules hold histamin, serotonin and heparin. Accordingly, protein band of electrophortic phase was observed a sudden changes since 3 days treated group. At observeation of the cell-surface glycoconjugates, LCA, PNA, SBA and WGA lectin positive cells but inducated lectin negative cells in Con-A. Furthermore, PNA and SBA showed stroger positive reaction as treated surfactant group.

• Journal of Haehwa Medicine
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• v.22 no.1
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• pp.145-170
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• 2013

Objectives : This study was carried out to investigate the anti inflammatory and anti allergy effects of Vitex rotundifolia Linne fil. extract(VRE). Results : 1. In vitro test, VRE was used to determine the modulation of cytokine secretion, the activation of inflammatory and allergic factor and the inhibition of gene expression. The cell survival rate of Raw 264.7 and Jurkat T cells didn't decrease and accordingly cytotoxicity wasn't observed. In anti-allergic assay, the secretion of IL-2, TNF-${\alpha}$, IL-4, IL-5 and IFN-${\gamma}$ were suppressed on Jurkat T cells induced by dust mites. And the gene expression of COX-2 was suppressed in HMC-1 stimulated by calcium ionophore A23187. In anti-inflammatory assay, the gene expression of TNF-${\alpha}$, COX-2 were suppressed on LPS-activated Raw 264.7 cells. And the secretion of IL-6 and IL-8 were suppressed on EoL-1 cells induced by dust mites. P38 and ERK activation of MAPK decreased generally. VRE showed potent inhibitory activity of NO production. 2. In vivo test, we used NC/Nga mouse induced by atopic dermatitis to observe the effects of VRE on the weight, water and feed, blood test, weight of organs, total IgE and histological change of main organs. Quantity of water and feed were not changed, therefore it didn't affect the weight directly, and no change was observed in related main organs, thus maybe there is no organ toxicity by test substances. And the symptoms were decreased significantly, and the thickness of epithelial cell layer and the number of mast cells were inhibited significantly by the difference of dosage. The number of total complete blood cells and IgE in serum were not changed significantly. Conclusion : These results suggest that VRE has anti-inflammatory and anti-allergic effects. Therefore VRE could be used effectively on improvement or treatment of atopic dermatitis. However, further study is needed to prove which component of VRE indicates effective pharmacological action.

• The Journal of Pediatrics of Korean Medicine
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• v.22 no.2
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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.

• Journal of Veterinary Clinics
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• v.26 no.5
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• pp.433-440
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• 2009

High levels of inflammatory cytokines were proposed contributors to the pathogenesis of a various inflammatory skin disorders. Therefore, investigating the immune response of the inflammatory skin disorder allows a better understanding of pathogenesis of a various inflammatory skin disorders and therapeutic approaches. The aim of this study was to analyze of the immune response in dogs with chronic inflammatory skin disease. To this aim, the present study evaluated relative mRNA expression of canine $IFN-{\gamma}$, IL-4, $TGF-{\beta}$ and IL-10 using TaqMan realtime PCR assays and semi-quantitative RT-PCR in freshly isolated peripheral blood mononuclear cells from twenty dogs with chronic inflammatory skin disease and ten normal dogs. The relative mRNA expression levels of IL-4 mRNA were significantly higher in dogs with chronic inflammatory skin disease than those in normal dogs (P < 0.01). The results of present study also showed a tendency towards increased expression of IL-10 transcripts in dogs with chronic inflammatory skin disease. However, there were no significant differences in the levels $IFN-{\gamma},\;TGF-{\beta}$ between normal and chronically inflammed dogs. In addition, the concentration of serum IgE was significantly increased in dogs with chronic inflammatory skin disease compared with those in normal dogs (P < 0.01). In histopathological examination, we found that there were markedly increased mast cell counts in chronically inflammed dogs (P < 0.05). These results suggest that the pathogenesis of chronic inflammatory skin disease might be associated with a T-cell mediated inflammatory responses characterized by a Th2-skewed immune response. Based on these results, the modulation of Th1/Th2 balance may be an effective therapeutic strategy for the treatment of chronic inflammatory skin disease.

• Journal of Life Science
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• v.19 no.12
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• pp.1808-1814
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• 2009

In our previous study, a new herbal preparation (HemoHIM) was developed as a functional food for the radioprotection and immunomodulatory agents. In order elucidate the mechanism involved, we examined the effect of HemoHIM on the compound 48/80-induced histamine release, and on the phorbol 12-myristate 13-acetate (PMA)/calcium ionophore (A23187)-induced inflammatory cytokine secretion in HMC-1. The cell culture supernatants were harvested, and the cytokines (IL-4, IL-6, IL-8, TNF-$\alpha$, GM-CSF) in the supernatants were measured by enzyme-linked immunosorbent assay. The total RNA of the cells was extracted, and the cytokines or c-kit/tryptase/Fc$\varepsilon$RI's messenger RNA expressions were examined using reverse transcriptase polymerase chain reaction. Under low concentrations, HemoHIM inhibited histamine release in HMC-1 stimulated compound 48/80. Furthermore HemoHIM inhibited PMA/A23187-induced inflammatory cytokines' secreation or mRNA expression in a dose-dependent manner. But IL-8 secretion was not inhibited by low concentrayion of HemoHIM, respectively. The mRNA expression of c-kit and Fc$\varepsilon$RI were also inhibited in a dose-dependent manner. Tryptase mRNA expression was only inhibited by low concentration of HemoHIM. These results indicated that HemoHIM might be an useful agent for protection against allergy as well as immune modulation, especially since it is a relatively nontoxic natural product.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.3
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• pp.51-66
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• 2005

Purpose : Investigate the effects of Haeganjeon water extract (HGJ) on immobilization-stress or cold-stress in C57BL/6J mice. Methods : Male C57BL/6J 30 mice of weighting 18${\pm}$2g, were divided into sixs groups including the immobilization-stress group(5heads), after immobilization-stress HGJ oral administration(500mg/kg) groups(5heads), cold-stress group(5heads) and after cold-stress HGJ oral administration(500mg/kg) groups(5heads). then we observed changes in the serum histamine and corticosterone level and changes immune system. Results : HGJ decreased the serum level of histamine and corticosterone increased by immobilization-stress or cold-stress. HGJ inhibited the release of histamine from mast cells. In addition, HGJ enhanced the cell viability of thymocytes decreased by immobilization-stress or cold-stress and decreased DNA fragmentation of thymocytes increased by immobilization- stress or cold-stress. Also, HGJ increased the cell viability of splenocytes decreased by cold-stress and decreased DNA fragmentation of splenocytes increased by cold-stress. HGJ decreased the population of thymic CD4+ cells increased by immobolization-stress. HGJ increased the population of B220+ cells decreased by immobilization-stress and decreased the population of Thy1+ cells increased by immobilization-stress. Also, HGJ decreased the population of splenic CD4+ cells increased by immobolization-stress. HGJ enhanced the production of ${\gamma}-interferon$ decreased by immobilization-stress or cold-stress and increased the production of interleukin-4 decreased by immobilization-stress. Furthermore, HGJ enhanced the phagocytic activity decreased by immobilization-stress or cold-stress and enhanced the level of nitric oxide decreased by immobilization-stress or cold-stress. Conclusion : HGJ may be useful for the prevention and treatment of stress via suppression of serum histamine and corticosterone level and enhancement of immune response.

• Journal of Life Science
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• v.17 no.11
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• pp.1511-1516
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• 2007

Basophils and mast cells play an important role in $Fc{\varepsilon}RI-mediated$ allergic reaction as effector cells. We studied the effects of Acanthopanax chiisanensis on $Fc{\varepsilon}RI\;{\alpha}$ chain expression in human basophilic KU812F cells. Ethanol extracts from root and stem of A. chiisanensis were tested for inhibitory effects of $Fc{\varepsilon}RI\;{\alpha}$ chain expression. The cell surface $Fc{\varepsilon}RI\;{\alpha}$ chain expression was examined by flow cytometric analysis. All of the extracts of A. chiisanensis reduced the cell surface $Fc{\varepsilon}RI\;{\alpha}$ chain expression. Furthermore, A. chiisanensis extracts caused a decrease in the level of $Fc{\varepsilon}RI\;{\alpha}$ chain mRNA level and $Fc{\varepsilon}RI-mediated$ histamine release. These results suggest that root and stem extracts of A. chiisanensis play an important role in anti-allergic activity via down-regulation of $Fc{\varepsilon}RI\;{\alpha}$ chain expression and decrease in release of inflammatory mediator such as histamine.

• The Journal of Pediatrics of Korean Medicine
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• v.22 no.1
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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.