• Biomolecules & Therapeutics
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• v.24 no.5
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• pp.529-535
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• 2016

Atopic dermatitis (AD) results from gene and environment interactions that lead to a range of immunological abnormalities and breakdown of the skin barrier. Protease-activated receptor 2 (PAR2) belongs to a family of G-protein coupled receptors and is expressed in suprabasal layers of the epidermis. PAR2 is activated by both trypsin and a specific agonist peptide, SLIGKV-$NH_2$ and is involved in both epidermal permeability barrier homeostasis and epithelial inflammation. In this study, we investigated the effect of lobaric acid on inflammation, keratinocyte differentiation, and recovery of the skin barrier in hairless mice. Lobaric acid blocked trypsin-induced and SLIGKV-$NH_2$-induced PAR2 activation resulting in decreased mobilization of intracellular $Ca^{2+}$ in HaCaT keratinocytes. Lobaric acid reduced expression of interleukin-8 induced by SLIGKV-$NH_2$ and thymus and activation regulated chemokine (TARC) induced by tumor necrosis factor-a (TNF-${\alpha}$) and IFN-${\gamma}$ in HaCaT keratinocytes. Lobaric acid also blocked SLIGKV-$NH_2$-induced activation of ERK, which is a downstream signal of PAR2 in normal human keratinocytes (NHEKs). Treatment with SLIGKV-$NH_2$ downregulated expression of involucrin, a differentiation marker protein in HaCaT keratinocytes, and upregulated expression of involucrin, transglutamase1 and filaggrin in NHEKs. However, lobaric acid antagonized the effect of SLIGKV-$NH_2$ in HaCaT keratinocytes and NHEKs. Topical application of lobaric acid accelerated barrier recovery kinetics in a SKH-1 hairless mouse model. These results suggested that lobaric acid is a PAR2 antagonist and could be a possible therapeutic agent for atopic dermatitis.

• Journal of Haehwa Medicine
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• v.19 no.2
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• pp.119-137
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• 2011

In order to improve efficacy of oriental medicines and to study the application of fermented oriental medicine in clinicals, the efficacy of CYT and CYTBH on atopic dermatitis were evaluated. The results and conclusions are as follows. CYT and CYTBH significantly improved the atopic dermatitis symptoms in NC/Nga mice by naked eye evaluation and significantly decreased clinical index in both groups. CYT and CYTBH both decreased the cell numbers of CD3+, CD11b+Gr-1+ cells in dorsal skin. Of the cells, CYT significantly decreased CD11b+Gr-1+ cells whereas CYTBH significantly decreased all immune cells. CYT and CYTBH both decreased the production rate of IL-4 and IFN-${\gamma}$ activated by CD3/CD28. In the case of CYTBH, significant decrease in all cases was observed. CYT and CYTBH decreased the production rate of IL-5, IL-13 and IL-17 in serum. Significant decrease of IL-5 in the case of CYT and IL-5 and IL-13 in the case of CYTBH were observed. CYT and CYTBH significantly decreased transcription of IL-5 mRNA and IL-13 mRNA in skin. Significant decrease in IgG1 and IgE immunoglobulins in serum were oberved in both groups. Significant decrease was only observed in the case of CYTBH. Both CYT and CYTBH significantly decreased the secretion of histamine. Both CYT and CYTBH suppressed erythema, hemorrhage, edema, excoriation, erosion of skin tissues of NC/Nga mice resulting in the decrease of thickness of epidermis. Significant decrease of infiltration of obese cells was also observed. The results above indicated that both CYT and CYTBH had significant efficacy in the treatment of atopic dermatitis through immune modulation. Animal studies showed that CYTBH had superior activity than that of CYT suggesting further and continuous studies on the changes in ingredients or absorption improvement by fermentation should follow.

• Korean Journal of Veterinary Service
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• v.42 no.4
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• pp.257-264
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• 2019

Mastoparan V1 was used as adjuvant of formalin-inactivated Salmonella Gallinarum whole cells vaccine against fowl typhoid in a chicken model. The 75 brown nick chickens were equally divided into 5 groups, and all chickens of each group were immunized at 6 weeks of age (0 WPPI; weeks prime post immunization), and at 9 weeks of age (3 WPPI) (except group B). Group A chickens were intramuscularly (IM) inoculated with 500 uL of sterile phosphate-buffered saline (PBS), and group B chickens were subcutaneously immunized with 0.2 ml containing 5×10⁷ viable vaccine strain/bird. The chickens in groups C~E were IM inoculated with approximately 3×10⁹ cells/0.5 mL of formalin-inactivated the S. Gallinarum whole cells, approximately 3×10⁹ cells/0.5 mL of formalin-inactivated the S. Gallinarum whole cells with mastoparan V1 as adjuvant, and 0.5 mL of PBS, respectively. S. Gallinarum outer membrane proteins-specific serum IgG titers were considerably higher in groups B~D than in groups A and E. However, the levels of IFN-γ in groups B and D only than in groups A and E were significantly higher. Following oral challenge with virulent wild-type S. Gallinarum, no chicken in groups A (no challenge group) and B was dead, and only 30% of chickens in group D was dead. However, 70% of chickens in group C and all chickens in group E were dead after oral challenge. The results of this study demonstrated that IM immunization with approximately 3×10⁹ of the formalin-inactivated S. Gallinarum whole cells containing mastoparan V1 induced robust antibody and cell-mediated immune responses in chickens. The whole cells also conferred protection against infection with wild-type S. Gallinarum.

• 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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.4
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• pp.982-991
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• 2007

The present study was done to assess the suppressive effects of Haedongpi-san(HDPS), a traditional herbal medicine, on collagen induced arthritis (CIA) in mice and to examined it's effects on immune system. Oral administration of HDPS (200 or 400 mg/Kg) significantly suppressed the progression of CIA, which extend is comparable to that of methotrexate (MTX, 30 mg/Kg), a positive control. Histological examinations reveled that HDPS inhibited infiltration of inflammatory cells into affected paw joint, and bone erosion and cartilage destruction were greatly reduced compared with control. In paw joint, the number of CD3+ cells and CD11b+/Gr-1+ cells were greatly reduced by HDPS. The levels of pathologic cytokines including TNF-a and IL-6 were significantly decreased in the serum by oral treatment with HDPS. The levels of $IFN-{\gamma}$ in the culture supernatant of splenocyte stimulated with CD3/CD28 or collagen were dramatically decreased, while those of IL-4 was increased. Rheumatoid factors including IgG, IgM and collagen specific antibody were present much lower in the serum of HDPS treated mice than control. In peripheral blood mononuclear cells of HDPS treated mice, the percentage of CD3+, CD3+/CD69+, CD4+, CD4+/CD25+ cells were significantly decreased, while CD19+ cells were slightly increased compared with control. The absolute number of CD19+, CD3+, CD3+/CD69+, CD4+/CD25+, CD49b+ cell in spleen from HDPS treated mice were significantly decreased. The absolute number of CD3+, CD3+/CD69+, CD4+, CD4+/CD25+ CD8+, CD49b+, CD3+/CD49b+ cells in draining lymph node were significantly increased compared with control. Taken together, HDPS has suppressive effects on rheumatoid arthritis by modulating immune system, and has potential to use as an therapeutic for rheumatoid arthritis.

• Korean Journal of Veterinary Service
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• v.43 no.2
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• pp.89-97
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• 2020

This study was carried out to examine a novel inactivated Salmonella Typhimurium (S. Typhimurium) vaccine candidate for protection of mice against salmonellosis by immunization of BALB/c mice using various type adjuvant. The novel type-inactivated vaccine candidate was constructed by adding Chlorhexidine digluconate solution. BALB/c mice were divided into 6 groups of 15 mice apiece. The mice were intramuscularly (IM) primed at 6 weeks of age and were IM boosted 8 weeks of age. Groups A and B mice were injected with sterile phosphate-buffered saline as controls; group C mice were inoculated with 5×10⁸ cells/100 µL of formalin-inactivated S. Typhimurium cells and adjuvant ISA70; groups D~F mice were immunized with 5×10⁸ cells/100 µL of the inactivated vaccine candidate and adjuvant ISA70, adjuvant IMS1313 and adjuvant IMS1313 containing 30 ㎍/mL of GI24, respectively. All mice (except group A mice) were orally challenged with a virulent S. Typhimurium strain at 10 weeks of age. Mice from groups C-F had significantly increased IgG levels compared to control groups (A-B) mice. The levels of splenocyte IFN-γ and IL-4 in mice of all groups were measured by ELISA, resulting in increased immunity in group F mice compared to those of groups A-E mice. These data suggested that systemic and cell-mediated immune responses were highly induced by IM immunization with the vaccine candidate and adjuvant IMS1313 containing GI24. Furthermore, clinical signs such as death were observed in only 20% of group F mice after virulent Salmonella strain challenge, however, groups B and C (100%), and groups D and E (60%) mice died. This data suggested that mice immunized by intramuscular prime and booster with this vaccine candidate and adjuvant IMS1313 containing GI24 effectively protected mice from salmonellosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.3
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• pp.700-704
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• 2007

In the recent, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the role of the extract of Anethi Fructus in the expression of inflammatory mediators, surface molecule, and related receptors in vitro. In murine macrophage RAW 264.7 cells and peritoneal macrophages of C57BL/6N mice, water extract of Anethi Fructus increased the production of secretary tumor necrosis factor (TNF)-a and Nitric oxide (NO), and the expression level of CD14, LPS co-receptor and CD86, co-stimulatory molecule compared to negative natural extract ex vivo. The water extract of Anethi Fructus increased the production of interferon (IFN)-g from splenocytes. Also, water extract of Anethi Fructus increased ConA-induced cell proliferation. These results suggest that water extract of Anethi Fructus may enhance the immune response through immune modulation of macrophage and lymphocytes.

• The Korea Journal of Herbology
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• v.34 no.2
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• pp.41-47
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• 2019

Objectives : A traditional herbal prescription, Kyung-Ok-Ko (KOK), has long been used in oriental medicine as an invigorant for age-related diseases, such as amnesia and stroke. However, the beneficial value of KOK for immune responses is largely unknown. Based on the above mentioned effects of KOK, other previous reports, and its use in traditional medicine, we hypothesized that KOK displays beneficial effects against methotrexate (MTX)-induced immune suppression. Methods : We investigated the effects of KOK (0.6 g/kg/day, oral (p.o.)) on deteriorated immunity caused by MTX (2 mg/kg/day, p.o.) in an immune suppression mouse model. MTX was fed to mice once a day for 7 days. After the immune responses of the mice deteriorated by MTX treatment, KOK in water was fed to the mice once a day for 14 days. We then measured the expression levels of various cytokines, such as T helper cell (Th1, Th2) cytokines, and the number of immune cells, such as spleen T cells, B cells, and macrophages. Results : The data showed that MTX decreased Th1 profiles (interferon $(IFN)-{\gamma}$, interleukin (IL)-2, IL-12) and the number of immune cells, and increased Th2 profiles (IL-4, IL-5, IL-13), which were normalized significantly by post-administration of KOK. However, there was no significant difference in body-weight gain between MTX- and KOK-treated mice. Conclusion : These results indicate that KOK has immune-enhancing functions and reduces immunotoxicity of MTX, suggesting that supplementation with KOK will improve immune responses clinically and be useful for the prevention of immune-related diseases.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.166-182
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• 2022

Deer antler velvet is widely used in traditional medicine for its anti-aging, antioxidant, and immunity-enhancing effects. However, few studies have reported on the discovery of probiotic strains for deer antler fermentation to increase functional ingredient absorption. This study evaluated the ability of probiotic lactic acid bacteria to enhance the concentrations of bioactive molecules (e.g., sialic acid and gamma-aminobutyric acid [GABA]) in extracts of deer antler velvet. Seventeen strains of Lactobacillus spp. that were isolated from kimchi and infant feces, including L. sakei, L. rhamnosus, L. brevis, and L. plantarum, and those that improved the life span of Caenorhabditis elegans were selected for evaluation. Of the 17 strains, 2 (L. rhamnosus LFR20-004 and L. sakei LFR20-007) were selected based on data showing that these strains increased both the sialic acid and GABA contents of deer antler extract after fermentation for 2 d and significantly improved the life span of C. elegans. Co-fermentation with both strains further increased the concentrations of sialic acid, GABA, and metabolites such as short-chain fatty acids and amino acids. We evaluated the biological effects of the fermented antler velvet (FAV) on the antibacterial immune response in C. elegans by assessing worm survival after pathogen infection. The survival of the C. elegans conditioned with FAV for 24h was significantly higher compared with that of the control worm group fed only normal feed (non-pathogenic E. coli OP50) exposed to E. coli O157:H7, Salmonella typhi, and Listeria monocytogenes. To evaluate the protective effects of FAV on immune response, cyclophosphamide (Cy), an immune-suppressing agent was treated to in vitro and in vivo. We found that FAV significantly restored viability of mice splenocytes and immune promoting-related cytokines (interleukin [IL]-6, IL-10, inducible nitric oxide synthase [iNOS], interferon [IFN]-γ, and tumor necrosis factor [TNF]-α) were activated compared to non-fermented deer antlers. This finding indicated the protective effect of FAV against Cy-induced cell death and immunosuppressed mice. Taken together, our study suggests that immune-promoting antler velvet can be produced through fermentation using L. rhamnosus LFR20-004 and L. sakei LFR20-007.

• The Korean Journal of Food And Nutrition
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• v.36 no.1
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• pp.6-16
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• 2023

To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.