• Korean Journal of Plant Resources
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• v.34 no.3
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• pp.203-215
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• 2021

Bok choy is one of Brassica vegetables widely consumed worldwide. Brassica vegetables have been reported to exert various pharmacological activities such as antioxidant, anti-cancer and cardioprotective activity. However, studies on immunostimulatory activity of bok choy sprout have not been conducted properly. Thus, in this study, we investigated in vitro immunostimulatory activity of bok choy sprout extract (BCS) using mouse macrophage RAW264.7 cells. Our results showed that BCS increased the production of immunomodulators such as NO, iNOS, IL-1β, IL-6, IL-12, TNF-α and MCP-1, and phagocytic activity in RAW264.7 cells. BCS activated MAPK, NF-κB and PI3K/AKT signaling pathways. However, BCS-mediated production of immunomodulators was dependent on JNK, NF-κB and PI3K/AKT signaling pathways. the mRNA expression of TLR2 were significantly increased by BCS, TLR2 inhibition by anti-TLR2 dramatically suppressed the production of immunomodulators by BCS. In addition, TLR2 inhibition by anti-TLR2 significantly reduced BCS-mediated phosphorylation level of AKT, JNK and NF-κB. From these results, BCS may have immunostimulatory activity via TLR2-MAPK, NF-κB and PI3K/AKT signaling pathways. Therefore, BCS expected to be used as a potential immune-enhancing agent.

• The Korea Journal of Herbology
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• v.29 no.3
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• pp.51-58
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• 2014

Objectives : In order to investigate the efficacy of BJBB on atopic dermatitis, various anti-inflammatory factors were studied. Methods : In-vitro, inflammatory mediators, such as MTT and nitric oxide were detected after the addition of LPS with or without BJBB in Raw 264.7 cells. In-vivo, in order to verify the effectiveness of BJBB in atopic dermatitis animal model, its role in inflammation factors and histological changes were observed in NC/Nga mice. Results : BJBB showed cell viability of 100% or higher in all concentration in Raw 264.7 cells. BJBB inhibited LPS-induced productions of inflammatory mediators nitric oxide in RAW 264.7cells. BJBB treated group showed significant decrease in the expression of IL-1b, IL-6 and TNF-a by 40%, 80% and 44% respectively. Also the group showed decrease in the transcription of IL-1b, IL-6 and TNF-a mRNA in spleen by 41%, 93% and 39% respectively. BJBB treated group showed significant decrease in WBC, neutrophil, lympocyte and monocytes immune cell ratio in blood by 54%, 63%, 57% and 86% respectively. BJBB treated group showed decrease in the expression of IgG by 39% respectively. Also, infiltration of adipocytes into skin was suppressed and the thickness of epidermis and dermis were relatively decreased in the BJBB treated group. Conclusion : BJBB has an anti-inflammatory effects in NC/Nga mouse. Thus, these results suggested a beneficial effect of BJBB in treatment with Atopic dermatitis and inflammatory.

• BMB Reports
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• v.40 no.4
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• pp.486-493
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• 2007

Atopic dermatitis (AD) is a chronic inflammatory skin disease and the pathogenesis of AD is associated with the release of various cytokines/chemokines due to activated $Th_2$ immune responses. Synthetic oligodeoxynucleotides (ODNs) containing unmethylated CpG dinucleotide in the context of particular base sequence (CpG motifs) are known to have the immunostimulatory activities in mice and to convert from Th2 to Th1 immune responses in AD. We aimed to investigate that CpG ODN, especially phosphodiester form, can stimulate the protective immunity in NC/Nga mice with AD. We isolated BMDCs from NC/Nga mice and then, cultured with GM-CSF and IL-4 for 6 days, and treated for 2 days by either phosphorothioate ODN or phosphodiester ODN. CpG ODN-treated DCs resulted in more production of IL-12. When CpG ODN-treated DCs were intravenously injected into the NC/Nga mice, the NC/Nga mice with CpG ODN-treated DCs showed significant improvement of AD symptoms and decrease of IgE level. Histopathologically, the NC/Nga mice skin with CpG ODN-treated DCs showed the decreased IL-4 and TARC expression comparing with non-injected mice. These results may suggest that phosphodiester CpG ODN-treated DCs might function as a potent adjuvant for AD in a mouse model.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.995-1000
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• 2004

This study was purposed to investigate the effect of Gamgung-tang(GGT) on immune responses induced by glucocorticoid in mice. GGT solution was treated by intraperitoneal injection for 7 days after glucocorticoid treatment(80㎎/㎏). And then B and T cell proliferation and cytolytic activity of natural killer(NK) cells were measured. There was 25% inhibition in B cell proliferation with treatment of glucocorticoid. However, B cell proliferation was not influenced by GGT treatment. T cell proliferation was also inhibited by 18.4% with treatment of glucocorticoid. On the other hand, T cell proliferation was increased dose-dependent manner in GGT treated group. Furthermore in purified T cell, the proliferation was furtherly increased than non-purified T cell. At concentration of 18㎎/mouse GGT, purified T cell proliferation was increase to above level of normal group. The cytotoxic activity of NK cell was decreased by 35.3% with treatment of glucocorticoid. In GGT treated group, the cytotoxic activity of NK cell was increased to the normal level. In purified NK cell, the cytolytic activity of NK cell was further increased than non-purifed NK cell. These results suggest that GGT may proliferate T cell that is suppressed by glucocorticoid, and activate NK cell activity.

• IMMUNE NETWORK
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• v.3 no.3
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• pp.248-254
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• 2003

Oral administration of antigen has long been used in the induction of immune tolerance in various animal models of autoimmune diseases including rheumatoid arthritis (RA). Alleveation of arthritogenic symptoms has been reported from RA patients who received oral administration of type II collagen (CII) without side effects, however its rather inconsistent therapeutic efficacy and variation among patients calls for more detailed investigation on the mechanism of oral tolerance to be settled as regular treatment for RA. In an attempt to understand the immunogenic processes underpinning tolerance induction by orally administered CII, we analyzed changes in the expression of costimulatory molecules and STAT/SOCS signaling messengers in the mouse model of collagen induced arthritis (CIA). We found thatin the spleen of CIA mice, that has been undergone repeated oral feeding of CII prior to the induction of arthritis, showed increased promortion of CTLA4 expressing lymphocytes than in the spleen of PBS fed control. On the other hand, cells expressing CD28 or ICOS were decreased in the spleen of tolerized mice. Tolerance induction by oral CII administration also enhanced the expression of STAT6 in both RNA and protein level, while not affecting the expression of STAT3. The expression of SOCS3, which hasbeen known to transmit STAT-mediated signals from Th2 type cytokines, remained unchanged in the spleen of tolerized mice. Interestingly transcript of SOCS1, which has been associated with Th1 related pathways, was only visible in the spleen of tolerized but not of control mice, suggesting that as in the case of IL-6 signaling, it may exert a feed back inhibition toward the Th1 type stimulation.

• Journal of Yeungnam Medical Science
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• v.30 no.1
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• pp.10-16
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• 2013

Background: Toll-like receptors (TLRs) are well-known pattern recognition receptors. Among the 13 TLRs, TLR2 is the most known receptor for immune response. It activates mitogen-activated protein kinases (MAPKs), which are counterbalanced by MAPK phosphatases [MKPs or dual-specificity phosphatases (DUSPs)]. However, the regulatory mechanism of DUSPs is still unclear. In this study, the effect of a TLR2 ligand (TLR2L, Pam3CSK4) on DUSP4 expression in Raw264.7 cells was demonstrated. Methods: A Raw264.7 mouse macrophage cell line was cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and 1% antibiotics (100 U/mL penicillin and 100 g/mL streptomycin) at $37^{\circ}C$ in 5% $CO_2$. TLR2L (Pam3CSK4)-mediated DUSP4 expressions were confirmed with RT-PCR and western blot analysis. In addition, the detection of reactive oxygen species (ROS) was measured with lucigenin assay. Results: Pam3CSK4 induced the expression of DUSP1, 2, 4, 5 and 16. The DUSP4 expression was also increased by TLR4 and 9 agonists (lipopolysaccharide and CpG ODN, respectively). Pam3CSK4 also induced ERK1/2 phosphorylation and ROS production, and the Pam3CSK4-induced DUSP4 expression was decreased by ERK1/2 (U0126) and ROS (DPI) inhibitors. U0126 suppressed the ROS production by Pam3CSK4. Conclusion: Pam3CSK4-mediated DUSP4 expression is regulated by ERK1/2 and ROS. This finding suggests the physiological importance of DUSP4 in TLR2-mediated immune response.

• IMMUNE NETWORK
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• v.6 no.3
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• pp.128-137
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• 2006

Background: Apoptosis is a physiologic phenomenon involved in development, elimination of damaged cells, and maintenance of cell homeostasis. Deregulation of apoptosis may cause diseases, such as cancers, immune diseases, and neurodegenerative disorders. The mouse myeloma cell P3-X63-Ag8.653 (v653) is an HGPRT deficient $(HGPRT^-)$ mutant strain. High dependency on de novo transcription and translation of aminopterin induced apoptosis of this cell seems to be an ideal experimental system for searching apoptosis-induced genes. Methods & Results: For searching apoptosis-related genes we carried out GE-array (dot blot), Affymetrix GeneChip analysis, Northern analysis and differential display-PCR techniques. The chip data were analyzed with three different programs. 66 genes were selected through Affymetrix GeneChip analyses. All genes selected were classified into 8 groups according to their known functions. They were Genes of 1) Cell growth/maintenance/death/enzyme, 2) Cell cycle, 3) Chaperone, 4) Cancer/disease-related genes, 5) Mitochondria, 6) Membrane protein/signal transduction, 7) Nuclear protein/nucleic acid binding/transcription binding and 8) Translation factor. Among these groups number of genes were the largest in the genes of cell growth/maintenance/death/enzyme. Expression signals of most of all groups were peaked at 3 hour of apoptosis except genes of Nuclear protein/nucleic acid binding/transcription factor which showed maximum signal at 1 hour. Conclusion: This study showed induction of wide range of proapoptotic factors which accelerate cell death at various stage of cell death. In addition apoptosis studied in this research can be classified as a type 2 which involves cytochrome c and caspase 9 especially in early stages of death. But It also has progressed to type 1 in late stage of the death process.

• Korean Journal of Veterinary Research
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• v.27 no.1
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• pp.53-60
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• 1987

This experiment was performed on mice to investigate the effects of an immunopotentiator, picibanil(PC), on the immune responses such as phagocytic activity of reticuloen-dothelial(RE) system, E rosette formation rate of splenic lmphocytes and morphological changes of lymph node tissue. Groups of mice were treated with a single(1KE/kg BW) or sequential(0.1, 0.25 and 0.5KE/kg BW for successive 3 days) intravenous injections of PC. PC treated and untreated control mice were sensitized with 50% sheep erythrocyte suspension(0.2ml/mouse) at 1, 3, 5, 7 and 10 days after PC treatment. Functional and morphological examinations were carried out 5 days after sensitization. The following results were obtained: The phagocytic activity of RE system and the weight of liver and spleen were increased significantly at 3rd, 5th and 7th day. The peripheral polymorphonuclear leukocyte and percent of lymphocyte and monocyte were slightly increased. The rates of E rosette formation of splenic lymphotytes, sequential PC treated groups were more increased at 3rd and 5th day in sequential PC treated groups than in single treated groups. Thereafter it returned gradually to the control level by the time of 10th day. Microscopically primary lymph follicles with indistinct germinal center (GC) were partially disrupted and the parafollicular areas were consisted of the pyroninophilic cells in control group. In PC treated group, the parafollicular areas were markedly proliferated and developments of secondary lymph follicles with enlarged and prominent GC were more pronounced in the sequential injected groups compared to single injected groups. These results indicate that PC affected not only parafollicular area of the T-cell area, but also GC of the B-cell area. It suggests that PC may potentiate both cell mediated immunity and humoral immunity.

• Journal of Sasang Constitutional Medicine
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• v.22 no.4
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• pp.77-84
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• 2010

1. Objectives Yanggyuksanhwa-tang for Soyangin was applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses on murine macrophage cell line (RAW 264.7) and ovalbumin/aluminium (OVA/Alum)-immunized mice. 2. Methods This study were carried out in nitric oxide (NO) synthesis on RAW 264.7 cells and cellular proliferation on mouse splenocytes. C57BL/6 mice were immunized intraperitonially with OVA/Alum (100 ${\mu}g$/200 ${\mu}g$) on day 1, 8, and 15. Yanggyuksanhwa-tang was administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results Yanggyuksanhwa-tang significantly enhanced cellular proliferation by LPS and Con A on splenocytes from OVA/Alum-immunized mice (p<.001). Yanggyuksanhwa-tang also significantly enhanced plasma OVA-specific IgG (p<.001), IgG1 (p<.001), and total IgM (p<.01) levels compared with the OVA/Alum group. 4. Conclusions These results suggested that Yanggyuksanhwa-tang for Soyangin could be used as immunopotent.

• Journal of Digital Convergence
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• v.14 no.11
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• pp.607-618
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• 2016

In order to clarify the Gosambaeksunpibokhap-bang(GBBB)'s therapeutic possibility on atopic dermatitis (AD), influences of GBBB on the changes of various immune-related factors and histological changes in NC/Nga mice were evaluated. Experimental results are as follows. Sign of recovery from AD was observed in GBBB treated group with naked eye test. The ratio of white blood cells, neutrophil, lymphocytes, monocyte in blood were decreased to 54%, 63%, 57% and 86% respectively in the GBBB group. IL-4, IL-5, IL-13, Histamine and IgE were significantly decreased to 40%, 80%, 62%, 61% and 57% respectively in the GBBB group. H&E staining showed thickness of epidermis and dermis were decreased by GBBB and inhibited the infiltration of lymphocytes. On the basis of these results, GBBB was confirmed that the possibility as an AD treatment applied externally to the skin. In the further study, immune control mechanism of GBBB will be demonstrated through the additional molecular biological research.