• 대한본초학회지
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• 제28권6호
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• pp.15-23
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• 2013

Objectives : To clarify the possible effects of Sinapis Semen and Raphani Semen on the development of pulmonary eosinophilic inflammation in a asthmatic mouse model. Methods : BALBav/c mice were sensitized to OVA followed intratracheally and by aerosol allergene challenges. We investigated the effect of Sinapis Semen and Raphani Semen on airway hyperresponsiveness, eosinophiic infitratio, immune cell phenotype, The2 cytokine product, and OVA-spedific IgE production. Results : Total lung cells, eosinophils, and lung leukocytes, OVA specific IgE levels, and Th 2cytokine levels such as IL-5, IL-13, IL-17, TNF-alpha, and eotaxin in BALF were reduced compared with those of OVA sensitized asthma mice (control). The absolute numbers of $CD3^+$, $CD3^+/CD69^+$, $CD3^-/CCR3^+$, $CD4^+$, $CD8^+$, $Gr-1^+/CD11b^+$, $B220^+/CD22^+$, $B220^+/IgE^+$ cells in lung tissiues significantly reduced compared to those of control. Specially total lung cells in BALF and the absolute number of $CD3^+/CD69^+$ and, $B220^+/IgE^+$ cells in lung tissiue effectively reduced in Sinapis Semen plus Raphani Semen compared to those of Sinapis Semen and Raphani Semen. Conclusions : These results indicate that Sinapis Semen plus Raphani Semen has deep inhibitory effects on airway inflammation and hyperresponsiveness in asmatic mouse model and also has effect of suppression of IL-5, IL-13, IL-17, OVA specific IgE production in BALF. The results verified that Sinapis Semen, Raphani Semen, and Sinapis Semen plus Raphani Semen could act as a immunomodulator which possess anti-inflammatory and anti-asthmatic property by modulating the relationship of Th1/Th2 cytokine imbalance.

• 혜화의학회지
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• 제16권2호
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• pp.171-190
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• 2007

Atopic dermatitis is a chronic inflammatory skin disease characterized by pruritic and erythromatous skin lesions. In this study we examined the suppressive effects of SJJY on der f induced atopic dermatitis in NC/Nga mic, and concluded as follows: Oral administration of SJJY significantly decreased the severity score in the skin lesions at the dosage of 6.6 mg/25g/day for 8 weeks. SJJY significantly suppressed the infiltration of inflammatory cells into skin compared with control, and decreased the expression of CD4, CD8, CD20 and CCR3 in the skin lesions. SJJY significantly decreased the level of IgE in the serum compared with control, and the levels of IgM, IgG2a and IgG2b were also decreased. SJJY significantly decreased the levels of IL-6, but not TNF-a, in the serum compared with control. The levels of IFN-$\gamma$ was significantly increased in the supernatant of CD3/CD28 activated cultured splenocytes from the SJJY treated mice. The levels of IL-4 and IFN-$\gamma$ in the supernatants was much less in the der f activated splenocytes from SJJY treated mice than control. SJJY significantly increased the total number of cells in lymph node, while decreased the total number of skin compared with control. SJJY increased the number of CD3+ and CD4+ cell compared with control, while decreased the number of CD4+/CD25+ and CCR3+ cells in the PBMC. SJJY increased the number of CD3+, CD4+, CD8+, CD4+/CD25+, NKT+, CD3+/CD69+ cells compared with control, while decreased the number of B220+/IgE+, B220+/CD23+ cells in the lymph node. SJJY significantly decreased the number of CD3+/CD69+, CCR3+, B220+/IgE+, CD11b+/Gr-1+ compared with control in the skin lesions. Taken together, these results suggested that SJJY has suppressive effects on atopic dermatitis by the regulation of immune system and has potential as a therapeutics for atopic dermatitis. Further studies on molecular mechanisms on immune regulation are needed.

• 동의생리병리학회지
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• 제21권1호
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• pp.39-49
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• 2007

Rheumatoid arthritis is an autoimmune disease involving multiple joint. In order to access the suppressive effects of JTT on rheumatoid arthritis and it's effects on immune system we investigated whether JTT could suppress the disease progression of collagen-induced arthritis. DBA/1 mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with DW, JTT (200 or 400 mg/kg) or methotrexate (MTX, 30 mg/kg) as a positive control. Oral administration of JTT significantly suppressed the progression of CIA, which extend is comparable to that of MTX. Histological examination reveled that JTT inhibited infiltration of inflammatory cells into affected paw joint and bone erosion and cartilage destruction were greatly reduced compared with control. Total cell number of spleen, lymph node and peripheral blood were significantly reduced. The absolute number of CD19$^+$, CD3$^+$/CD69$^+$, CD4$^+$/CD25$^+$ cell in spleen from JTT treated mice were significantly decreased. The absolute number of CD19$^+$, CD3$^+$, CD3$^+$/CD69$^+$, CD4$^+$, CD4$^+$/CD25$^+$ CD8$^+$, CD49b, CD3/CD49b cells in draining lymph node were significantly increased compared with control. In peripheral blood mononuclear cells of JTT treated mice, the absolute number of CD4$^+$, CD4$^+$/CD25$^+$, CD3$^+$/CD69$^+$ cells were significantly decreased compared with control, while that of CD49b$^+$ was slightly increased. Infiltration of CD3$^+$ cells and CD11b$^+$/Gr-1$^+$ cells into paw joint was significantly reduced in JTT treated mice. The levels of pathologic cytokines including TNF-a and IL-6 in serum were significantly decreased by oral treatment with JTT The levels of IFN-g in the culture supernatant of splenocyte stimulated with CD3$^+$/CD28$^+$ or collagen were dramatically decreased, while the levels of IL-4 was increased under CD3$^+$/CD28$^+$ or collagen stimulation. Rheumatoid factors including IgG, IgM and collagen specific antibody were present much lower in the serum of JTT treated mice than control. Taken together, JTT has suppressive effects on rheumatoid arthritis by modulating immune system, and has potential to use anti-rheumatic arthritic agent in human.

• 동의생리병리학회지
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• 제21권4호
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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.

• Journal of Acupuncture Research
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• 제22권6호
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• pp.1-15
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• 2005

Objectives : The aim of this study was to investigate the effect of Peucedani Radix herbal acupuncture(PR-HA) at St36(joksamni) on ovalbumin-induced asthma in mice. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks(once a week) C57BL/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (3times a week). One of the two experimental groups was just treated with needle-prick on St36 and the other group was treated with 1% concentrations of PR-HAS at St36, for the later 8 weeks (3times /week). Results : 1. The weight and total cells of lung of the mice group treated with PR-HA decreased significantly compared with those of control group. 2. Total Leukocytes and Eosinophils in BALF of the mice group treated with PR-HA decreased significantly compared with those of control group. 3. Eosinophils in BALF of the mice group treated with PR-HA in Photomicrographs decreased significantly compared with those of control group. 4. According to Histological analysis of lung sections, it decreased significantly adhension of collagen in PR-HA than those of control group 5. The concentration of IgE, IL-4, IL-5, in BALF and IL-4, IL-5, Il-13 in serum of the mice group treated with PR-HA decreased significantly compared with that of control group. 6. The number of Gr-1+/CD11b+, CD11b+, CD3-/CCR3+, CD4+, CD3e+/CD69+ , CD23+B220+ cells in the lungs of the mice group treated with PR-HA decreased significantly compared with those of control group. 7. The cytokine's manifestation of mRNA of the mice group treated with PR-HA with RT-PCR decreased significantly compared with that of control group. Conclusion : We conclude that PR-HA is effective on OVA-induced asthma of C57BL/6 mouse.

• 혜화의학회지
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• 제19권2호
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• pp.101-118
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• 2011

Herbal medicine has a high body absorption rate when it ferments. Biological and clinical research on the fermented herb gradually increases because it has effective materials for the treatment of a disease and it is a little bitter. In this study, we investigated the effect of fermented Baedokhwanbalhyobang (BDHBH) on attenuation of the development of atopic dermatitis in NC/Nga mice by evaluating the cytokine level in serum, the mRNA expression of cytokine and histological alteration of the skin, and the skin severity. We have come to the following conclusion. BDHBH led to a significant decrease in the skin severity score (63.1%) as compared to the control group. CD4+/CD45+, CD4+, B220+/CD23+, and CD11b+/Gr-1+cells of peripheral mononuclear cells (PBMCs) in the BDHBH-treated group were decreased to 6.7%, 31.1%, 22.4%, 36.6%, respectively. CD3+and CD11b+/Gr-1+immune cells in dorsal skin of the BDHBH-treated group were decreased to 52.9% and 28.0%. The levels of IL-5 and IL-13 in serum of the BDHBH-treated group were inhibited to 18.8% and 5.1%. The mRNA expressions of IL-5 and IL-13 in dorsal skin were also decreased to 30.6% and 27.8% after the treatment of BDHBH. BDHBH inhibited the proliferation and differentiation of eosinophils. In histological examination, BDHBH decreased the thickness of epidermis and dermis, and infilatration of mast cells as compared to the control group. These results indicate that BDHBH inhibits the pathogenic development of atopic dermatitis in NC/Nga mice. These results may indicate that BDHBH attenuates the development of atopic dermatitis-like lesions by lowering immune cells and inflammatory cytokine levels, and that it is valuable in drug development for the treatment of atopic dermatitis. Further experiments on the components of BDHBH will be needed to better understand the effect of a fermented herb as compared to a herb.

• 대한한방소아과학회지
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• 제28권4호
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• pp.1-29
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• 2014

Objectives Dictamni Radicis Cortex extracts (DRC) has been known to suppress allergic reaction, however the cellular target of DRC and its mode of action remain unclear. The purpose of this study is to investigate the effects of Dictamni Radicis Cortex extracts on DNCB induced atopic dermatitis-like skin lesions of NC/Nga mouse. Methods This study was designed to investigate the effects of DRC extract in the DNP-IgE-induced activation of MC/9 murine mast cell lines in vitro and in the DNCB-induced activation of NC/Nga mouse in vivo. For this investigation, We examined IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA analysis and manifestations of NFAT1, NFAT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting in vitro. Then, we examined WBC, eosinophil and neutrophil in NC/Nga mouse, IL-5, IL-13 in serum, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $^+Gr-1^+CD11b$, $B220^+CD23^+$ in the ALN, PBMCs and dorsal skin, IL-5, IL-13 in the dorsal skin by Real-time PCR and the distribution of mast cells by H&E and toluidine blue. Results In vitro the mRNA expression of IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$, GM-CSF and IL-13, MIP-$1{\alpha}$ production by ELISA analysis were completely abolished by DRC and the western blot analysis decreased the expression of mast cell-specific transcription factors including NFAT-1, NF-${\kappa}B$ p65. In vivo DRC oral adminstration also decreased the counts of WBC, eosinophils and inflammatory cytokines such as IL-13 and IgE in the serum. DRC oral adminstration elevated IL-4 level in the spleenocyte culture supernatant. DRC oral adminstration decreased total ALN cells, total skin cells, cell numbers of $CD4^+$, $B220^+CD23^+$ in the ALN, $^+Gr-1^+CD11b$ in the PBMCs and $CD4^+$, $CD8^+$ in the dorsal skin. The mRNA expression of IL-5, IL-13, thickness of epidermis, inflammation immune cells and mast cells were abolished by DRC in the dorsal skin. Conclusions Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mouse were much improved by DRC oral adminstration. These results, therefore, suggest that DRC can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis induced in NC/Nga mouse, and may play an important role in recovering AD symptoms.

• 대한한의학회지
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• 제31권4호
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• pp.63-78
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• 2010

Objectives: This study was carried out to know the effects of Gwan-Jul-Bang-7 (hereafter referred to GJB-7) on the inhibition of arthritis induced by collagen on the mouse. Methods: To assess the effects of GJB-7 on mouse with arthritis induced by collagen II, we conducted several experiments such as analysis of cytotoxicity, hepatotoxicity, arthritis index, total cell number of draining lymph nodes and paw joints, value of immunocyte in PBMC (peripheral blood mononuclear cell), DLN (draining lymph node) and paw joint, measurement of cytokine and anti-collagen II, observation of the histological changes of joint. Results: 1. Cytotoxicity against HFC (human fibroblast cells) was not observed in any concentration and hepatotoxicity was not observed in the GJB-7 treated group. 2. The incidence of arthritis significantly decreased. 3. Total cell number of draining lymph nodes significantly increased and total cell number of paw joints significantly decreased. 4. The percentage of $CD8^+$ cells in PBMC (peripheral blood mononuclear cell) significantly increased. The percentage of $CD3^+/CD69^+$, and $CD3^+/CD49b^+$ cells in PBMC significantly decreased. 5. The percentage of $CD19^+,\;CD3^+$, and $CD4^+/CD25^+$ cells in DLN (draining lymph nodes) significantly increased. The percentage of $B220^+/CD23^+$ cells in DLN significantly decreased. 6. The percentage of $CD3^+,\;CD4^+$, and $CD11b^+/Gr-1^+$ cells in paw joints significantly decreased. 7. The production of TNF-$\alpha$, IL-6, and MCP-1 significantly decreased. 8. Anti-collagen II in serum significantly decreased. 9. With the hematoxylin and eosin stain, inflammation of joint decreased. Under Masson's trichrome stain, the cartilage destruction and synovial cell proliferation and the expression of collagen fibers decreased. Conclusions: Comparison of the results for this study showed that GJB-7 had immunomodulatory effects. So we expect that GJB-7 could be used as an effective drug for not only rheumatoid arthritis but also another auto-immune diseases.

• BMB Reports
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• 제45권9호
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• pp.538-543
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• 2012

We evaluated the expression of the costimulatory molecules CD80 and CD83 and major histocompatibility (MHC) class II induced by 2,4-dinitrofluorobenzene (DNFB) in the RAW 264.7 macrophage cell line. In contrast to the previously reported effect of DNFB on dendritic cells, CD86 expression did not change. Furthermore, we observed that the CD83 expression level transiently increased and then decreased. Induction of CD80 and MHC class II molecule expression and a decrease in CD83 expression by DNFB in vitro were also confirmed in splenocytes of BALB/c and NC/Nga mice. However, DNFB did not influence CD83 expression in peritoneal $CD11b^+$ cells from BALB/c or NC/Nga mice. Detailed in vivo experiments and further studies on the possible contribution of $CD11b^+$ cells to induce atopic dermatitis (AD) would be helpful to attain a better understanding of AD pathogenesis.

• 대한한방소아과학회지
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• 제30권3호
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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.