• 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.

• 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.

• Fisheries and Aquatic Sciences
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• 제27권10호
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• pp.699-707
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• 2024

Codium fragile (CF) is a green seaweed that has been used in folk medicine to treat urinary tract disorders, intestinal bacteriosis, and dropsy. The CF is also used primarly as a seasoning to enhance the flavor of Kimchi, and in some areas it is consumed as a side dish. The CF has reported to have anti-coagulant, thrombolytic, anti-inflammatory, anti-oxidant activities. The purpose of this study was to analyze the immune-enhancing effect of aqueous extract of C. fragile (AECF) using RAW264.7 macrophages and splenocytes. The AECF induced the production of nitrite oxide and inducible nitric oxide synthase at 1 mg/mL without cytotoxicity, and also gradually increased the expression of pro-inflammatory cytokines (tumor necrosis factor-alpha and interleukin-6), determined by griess reagent, western blot, and Enzyme-linked immunosorbent assay. In addition, AECF significantly induced the expression of immune-related enzymes, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, and monocyte chemoattracted protein-1, which increase the production of immune cells, determined by mouse cytokine array, as determined by cytokine arrray. The AECF induced phosphorylation of mitogen-activated protein kinases and translocation of nuclear factor kappa B p65 subunit from cytosol to the nucleus. Our findings indicate that C. fragile may have potential as a functional ingredinet for developing immune-enhancing supplement that properly maintiain the body's defense system.

• Toxicological Research
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• 제31권1호
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• pp.49-59
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• 2015

Eye is a highly vascularised organ. There are chances that a foreign substance can enter the systemic circulation through the eye and cause oxidative stress and evoke immune response. Here the eyes of rabbits were exposed, for a period of 7 days, to 5 known ocular irritants: Cetyl pyridinium chloride (CPC), sodium salicylate (SS), imidazole (IMI), acetaminophen (ACT) and nicotinamide (NIC). The eyes were scored according to the draize scoring. Blood collected from the treated rabbit were analyzed for haematological and biochemical parameters. After sacrifice, histological analysis of the eye and analysis of pro-inflammatory biomarkers ($IL-1{\alpha}$, $IL-1{\beta}$, IL-8 and $TNF-{\alpha}$) in the cornea using ELISA was carried out. Spleen was collected and the proliferation capacities of spleenocytes were analyzed. Liver and brain were collected and assessed for oxidative stress. The eye irritation potential of the chemicals was evident from the redness and swelling of the conjunctiva and cornea. Histopathological analysis and ELISA assay showed signs of inflammation in the eye. However, the haematological and biochemical parameters showed no change. Spleenocyte proliferations showed only slight alterations which were not significant. Also oxidative stress in the brain and liver were negligible. In conclusion, chemicals which cause ocular irritation and inflammation did not show any systemic side-effects in the present scenario.

• 대한한방소아과학회지
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• 제35권1호
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• pp.76-103
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• 2021

Objectives This study was designed to examine the effect of Gleditsiae Fructus n-hexane (GSF_Hx) on two different groups (on the LPS-induced activation of Raw264.7 cells in vitro, and on the DNCB-induced activation of atopic dermatitis NC/Nga Tnd mice in vivo) to find index components and active components of Gleditsiae Fructus. Methods GSF_Hx was analyzed by HPLC profiling and confirmed echinocystic acid (EA), oleanolic acid (OA) as index components of Gleditsiae Fructus. Using GSF_Hx, EA, OA, we investigated IL-6, TNF-α, NO production by ELISA analysis and evaluated manifestations of MAPKs transcription factors and NF-κB p65 translocation by western blotting. During In vivo study, atopic dermatitis was induced on NC/Nga Tnd mice by DNCB and administered GSF_Hx, EA, OA orally, and checked skin lesions and measured skin clinical score. Serum IgE level, Th1 and Th2 cytokines secretion and modulating molecular mediators and immune cells in the spleenocyte culture supernatant, PBMCs, ALN and dorsal skin were also measured by real-time PCR. Then, skin rash was evaluated and mast cell distribution was verified by H&E and toluidine blue staining on dorsal skin. Results It is possible that GSF_Hx, EA and OA reduce inflammation and allergic response of atopic dermatitis by suppressing Th1 and Th2 cytokines secretion and modulating molecular mediators and immune cells. They also had moisturizing effect by raising vitality of ceramide in dorsal skin of atopic dermatitis NC/Nga Tnd mice. However, EA particularly had better overall activity data than OA, that EA could be a more effective active component of Gleditsiae Fructus than OA. Conclusions Based on the inflammatory reduction property with moisturizing effect, GSF_Hx may play a role in effective treatment for atopic dermatitis.

• 대한한방소아과학회지
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• 제29권4호
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• pp.39-66
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• 2015

Objectives The purpose of this study is to investigate the effects of Sesamum indicum extracted (SEI) on atopic dermatitis in an in-vitro and in-vivo experiment using a MC/9 murine mast cells and a NC/Nga mouse. Methods In-vitro experiment, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression were evaluated by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA and manifestations of NFAT-1, NFAT-2, c-jun, c-fos, NF-${\kappa}B$ p65 transcription factors by western blotting. In-vivo experiment, we measured WBC, Eosinophil, Neutrophil, and serum IL-5, IL-13 in NC/Nga atopic dermatitis mouse, IL-5, IL-13, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant by ELISA, the absolute cell numbers of CD4+, CD8+, +Gr-1+CD11b, B220+CD23+ in the axillary lymph node (ALN), peripheral blood mononuclear cells (PBMCs) and dorsal skin tissue, IL-5, IL-13 by Real-time PCR, the distribution of tissue inflammation and cellular infiltration by H&E and toluidine blue. Results SEI decreased IL-4, IL-5, IL-6, IL-13, GM-CSF, TNF-${\alpha}$ mRNA expression, IL-13, MIP-$1{\alpha}$ production and the expression of transcription factors including NFAT-1, c-jun, NF-${\kappa}B$ p65 in MC/9 murine mast cells. SEI orally administration decreased cell number of WBC, Eosinophil, the level of serum IgE, total cell number of ALN and dorsal skin tissue, absolute cell number of CD4+, CD8+, B220+CD23+ in the ALN. SEI orally administration also increased absolute cell number of CD8+/CD3+ and decreased Gr-1+/CD11b+ in PBMCs, decreased CD4+ in dorsal skin tissue, inhibited IL-5, IL-13 mRNA expression. Infiltration levels of inflammatory immune cells, mast cells and thickness of epidermis decreased in dorsal skin tissue. Conclusions SEI can regulate allergic inflammatory response suppressed the gene expression and production of cytokines that mediate allergic reactions, and will be able to be effectively utilized in the treatment of atopic dermatitis future.

• 대한한방소아과학회지
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• 제25권1호
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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.

• 대한본초학회지
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• 제32권4호
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• pp.1-8
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• 2017

Objectives : Silkworm is known as immunomodulatory substances and contain various bioactive compounds such as serine, tyrosine and alanine. The aim of this study was to investigated the immunopotentiating activity of combine extract that silkworm and food materials (Eucommia ulmoides, Angelica gigas, Acanthopanax, Allium hookeri, Cinnamomum cassia, Liriope platyphylla, Curcuma longa, Achyranthes japonica, Alpinia oxyphylla, Adenophora triphylla). Methods : Among 10 kinds of food materials, to select food materials with the effect of enhancing the immune function mouse splenocyte proliferation ability was measured by 3-[4,5-dimethylthiazol-2-y]-2,5-diphenyl terazolium bromide (MTT) assay. Then, combine extract of silkworm and food materials were evaluated that mouse splenocyte proliferation ability by EZ-cytox cell viability assay. Morever, cytokines production such as IL-2, IL-4, IL10, IL12, $IFN-{\gamma}$ on mouse T lymphocyte stimulated with concanavalin A (ConA) was measured. Results : Eucommia ulmoides, Acanthopanax, Allium hookeri, Cinnamomum cassia, Liriope platyphylla has high proliferation ability of mouse splenocyte compared with Curcuma longa, Achyranthes japonica, Alpinia oxyphylla, Adenophora triphylla. The silkworm and food material combined extract has a relatively high proliferation ability of mouse splenocyte proliferation when the silkworm and food materials are used as a single material. In particularly, combined extract of silkworm and Cinnamomum cassia was stimulate cytokine production on T lymphocyte such as IL12, $IFN-{\gamma}$. Combined extract of silkworm and Liriope platyphylla was stimulate cytokine production on T lymphocyte such as IL2, IL4, IL10. Conclusion : In conclusion, the combined extract of the silkworm and Cinnamomum cassia or Liriope platyphylla may enhance immune function by regulating mouse splenocyte proliferation and stimulating cytokine production.

• 한국식품과학회지
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• 제40권1호
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• pp.82-87
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• 2008

아토피 피부염에서 유산균의 유익성에 대해서는 이미 많이 알려져 있고 우리나라 전통식품인 김치유산균의 유익성에 대한 연구도 많이 진행되었으나 아토피 피부염에서 김치 유산균의 효과에 관한 연구는 아직 미비한 실정이다. 본 연구에서는 아토피 동물 모델인 NC/Nga mice에 김치유산균에서 추출한 유산균(L.plantarum K8)의 파쇄물과 유산균 및 ${\gamma}$-리놀렌산 복합체(L.rhamnosus GG 생균, B. lactis Bb-12Lb 생균, L. plantarum K8생균, L. plantarum K8 파쇄물, ${\gamma}$-리놀렌산)를 경구 투여한 후 혈청 면역지표(IgE, IL-4, IL-5, IFN-${\gamma}$)와 비장 배양액에서 IgE 농도를 측정하였다. 실험동물의 체중 및 사료섭취량은 사육기간이 지남에 따라 증가하는 경향을 보였으며 각 군 간에 유의적인 차이는 관찰되지 않았다. 실험 시작 시 혈청 IgE 농도는 각 군 간에 차이가 나타나지 않았으나 실험 전 기간 4주 동안 유산균 파쇄물(DPF), 유산균 및 ${\gamma}$-리놀렌산 복합체(DPOC), 항히스타민(DAH)을 섭취한 군에서 증류수(DDW)만을 섭취군에 비하여 유의적으로 낮게 나타났다. 혈청 IFN-${\gamma}$ 농도는 각 군에서 유의적인 차이가 나타나지 않았으나 혈청 IL-4 농도는 유산균 및 ${\gamma}$-리놀렌산 복합체를 섭취한 군(DPOC)에서 대조군에 비하여 유의적으로 낮게 나타났다. 혈청 IL-5 농도는 유산균 파쇄물(DPF), 유산균 및 ${\gamma}$-리놀렌산 복합체(DPOC), 항히스타민(DAH)을 섭취한 군에서 모두 대조군에 비하여 유의적으로 낮았다. 비장 임파구 배양 상층액의 IgE 농도는 각 군에서 군 간 유의적인 차이가 나타나지 않았다. 결론적으로, 아토피 동물모델 NC/Nga mice에 유산균 파쇄물(DPF)과 유산균 및 감마리놀렌산 복합체(DPOC)의 경구투여는 혈청 IgE, IL-4, IL-5를 감소시키는 것으로 나타났으며, 김치로부터 추출한 프로바이오틱스를 아토피 환자에게 섭취시켰을 때 IgE, IL-4, IL-5 감소와 같은 긍정적인 효과를 볼 수 있을 것으로 기대한다. 여러 종류의 프로바이오틱스는 아토피의 증상완화 또는 면역 체계에 미치는 영향과 정확한 메카니즘을 밝히기 위하여 과학적인 연구가 필요한 것으로 사료된다.