• 한국식품영양과학회지
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• 제33권7호
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• pp.1085-1091
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• 2004

먼저 OVA항원에 대해 특이적으로 증식반응을 나타내는 T세포주를 수립하였고, 수림된 세포주는 세포 표면 단백질이 CD4$^{＋}$CD8$^{-}$이며 IL-2와 IFN-${\gamma}$를 분비하는 Type I에 속하는 보조 T세포(Thl)인 것을 확인하였다. 보중익기탕의 total 분획은 OVA항원에 대해 특이적으로 반응하는 Thl세포의 증식반응을 증가시키는 효과를 나타내지 않았으며 고농도에서 오히려 증식반응을 억제하였다. 그러나, 보중익기탕의 polysaccharide 분획은 전반적 인 농도에서 T세포의 증식반응을 유의하게 증가시키는 효과가 있는 것으로 나타났다. 보중익기탕의 polysaccharide 분획을 첨가하였을 때 T세포의 IL-2 분비량은 대조군보다 약간 적었지만, IFN-${\gamma}$ 분비량은 대조군보다 증가하였다. 그리고, 분비된 IL-2와 결합하는 T세포의 IL-3 수용체 발현양도 증가하는 것으로 나타났다. 또한, 항원제시세포의 MHC class II의 발현양도 증가시켰다. 이상의 결과로 보중익기탕의 polysaccharide분획은 T세포의 IL-2수용체 발현양을 증가시키고, 항원제시세포의 MHC classs II의 발현양을 증가시켜서 T세포의 증식반응을 증가시키는것으로 생각된다 그리고 보중익기탕이 생체 면역반응에 미치는 보다 정확한 효과를 평가하기 위해서는 직접 살아있는 실험동물에 투여하는 in vivo 실험이 필요하다.

• 대한본초학회지
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• 제32권5호
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• pp.27-38
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• 2017

Objective : In the present study, we examined whether Canavalia gladiata D.C. (CG) and Arctium lappa L., Redix (AL) mixture (CGAL), their components, lupeol and chicoric acid, regulate immune system and suppress the tumor in vitro and in vivo. Methods : LPS-induced reactive oxygen species (ROS) and nitric oxide (NO) were measured after treatment with CG extract (CGE), CGAL, lupeol, chicoric acid and lupeol and chicoric acid mixture (lupeol+CA) in Raw264.7 cell. To determine the effect of CGE on immune responses, immune cell population and IgG production were assessed in mice. To investigate the effect of CGAL and their component on anti-tumor activity, tumor volume and weight were measured, cell cycles and immune cell population were analyzed in MC38 injected tumor bearing mice. Also, NK cell activity was determined in splenocyte isolated from tumor bearing mice. Results : CGE, CGAL, lupeol, chicoric acid and lupeol+CA decreased the LPS-induced ROS and NO production without cell toxicity in RAW264.7 cells. CGE increased the immune cell populations of $CD4^+T$, $CD8^+T$ and macrophages in various immune organ of mice. In tumor bearing mice, CGAL, lupeol, chicoric acid and lupeol+CA suppressed tumor volume and weight. In cell cycle analysis, they decreased the percentages of S phase. In addition, CGAL, lupeol, chicoric acid and lupeol+CA immune cell populations of $CD4^+T$, $CD8^+Tcell$, NK cell and macrophage in tumor as well as NK cell activity. Conclusion : CGAL and its compounds may enhance immune responses and suppress tumor growth, and may be capable of developing health functional foods.

• 동의생리병리학회지
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• 제23권5호
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• pp.1106-1115
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• 2009

Mori Ramulus has multiple applications in Korean traditional medicine prescription because it has antioxidant and anti-inflammatory effects by reducing macrophage activities. Yet, no studies on the anti-arthritic activity of EMR (extract of Mori Ramulus) have been reported in vitro and in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Because collagen-induced arthritis (CIA) is similar to RA in pathological symptoms and immune reactions, there have been several reports concerning RA using CIA mouse model. Here, we investigated the effects of Mori Ramulus on RA using CIA mice. The importance of CD4+ Th1 cells in RA progress was previously indicated and studies further showed that Th17 cells play a prime role in severity of disease. Accordingly, the present study was focused on CIA associated with CD4+ Th1 cells and Th1 7 cells. DBA/1OlaHsd mice were immunized with bovine type II collagen (CII). After a second collagen immunization, mice were treated with EMR once a day for 4 weeks. The severity of arthritis within the paw joints was evaluated by histological assessment of cartilage destruction and pannus formation. Immune cells in peripheral blood mononuclear cells (PBMC), draining lymph node (DLN) and paw joints, cytokine production and gene expression were assessed from CIA mouse using ELISA, FACS and real-time PCR analysis. Administration of EMR significantly suppressed the progression of CIA and inhibited the production of TNF-$\alpha$, IL-6 and IL-17 in the serum. The erosion of cartilage was dramatically reduced in mouse knees after treatment with EMR. In conclusion, our results demonstrate that EMR significantly suppressed the progression of CIA and that this action was mediated by the decreased production of TNF-$\alpha$, IL-6, IL-17 and collagen II-specific antibody in the serum. EMR suppressed Th17 cells and reduced level of IL-6 via B cell suppression, and thus, the levels of autoantibodies produced from B cells were decreased. Furthermore, EMR suppressed NKT cells which directly stimulate B cells and develop imbalance of Th1/Th2 cell. Oral administration of EMR (100 mg/kg or 200 mg/kg) significantly suppressed the progression of CIA, which is comparable to that of methotrexate (MTX, 0.3 mg/kg) used as a positive control. We are currently studying the mechanism underlying the therapeutic role for EMR in CIA mice.

• 대한약침학회지
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• 제15권2호
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• pp.7-10
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• 2012

Objectives: The root of Lithospermum erythrorhizon Sieb. et Zucc. (Lithospermi Radix, LR) is a kind of heat clearing and blood cooling medicinal herbs. It can clear away heat and cool the blood, reduce toxins and disperse maculae. LR has long been used as efficacious therapy for inflammation, burns, frostbite and skin diseases such as eczema and psoriasis. Methods: In the present study, we investigate anti-allergic and anti-inflammatory effects of LR by using the 1-fluoro-2, 4- dinitrofluorobenzene (DNFB)-induced contact dermatitis mouse model. Results: Topical application of 10 mg/mL of LR effectively inhibited skin lesions induced by repeated paintings with DNFB. Topical application of LR also inhibited hyperplasia, edema, spongiosis and infiltrations of mononuclear cells. In addition, production levels of total immunoglobulin and IgG1 in serum were decreased by using LR in vivo. Conclusions: These data suggest that LR acts as an antiinflammatory agent, improving skin lesions in CD mice.

• IMMUNE NETWORK
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• 제10권1호
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• pp.26-34
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• 2010

Background: Hypersensitivity pneumonitis (HP) is an interstitial lung disease that develops following repeated exposure to inhaled particulate antigens. The family of $Fc{\gamma}$ receptors ($Fc{\gamma}Rs$) has emerged as central regulators for modulating both pro-and anti-inflammatory responses. However, the role of $Fc{\gamma}Rs$ in the development of HP has not been investigated yet. Methods: To explore the functional roles of $Fc{\gamma}Rs$ in HP, $Fc{\gamma}R^{-/-}$ and B6 mice were challenged with Saccharopolyspora rectivirgula (SR) antigen intranasally, and compared these mice in terms of the histological change, infiltrated immune cells in BALF and in vitro immune responses. Results: $Fc{\gamma}R^{-/-}$ mice exhibited attenuation of HP in terms of histological alterations, and reduced numbers of neutrophils and macrophages in and the increased CD4 : CD8 ratio of bronchoalveolar lavage fluid. The lungs of $Fc{\gamma}R^{-/-}$ mice showed high production of Th2 cytokine such as IL-4 and slightly low production of Th1 cytokine, INF-${\gamma}$ compared to those of B6 mice. However, SR-specific adaptive immune responses of $Fc{\gamma}R^{-/-}$ mice were similar to those of B6 mice. Conclusion: These results demonstrate that activating $Fc{\gamma}$ receptors play an important role in activating neutrophils and macrophages in pulmonary inflammation and inducing Th1 differentiation by regulating cytokine expression in SR-induced HP.

• Animal cells and systems
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• 제12권1호
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• pp.15-21
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• 2008

Nitric oxide(NO) has been known to play important roles in numerous physiologic processes including neurotransmission, vasorelaxation, and cellular apoptosis. Using a mouse cDNA gene chip, we examined expression patterns and time course of NO-dependent genes in mouse macrophage RAW264.7 cells. Genes shown to be upregulated more than two fold or at least at two serial time points were further selected and validated by RT-PCR. Finally, 81 selected genes were classified by function as signaling, apoptosis, inflammation, transcription, translation, ionic homeostasis and metabolism. Among those, genes related with signaling, apoptosis and inflammation, such as guanylate cyclase 1, soluble, alpha3(Gucy1a3); protein kinase C, alpha($Pkc{\alpha}$); lymphocyte protein tyrosine kinase(Lck); BCL2/adenovirus E1B 19 kDa-interacting protein(Bnip3); apoptotic protease activating factor 1(Apaf1); X-linked inhibitor of apoptosis(Xiap); cyclin G1(Ccng1); chemokine(C-C motif) ligand 4(Ccl4); B cell translocation gene 2, anti-proliferative(Btg2); lysozyme 2(Lyz2); secreted phosphoprotein 1(Spp1); heme oxygenase(decycling) 1(Hmox1); CD14 antigen(Cd14); and granulin(Grn) may play important roles in NO-dependent responses in murine macrophages.

• 대한한의학회지
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• 제39권1호
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• pp.1-12
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• 2018

Objectives: Astragalus membranaceus has been reported to inhibit immune responses, but its effect on hair loss is not clear. In this study, the effect of A. membranaceus extract (AM) on hair regrowth in C57BL/6 mice with natural hair loss in the telogen phase was investigated. Methods: Mice with natural hair loss were topically treated with 1% AM on the dorsal skin for 2 weeks. Dorsal skin samples were stained with hematoxylin and eosin and probed with an anti-mouse CD8a IgG. The mRNA expression levels of tumor necrosis factor $(TNF)-{\alpha}$, interferon $(IFN)-{\gamma}$ and interleukin (IL)-4 were measured by reverse transcription polymerase chain reaction and quantitative real-time polymerase chain reaction. Results: AM treatment induced hair regrowth in hair loss mice, while control mice suffered continued hair loss. Tapering hair shafts and broken hair follicles were decreased as well as CD8+ T lymphocyte infiltration. In addition, the expressions of $TNF-{\alpha}$, $IFN-{\gamma}$ and IL-4 were reduced by AM treatment. Also, AM treatment significantly increased the KGF expressions in Hs68 fibroblast cells. Conclusion: These results suggest that topical application of A. membranaceus may be an alternative therapy for hair loss.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.256-261
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• 2004

In order to detect chitooligosaccharides (COS) in soybean paste, tandem immunoaffinity chromatography and enzyme-linked immunosorbent assay (ELISA) were developed. Polyclonal anti-chitooligosaccharides mixture (CaSM) antibody specific to COSM was attached to Sepharose gel for initial sample cleanup and concentration of COS in soybean paste. COS was eluted and quantified by competitive direct ELISA (cdELISA). Average ELISA recoveries from the column using binding buffer spiked with COSM at levels of 0.5, 2.0, 5.0, and $10.0\mu$g/ml were 79.8, 72.0, 77.7, and 60.6%, respectively, with a mean recovery of 72.5%. Mean inter-well and inter-assay coefficients of variation (CV) were 7.7% and 10.3%, respectively. Average recoveries from soybean paste spiked with COSM at levels of 2, 6, 20, and $60\mu$g/g were 115, 91.7, 91, and 73.3%, respectively, with a mean recovery of 92.8%. Mean inter-well and inter-assay CV were 12.9% and 16%, respectively. The COS was detected from 24 out of 25 homemade Korean soybean paste samples at an average of $14.0\mu$g/g (n, 25; range, $0-51.2 \mu$g/g) and from 13 out of 14 commercially made soybean paste samples at an average of $4.1\mu$g/g(n, 14; range, $0-18.4\mu$g/g). The tandem immunoaffinity chromatography-cdELISA that was developed in this study showed that the level of COS eluted from homemade soybean paste was higher than that of the commercially made ones. In addition, the level of COS eluted from commercially available soybean paste in Korea was higher than that of the ones in Japan.

• 융합정보논문지
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• 제9권8호
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• pp.274-281
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• 2019

본 연구는 하수오에서 분리한 PM-E와 PM-70M를 이용하여 GATA-3의존성 Th2세포 조절과 IgE 억제 효과를 실험하였다. 하수오를 물질분리하여 GATA-3 전사인자 활성을 억제하는 하수오 물질분획층이 흡착크로마토그래피를 통하여 70% 메탄올층에 포함되어 있다고 분석하였다. 그 결과 PM-70 %MFL 분획이 Th2 사이토카인을 조절하는 GATA-3를 억제하여 항알레르기 작용 효과가 있다고 생각된다. 하수오를 흡착크로마토그래피로 분리하여 분획들을 anti-CD40/rmIL-4와 동시 배양하여 B세포에서 IgE 분비를 억제하는지를 알아본 결과, PM-30M층, PM-70A, 그리고 PM-30A층의 IL-6와 TNF-${\alpha}$ mRNA, IgE 분비량은 대조군과 차이가 없었다. 그러나 IL-6와 TNF-${\alpha}$ mRNA 유전자 발현은 각각 $0.69{\pm}0.058$(p<0.001)과 $0.72{\pm}0.58$ (p<0.05)로 대조군에 비하여 30% 이상 유의하게 억제하였다. IgE 분비량은 $94.6{\pm}16.0$으로 대조군에 비교하였을 때 45.6% 이상 유의하게 감소를 나타내었다 (p<0.01). 본 연구를 통해 하수오의 분획중 PM-70 %MFL층에 B세포의 분화 및 활성을 억제하는 성분이 포함된 것으로 사료된다.

• Tuberculosis and Respiratory Diseases
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• 제45권5호
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• pp.1047-1057
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• 1998

연구배경: 유육종증은 전신을 침범하는 만성 육아종성 질환으로 단핵구성 폐포염이 선행되어 육아종형성을 유도하며, 여러 종류의 cytokine들이 육아종성 염증과 그에 따른 섬유화의명태생리가전에 관여한다고 알려져 있다. 특히 최근에는 IL-6와 IL-8이 유육종증에서 광범위하게 염증반응을 진행시키는 것으로 보고되고 있고, 반면에 IL-10은 단핵구/대식세포와 T림프구의 기능을 억제하여 IL-1, IL-6, TNF-$\alpha$, IFN-$\gamma$ 등의 분비를 감소시키므로써 육아종성 염증반응을 억제한다고 보고되고 있다. 본 연구에서는 유육종증환자들의 폐포세척액내에서 IL-6, IL-8, IL-10의 농도를 측정하고 유육종증의 활성화를 나타내는 임상적 지표들과 비교해 봄으로써 이러한 lnterleukin들이 유육종증의 병태생리기전에 관여하는지 알아보고자 하였다. 대상 및 방법: 폐조직검사를 통해 조직학적으로 확진된 유육종증환자 14명과 건강한 정상대조군 6명을 대상으로 기관지폐포세척술을 시행하였다. 유육종증환자들은 중요 장기(심장, 중추신경, 후방 포도막등)의 침범여부, 고칼숨혈증, 심각한 뼈기능감소 및 질환의 진행여부에 따라 활동성(n=7)과 비활동성(n=7)으로 분류하였고 회수한 폐포세척액을 동일한 조건에서 10배로 농축한 후 ELISA방법을 이용하여 IL-6, IL-8, IL-10을 측정하여 이 Interleukin들의 상호관계와 유육종증의 활성도를 비교하였다. 결 과: 기관지폐포세척액내 IL-6농도는 특히 활동성 유육종증환자군에서 비활동성 유육종증환자군 및 정상대조군보다 유의하게 높았다(각각 p<0.05). IL-8 농도는 활동성 유육종증환자군에서 비활동성 유육종증환자군 및 정상대조군보다 높은 경향을 보였으나 통계적 유의성은 없었고 IL-10농도는 특히 활동성 유육종증환자군에서 정상대조군 및 비활동성 유육종증환자군보다 낮은 경향을 보였으나 통계적 유의성은 없었다. 활동성 유육종증환자군의 기관지폐포세척액내 IL-6농도는 기관지폐포세척액내 IL-8 농도 및 CD4/CD8 비(比)와 서로 의미 있는 상관관계를 보였다(p<0.05). 결 론: 활동성 유육종증환자군의 기관지폐포세척액내에서 IL-6농도가 비활동성 유육종증환자군과 정상대조군에서 보다 유의하게 높았고 이러한 IL-6는 기관지폐포세척액내 CD4/CD8 비(比) 및 IL-8농도와도 상관관계를 보여, 주로 IL-6이 유육종증의 진행과정과 활성화에 종요한 역할을 하고 있는 것으로 생각된다. 또한 세포성 면역반응을 억제할 수 있는 cytokine으로 생각되는 IL-10의 농도가 유육종증환자의 폐내에서 증가되지 않은 것도 이 질환의 발병이나 진행에 기여할 것으로 추정되며 앞으로 유육종증의 치료에 IL-6의 단일클론성 항체나 IL-10이 이용될 수도 있을 것으로 생각된다.