• 생약학회지
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• 제37권4호
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• pp.221-228
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• 2006

A lectin was purified from Serrognathus platymelus castanicolor larvae and named as SPL. The purification was carried out by ion-exchange chromatography on DEAE Sephadex A-50 and gel filtration chromatography on Sephadex G-200. The purity of the protein was verified by polyacrylamide gel electrophoresis and the purified lectin agglutinated erythrocytes of rabbit and human A, B, O, AB. SPL was tested it's ability to enhance the expressions of cytokines, $IL-1\alpha$, IL-2, IL-6, $TNF\alpha$ and $IFN\gamma$ by human peripheral blood mononuclear cells (PBMC) obtained from healthy donors. mRNA analyses were performed by RT-PCR at the moment of 1, 4, 8, 24, 48, 72 and 96 h after stimulation of PBMC with purified SPL. The patterns of IL-2 band were slightly expressed from 24 h and the strongest band was appeared at 96 h. The expressions of $IL-1\alpha$ and IL-6 mRNA were strong from 1 to 8 h and those of $TNF\alpha$ were from 48 to 96 h. The mRNA encoding $IFN\gamma$ were not detected. The addition of SPL for macrophage cultures induced production of nitric oxide (NO) by cells in a dose-dependent manner. NO release was partially inhibited by $TNF\alpha$ antibodies. These results suggest that SPL has the ability to enhance cytokine expressions in PBMC and to induce the NO release by TNFa in macrophage cultures from PBMC cultures.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.60-73
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• 2009

Objective : Houttuyniae Herba is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how Houttuyniae Herba affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted from Houttuynia cordata Thunb(HCT) on the mast cell-mediated inflammatory responses. Method : We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of HCT. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. Nuclear and cytoplasmic proteins were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by a luciferase assay. Result : HCT inhibited the PMA + A23187-induced TNF-$\alpha$, IL-6 expression and reduced mRNA of TNF-$\alpha$, IL-6 and IL-8. we observed that HCT suppressed the induction of NF-B activity. In addition, HCT suppressed PMA plus A23187-induced NF-$\kappa$B promoting activity. Conclusion : In this study, we have found that HCT is an inhibitor of NF-$\kappa$B and cytokines on the mast cell-mediated inflammatory responses.

• 대한한의학회지
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• 제22권2호
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• pp.109-119
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• 2001

Objective : There are many reports that acupuncture has thermoregulatory effects on human and animals. To investigate the effect and mechanism of antipyretic action of acupuncture, we observed the body temperature and cytokine expressions in the hypothalamus of rats. Methods : Lipopolysaccharide (LPS, i.p., 2.5mg/kg) was injected to conscious rats (Sprague-Dawley, male, n=4l) to cause hyperthermia and simple needling (stainless steel, 0.25 mm o.d., 5 mm insertion for 10 sec with no manipulation) was performed bilaterally with the measurement of rectal temperature. Next, we sacrificed rats to remove brain and determined the level of mRNA for interleukin-6 (IL-6), $interleukin-1{\beta}{\;}(IL-1{\beta})$, interleukin-2 (IL-2) and $interferon-{\gamma}{\;}(IFN-{\gamma})$ in the hypothalamus by using reverse transcriptase-polymerase chain reaction (RT-PCR). Resul1s : Needling on forepaw (acupoint HT8) and needling on hindpaw (acupoint BL66 and acupoint LR2) significantly inhibited LPS-induced fever of rats (P<0.01, 10 min after treatment respectively), but same treatment on proximal tail (non-acupoint) did not cause any change on fever. The levels of IL-6 and $IL-1{\beta}$ mRNA in the hypothalamus was significantly enhanced by LPS-injection, while the level of IL-6 and $IL-1{\beta}$ mRNA was markedly reduced after treatment on BL66 (P<0.01). Treatment on forepaw reduced it slightly, but not significantly. Equivalent stimulation on proximal tail did not cause any changes. Conclusions : Our results indicate that acupuncture stimulation on various body parts has differential thermoregulatory effects on LPS-induced fever of rats with site-specificity. And, we suggest that its antipyretic action might be accompanied with the suppression of hypothalamic production of pro-inflammatory cytokine of immune system, IL-6 and $IL-1{\beta}$.

• 대한한방소아과학회지
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• 제19권2호
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• pp.13-30
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• 2005

Objective: Danguieumja-gagambang (DGEJGB), a traditional Korean prescription, has been used as therapeutics for allergic diseases such as atopic dermatitis (AD). In this study, we compared with regulatory Effect of Atopic Allergic Reaction by Prescriptions A and by Prescriptions B. Methods : To evaluate and compare the atopic allergic effectiveness of two prescription (A, B) of DGEJGB, the author investigated a possible effect of DGEJGB on mast cell-mediated allergic reaction, cytokine secretion and mRNA expression in vivo and in vitro. Results : Mast cells are a potent source of mediators that regulate the inflammatory response in allergic reaction. In mice orally administered A, B of DGEJGB ( 0.1, 0.1 and 1.0 g/kg) for 1 h, compound 48/80-induced ear swelling was significantly reduced. Significant reduced levels (P < 0.05). of tumor necrosis factor $(TNF)-{\alpha}$ was observed in the human mast cell line (HMC-1) with DGEJGB (A). IL-6 and IL-8 secretion were significantly inhibited by DGEJGB (A, B). In addition, $TNF-{\alpha}$ and IL-8 mRNA expression were reduced by DGEJGB (A) at the dose of 0.01 mg/ml without cell toxicity. Conclusions : These results suggest that DGEJGB (A) contributes to the treatment of atopic allergic reactions rather than DGEJGB (B), and that its action may be due to inhibition of cytokine secretion and mRNA expression HMC-1.

• 한방안이비인후피부과학회지
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• 제24권1호
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• pp.45-63
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• 2011

Objective : Atractyloides Chinensis Rhizome (ACR) is widely used in oriental medicine as a remedy for an inflammation and an allergic disease. However, as yet there is no clear explanation of how ACR affects the production of inflammatory cytokine. This study was to determine the effects of ACR on the mast cell-mediated inflammatory responses. Method : The amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of ACR was measured. The TNF-${\alpha}$ protein levels were analysised by Western blots. The TNF-${\alpha}$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-${\alpha}$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-${\kappa}$B, phospho-I${\kappa}$B and MAPKs were examined by Western blot analysis. The NF-${\kappa}$B promoter activity was examined by a luciferase assay. Results : 1. The expressions of TNF-${\alpha}$ and TNF-${\alpha}$ mRNA were decreased dose-dependently at 0.05-0.2mg/$m\ell$ of ACR and significantly decreased at 0.2mg/$m\ell$. 2. The expressions of IL-6 and IL-6 mRNA were decreased dose-dependently at 0.05-0.2mg/$m\ell$ of ACR and significantly decreased at 0.2mg/$m\ell$. 3. The expressions of IL-8 and IL-8 mRNA were decreased dose-dependently at 0.05-0.2mg/$m\ell$ of ACR and significantly decreased at 0.2mg/$m\ell$ specially. 4. The expressions of Phosphorylated-JNK were decreased, not p38, ERK 5. The expressions of NF-${\kappa}$B were decreased dose-dependently at 0.1-0.2mg/$m\ell$ of ACR. The expressions of Phosphorylated I${\kappa}$B were significantly decreased at 0.2mg/$m\ell$. In addition, ACR suppressed PMA plus A23187-induced NF-${\kappa}$B promoting activity. Conclusion : It is suggested that ACR should suppress through inhibition of NF-${\kappa}$B activity and cytokine production.

• 생명과학회지
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• 제23권4호
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• pp.569-577
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• 2013

간장은 한국의 전통 발효식품으로써 최근 항암 효과, 항균작용, 항산화능, 항혈전 효과 등의 효능이 보고되고 있다. 한국의 간장은 콩과 밀을 주요 원료로 사용한다. 콩의 세포벽으로부터 유래되는 다당류는 효소적 가수분해에 저항성이 있으므로, 발효가 끝난 후에도 간장에 잔존하게 되며 이것을 간장 유래 다당류(Kanjang polysaccharides, KPS)라 부른다. 본 연구에서는 산분해법과 양조법으로 제조된 시판 간장인 A~T의 20종의 투석물로부터 다당류를 제조하고, 비만세포에서 염증성 cytokine의 방출과 mRNA의 발현에 대한 KPS의 효과를 실험함으로써 항염증 효과를 조사하였다. RBL-2H3 세포에서 KPS의 처리는 histamine과 ${\beta}$-hexosaminidase의 방출을 현저히 억제시켰다. 자극된 HMC-1 세포에서 KPS의 처리는 염증성 cytokine인 IL-6, IL-8, TNF-${\alpha}$의 방출 및 mRNA의 발현을 감소시켰다. 특히, 양조간장으로부터 유래된 KPS는 산분해 간장보다 비만세포에서 우수한 항염증 효과를 나타내었다. 따라서, KPS는 알레르기성 염증반응을 억제시키는데 효과적일 것이라 사료된다.

• 생명과학회지
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• 제32권3호
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• pp.222-231
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• 2022

본 연구는 긴병꽃풀 ethyl acetate 분획물의 미백 및 항염증 활성 검증을 통해 화장품 소재로서의 활용 가능성을 확인하고자 하였다. 전자공여능과 ABTS⁺ radical 소거능 측정 결과 최고 농도인 1,000 ㎍/ml에서 각각 89.6%, 88.7%의 활성을 확인할 수 있었다. Tyrosinase 저해활성 측정 결과 최고 농도인 1,000 ㎍/ml에서 ethyl acetate 분획물은 22.3%의 억제활성을 나타내었다. 세포 생존율 측정 결과, 멜라노마 세포와 대식세포에 대해 긴병꽃풀 ethyl acetate 분획물을 처리한 모든 구간에서 80% 이상의 생존율을 보였다. 긴병꽃풀 ethyl acetate 분획물의 단백질 및 mRNA 발현량을 측정하기 위한 western blot과 RT-PCR의 농도구간은 25, 50, 100 ㎍/ml으로 설정하였다. 그 결과 분획물의 농도가 증가함에 따라 발현양이 감소됨을 확인하였으며 미백관련 인자 MITF와 TRP-2의 단백질 발현 억제율이 대조군인 kojic acid에 비해 우수하였고, tyrosinase에 대해서는 100 ㎍/ml에서 29.1%의 가장 낮은 발현량을 보여 매우 뛰어난 mRNA 발현 억제를 확인할 수 있었다. Pro-inflammatory cytokine인 IL-1β, IL-6 및 TNF-α의 단백질 및 mRNA 발현량에 미치는 영향을 확인한 결과, IL-6와 TNF-α의 인자가 같은 농도의 대조군인 Vit. C에 비해 높은 단백질 및 mRNA 억제율을 나타내었다. 이러한 실험결과를 기반으로 긴병꽃풀 ethyl acetate 분획물이 기능성 소재로서의 활용 가능성을 확인함으로써 화장품에 천연 소재로서 응용할 수 있을 것이라 판단되었다.

• Tuberculosis and Respiratory Diseases
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• 제44권3호
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• pp.601-610
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• 1997

연구배경 : 그람음성 간균의 내독소에 의한 sepsis syndrome에서 단핵 식세포는 내독소에 의해 자극받아 여러 종류의 cytokine을 분비함으로써 개체를 방어하는데 있어 매우 중요한 역할을 한다. 그러나 불행히도 cytokine들은 개체를 방어하는 작용 뿐 아니라 TNF등의 경우처럼 심각한 조직손상을 가져오는 효과도 있다. 그러므로 생체 내에서 내독소에 반응하여 cytokine의 분비를 조절하는 기능은 매우 중요하다. 이전의 연구에서 미리 내독소에 노출된 단색 식세포는 내독소로 재자극 시 cytokine 생성능의 저하가 관찰되었는데 이러한 현상을 '내독소 내성'이라고 하며 cytokine 분비조절에 중요한 역할을 하리라 생각되나 그 기전 등에 대해서는 연구가 부족한 상태이다. 방 법 : 생체 외에서 내독소에 대한 내성획득의 조건을 확립하고자 정상인의 말초혈액 단핵구를 10ng/ml의 저농도 내독소로 24시간 전처치한 후 2회 세척하고 다시 1 ng/ml의 내독소로 각각 4시간, 6시간, 24시간 자극하여 TNF-$\alpha$와 IL-8의 단백량을 측정하고 총 RNA를 분비하였다. 내독소 내성 획득 기전을 밝히고자 내독소 전처치 시에 자가혈청, PMA, antiCD14 Ab, Indomethacin, $PGF_2$를 각각 첨가하여 내성획득에 영향을 주는지 알아보았다. TNF-$\alpha$와 IL-8의 단백량은 ELISA를 이용하여 측정하였고 분리한 RNA를 이용하여 TNF-$\alpha$와 IL-8에 대한 Northern blot analysis를 시행하였다. 결 과 : 말초혈액을 저농도 내독소로 전처치하면 TNF-$\alpha$ 단백 생성 및 mRNA 발현을 억제하였으나 IL-8에 대해서는 이러한 현상을 관찰할 수 없었다. 전처치 시에 antiCD14 Ab를 내독소와 같이 준 경우 억제된 TNF-$\alpha$ 생성이 부분적으로 회복되었다. PMA만으로 전처치 하여도 저농도 내독소 전처치와 유사하게 내독소 내성을 유도할 수 있었다. 결 론 : 내독소에 의한 내성획득에는 CD14가 관여하고 Protein kinase C 경로를 통하며 pretranslational 수준에서 조절되는 것으로 생각된다.

• 대한약침학회지
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• 제7권3호
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• pp.59-71
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• 2004

Objectives : In order to investigate effects and immune improvement of distilled white-ginseng herbal extract, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled white-ginseng Herbal Acupuncture at Wisu($Bl_{21}$) and Chung-wan($CV_{12}$) to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR. the control group and the experiment groups show significant increase. For Cox-2, both experiment groups and the normal group showed significant decrease. For Bcl-2, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups 2. For survival time, all of experiment groups didn't show significant differences. 3. IL-2 productivity using Flow cytometry, experiment group I didn't show any significance, For II-4, all of experiment groups showed slight decrease compared to the control group. 4. For IL-2 productivity using ELISA, experiment group I showed slight decrease compared to the control group, experiment group II didn't show any significance. 5. For expression of cytokine mRNA using RT-PCR, significant increase of IL-2 and IL-4 were witnessed in experiment group I compared to the control group. Significant decrease of IL-10 was shonwn in all of experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled white-ginseng Herbal Acupuncture may be further effects in anti-cancer and immune improvement if increasing concentration.

• Biomolecules & Therapeutics
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• 제18권3호
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• pp.286-293
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• 2010

Cytokine production is a sensitive indicator for monitoring perturbations of the immune system by xenobiotics in animals and humans. In the present study, we evaluated the changes in $IFN{\gamma}$, IL-5 and $TNF{\alpha}$ mRNA expression after atrazine (ATZ), perfluorooctanoic acid (PFOA) or zearalenone (ZEA) exposure in Jurkat cells. The IC50 (concentration for a 50% inhibition of cell proliferation) of PFOA and ZEA after 3 days culture were $226.6\;{\mu}M$ and $52.6\;{\mu}M$, respectively. The effects of ATZ on cytokine expression followed in increasing order of $IFN{\gamma}$>IL-5>$TNF{\alpha}$ at $3\;{\mu}M$ and at the lower concentrations the degree of effects on three cytokines were less clear between the cytokines when compared to control level. PFOA had marked increasing effect in order of $IFN{\gamma}$>$TNF{\alpha}$>IL-5 mRNA expression at IC50, and these patterns were continued at the lower concentrations, IC50/2 and IC50/4. ZEA caused the overexpression of cytokine mRNAs in order of IL-5>$IFN{\gamma}$>$TNF{\alpha}$ at both IC50 and IC50/2, and at IC50/4 the overexpression order was IL-5>$TNF{\alpha}$. On other hand, $IFN{\gamma}$ was less distinct compared to the control. These data indicate that ATZ, PFOA and ZEA caused the overtranscription of $IFN{\gamma}$, IL-5 and $TNF{\alpha}$ mRNA, and the overproduction of these cytokines may eventually lead to immune disorders.