• The Korean Journal of Physiology and Pharmacology
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• 제6권6호
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• pp.319-325
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• 2002

The induction of interleukin-6 (IL-6) using combined proinflammatory agents $(LPS/IFN-{\gamma}\;or\;TNF-{\alpha}/IFN-{\gamma})$ was studied in relation to p38 mitogen-activated protein kinase (MAPK) and $NF-{\kappa}B$ transcriptional factor in primary neonatal cardiomyocytes. When added to cultures of cardiomyocytes, the combined agents $(LPS/IFN-[\gamma}\;or\;TNF-{\alpha}/IFN-{\gamma})$ had stimulatory effect on the production of IL-6 and the elevation was significantly reduced by SB203580, a specific p38 MAPK inhibitor. SB203580 inhibited protein production and gene expression of IL-6 in a concentration-dependent manner. In this study, $IFN-{\gamma}$ enhancement of $TNF-{\alpha}-induced\;NF-{\kappa}B$ binding affinity as well as p38 MAP kinase activation was observed. However, a specific inhibitor of p38 MAPK, SB203580, had no effect on $TNF-{\alpha}/IFN-{\gamma}\;or\;LPS/IFN-{\gamma}-induced\;NF-{\kappa}B$ activation. This study strongly suggests that these pathways about $TNF-{\alpha}/IFN-{\gamma}$ or $LPS/IFN-{\gamma}-activated$ IL-6 release can be primarily dissociated in primary neonatal cardiomyocytes.

• 동의생리병리학회지
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• 제23권1호
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• pp.104-112
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• 2009

The purpose of this study was to investigate the anti-inflammatory effects of extract from GuBoEum(GBE) on the peritoneal macrophage. To evaluate anti-inflammatory effects of GBE. I measured cytokines (interleukin-6; IL-6, interleukin-12; IL-12, tumor necrosis factor-$\alpha$; TNF-$\alpha$) and nitric oxide (NO) production in lipopolysacchride (LPS)-induced macrophages. Furthermore, I examined molecular mechanism using western blot and also LPS-induced endotoxin shock. Extract from GBE does not have any cytotoxic effect in the peritoneal macrophages. Extract from GBE reduced LPS-induced IL-6, TNF-$\alpha$, IL-12 and NO production in peritoneal macrophages. GBE inhibited the activation of extracelluar signal-regulated kinase (ERK), C-Jun $NH_2$-terminal kinase (JNK) but not of p38, degradation of $I{\kappa}B-{\alpha}$ in the LPS-stimulated peritoneal macrophages. GBE inhibited the production of TNF-$\alpha$, IL-6 and IL-12 in serum after LPS injection. These results suggest that GBE may inhibit the production of TNF-$\alpha$, IL-6, and IL-12 through inhibition of ERK and JNK activation, and that GBE may be beneficial oriental medicine for inflammatory diseases.

• The Korean Journal of Physiology
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• 제30권1호
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• pp.77-83
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• 1996

Platelet-activating factor(PAF) is a phospholipid mediator of pulmonary inflammation, and immunologic reaction. In this study, the role of PAF on tumor necrosis factor$(TNF_{-{\alpha}})$ production by rat alveolar macrophages(AM) was examined. When PAF $(10^{-12}{\sim}10{-16}\;M)$ alone was added to AM culture, $(TNF_{-{\alpha}})$ production was not significantly increased above the resting level. In contrast, the combined addition of PAF $(10^{-6}\;M)$ and muramyl dipeptide(MDP) $(1.0\;{\mu}g\ml)$ to AM cultures markedly enhanced $(TNF_{-{\alpha}})$ production with 8.2 fold increase compared with AM culture in resting state. This potentiative effect was 313% above the sum of the separate effects of PAF and MDP. To characterize MDP effects on $(TNF_{-{\alpha}})$ production, the dose-response of AM cultured with various concentrations of MDP was tested. High level of MDP $(10\;{\mu}g\ml)$ could not significantly enhance the potentiation effect on $(TNF_{-{\alpha}})$ production compared with AM cultures with low level of MDP $(0.1\;{\mu}g\ml)$, i.e. 112.5% vs 107.8%, respectively when $10^{-10}$ M of PAF was simultaneously added to the cell culture. These data support that the potentiation of TNF. g production in AM culture is mediated by PAF rather than MDf It was also evaluated whether the similar result was obtained in silica, respirable toxic particle-treated AM culture. $(TNF_{-{\alpha}})$ production was also significantly enhanced in the PAF $(10^{-6}\;M)$ and silica $(50\;{\mu}g\ml)$-added cell cultures with 4.7 fold above the value of silica alone-stimulated cells. These results indicate that PAF can potentiate $(TNF_{-{\alpha}})$ production by MDP-or silica- stimulated AM and suggest that PAF may play a potent role in lung inflammation and disease associated with microbe and occupational dust exposures.

• Korean Journal of Acupuncture
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• 제32권3호
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• pp.116-123
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• 2015

Objectives : The present study was designed to investigate effects and molecular mechanisms of Atractylodes macrocephala Koidzumi extracts(AMK) on the improvement of adipocyte dysfunction induced by TNF-${\alpha}$ in 3T3-L1 adipocytes. We examined whether AMK could directly influence the inflammation and insulin resistance in 3T3-L1 adipocytes. Methods : Potential roles of AMK in the lipolysis, production of inflammatory adipokines and ROS, expression and phosphorylation of ERK, JNK, and $I{\kappa}B{\alpha}$ protein, and expression of $PPAR{\gamma}$ and C/EBP${\alpha}$ were investigated in this study. Results : Our data demonstrated that TNF-${\alpha}$ significantly increased lipolysis, levels of MCP-1, IL-6, and ROS and phosphorylation of ERK, JNK, and $I{\kappa}B{\alpha}$ protein, while TNF-${\alpha}$ reduced the expression of $PPAR{\gamma}$ and C/EBP${\alpha}$ in adipocytes, suggesting that TNF-${\alpha}$ induced a condition with the occurrence of inflammation and insulin resistance. Those alterations induced by TNF-${\alpha}$ were prevented by the treatment of AMK. AMK down-regulated the phosphorylation of ERK, JNK, and $I{\kappa}B{\alpha}$ protein and up-regulated the expression of $PPAR{\gamma}$ and C/EBP${\alpha}$ on TNF-${\alpha}$-induced inflammation and insulin resistance. Conclusions : Thus, our results indicate that AMK can be used to prevent from the TNF-${\alpha}$-induced adipocyte dysfunction through MAPK, $NF{\kappa}B$ and $PPAR{\gamma}$ pathways.

• 한국식품영양학회지
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• 제27권1호
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• pp.43-49
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• 2014

생체 내 실험에서 발효 인삼꽃 추출물(FM), 발효하지 않은 인삼꽃 추출물(FD)과 대조군으로 생리식염수를 2주간 마우스 체중 kg 당 100, 200 mg/kg B.W.의 농도로 마우스에 경구 투여한 후 LPS에 의해 활성화된 복강 대식세포가 분비하는 염증성 사이토카인 IL-6, TNF-${\alpha}$의 생성량을 측정하였다. 그 결과, IL-6는 발효 LPS로 자극한 경우, 두 가지 농도에서 모두 처리한 군에 비해 높은 증식능을 나타내었고, LPS로 자극한 결과, 특히 발효 인삼꽃 추출물 200 mg/kg B.W. 농도에서 유의적으로 낮은 IL-6 분비량을 보였다. TNF-${\alpha}$의 경우, 100 mg/kg B.W.와 200 mg/kg B.W. 두 농도 모두에서 LPS로 자극하지 않은 경우, 낮은 증식 효과를 보여주었고, 자극한 경우, 인삼꽃 시료를 첨가한 군이 대조군보다 낮은 TNF-${\alpha}$ 분비량을 보였으며, FM에서 FD보다 더 TNF-${\alpha}$를 억제하는 효과가 큰 것을 볼 수 있었다. 이상의 결과에 따르면 FD의 사이토카인 IL-6, TNF-${\alpha}$ 생성 효과는 200 mg/kg B.W. 농도 투여 시 효과적으로 면역 세포와 면역 기관의 주요 기능을 증진시킬 가능성이 있을 것으로 사료된다. 이러한 연구결과를 토대로 앞으로 인삼꽃 발효를 이용한 기능성 사료 개발과 더불어 산업적 측면에서 보다 긍정적인 효과를 얻을 수 있을 것으로 판단된다.

• 한국식품영양과학회지
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• 제34권10호
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• pp.1503-1508
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• 2005

본 연구에서는 대식세포인 RAW 264.7 세포주에 SQ ($0.1{\%}$), AG($0.1{\%}$), Batyl(1 mg/mL), Chimyl(1 mg/mL) 용액 등을 단독 혹은 C. albicans와 함께 첨가하여 배양한 후LDH, NO, TNF-$\alpha$, IL-6 발현에 어떤 영향을 주는 지를 관찰하였다. 모든 실험군에서 실험에 사용한 농도는 LDH, NO생산에 영향을 주지 못하였다. 실험에 사용한 용액을 단독 처리한 실험의 경우 TNF-$\alpha$와 IL-6발현의 경우 6시간째에는 큰 차이를 나타내지는 못하였으나, 24시간째에는 발현에 영향을 주었다(p < 0.05). C. albicans을 함께 처리한 실험의 경우에는 C. albicans단독 처리군에 비해 실험 용액을 함께 처리한 군에서 C. albicans유도 TNF-$\alpha$와 IL-6발현을 감소시키는 효과를 관찰하였다(p < 0.05, p < 0.01). 본 연구를 통해 SQ, AG, Batyl, Chimyl 용액이 C. albicans 유도에 의해 감소된 면역반응을 증가시킬 수 있을 것으로 생각되어진다.

• Parasites, Hosts and Diseases
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• 제49권2호
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• pp.191-194
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• 2011

Sparganosis is a tissue invading helminthiasis infecting intermediate hosts, including humans. Strong immune responses are expected to occur in early phases of infection. Thus, we investigated cytokine expressions in splenic dendritic cells and in sera after experimental infection of mice. In splenic dendritic cells, TNF-${\alpha}$ and IL-$1{\beta}$ expression peaked at week 1 and week 3 post -infection (PI), respectively, and also early phase (week 2 PI) depressed cytokine expression was noticed. Serum IL-$1{\beta}$ concentration increased significantly at week 2 PI and peaked at week 6 PI, and that of TNF-${\alpha}$ peaked at week 6 PI. These results showed that pro-inflammatory cytokines, TNF-${\alpha}$ and IL-$1{\beta}$, are chronologically regulated in mouse sparganosis.

• Tuberculosis and Respiratory Diseases
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• 제44권3호
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• pp.547-558
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• 1997

연구배경 : 종양괴사인자(tumor necrosis factor ; TNF)는 다양한 생물학적 기능을 가지고 있는바, 그 중 생체외에서 증명된 뚜렷한 항암효과로 말미암아 최근 항암유전자요법의 중요한 대상으로 관심을 모으고 있다. 그러나 유전자 이입의 기술적 문제로 생체외에서 암세포에 유전자 이입을 시행한 후 이를 다시 환자의 생체내로 이식하는 방법이 연구의 주종을 이루고 있다. 그러나 저지들의 과거의 연구를 포함한 여러 연구에서 TNF가 이입된 암세포는 TNF에 대해 내성을 보이는 것으로 증명되었다. 이 획득내성의 기전을 밝히는 것이 종양생물학의 이해를 넓히고 보다 효과적인 항암유전자 요법을 개발하기 위한 매우 중요한 과제로 생각된다. 저자들은 TNF 유전자 이입에 따른 암세포의 TNF에 대한 획득 내성에 새로운 방어단백질의 합성이 관여하는 지를 규명하고자 본 실험을 수행하였다. 방 법 : TNF에 감수성을 보이는 생쥐 섬유육종 세포주인 WEHI164에 TNF-$\alpha$ 유전자를 retroviral vector를 이용하여 이입하고 TNF의 발현을 시도하여 PCR, ELISA, MTT assay로 확인하였고, TNF 유전자가 이입된 세포(WEHI164-TNF)는 TNF에 내성을 보이는지 역시 MTT assay로 검증하였다. WEHI164-TNF세포를 transcription 억제제인 actinomycin D와 translation 억제제인 cycloheximide로 처리한 후 역시 MTT assay로 TNF에 대한 감수성에 변화를 보이는지 확인하였다. 결 과 : 1) TNF-$\alpha$ 유전자 이업 및 발현 확인 PCR을 시행한 결과 TNF 유전자가 이입된 WEHI164-TNF 세포주는 790 base pair 크기의 진한 DNA band를 보인 반면 모세포주는 보이지 않아서 retroviral vector를 이용한 유전자 이입이 DNA 수준에서 이루어졌음을 확인할 수 있었다. 그리고 WEHI164-TNF의 배양상층액에서 TNF양을 ELISA와 MTT assay로 측정한 결과, 생물학적 활성을 지닌 TNF를 $10.9{\pm}1.47ng/24hr/10^6cells$ 생산함을 알 수 있었다. 2) TNF 유전자 이입 전후, 암세포의 TNF에 대한 감수성 비교 TNF 농도 100ng/ml 에서 모세포는 $73{\pm}5%$의 세포독성을 보인 반면 WEHI164-TNF 세포는 $3{\pm}2%$의 세포독성을 보여 통계적으로 유의하게 (p < 0.00) TNF에 대한 내성을 획득함을 알 수 있었다. 3) TNF 유전자 이입 후 획득된 TNF에 대한 내성의 기전 WEHI164-TNF 세포를 actinomycin D로 처리한 경우 TNF 농도 10ng/ml과 100ng/ml에서 각각 $24{\pm}7%$, $44{\pm}6%$의 세포독성을 보여 control의 $6{\pm}2%$, $17{\pm}2%$보다 통계적으로 유의하게(p < 0.01) TNF에 대한 감수성이 부분적으로 회복됨을 관찰할 수 있었다. 그러나 cycloheximide로 처리한 경우에서는 TNF에 대한 감수성에 변화를 관찰할 수 없었다. 결 론 : TNF에 감수성을 보이는 암세포주인 WEHI164에 TNF 유전자를 이입하여 TNF를 발현하게 하였을 때 그 세포 자신은 TNF에 대해 내성을 획득하게되며 이에는 미지의 방어단백질의 합성이 일부 관여할 것으로 판단된다.

• 한국자원식물학회지
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• 제22권6호
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• pp.552-557
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• 2009

Allicin, a garlic componente, is believed to provide protection against various diseases including inflammation. Since interactions of the cell adhesion molecules are known to play important roles in mediating inflammation, inhibiting adhesion protein upregulation is a possible therapeutic target. In this study, we demonstrate that TNF-${\alpha}$- and catalase-induced expression of ICAM-1 on human lung epithelial cells (A549) in a dose-dependent manner and catalase expression and activity were also increased in TNF-${\alpha}$-treated cells. Treatment of the TNF-${\alpha}$-treated cells with catalase inhibitor 3-amino-1,2,4-triazole resulted in a significant decreased the level of ICAM-1. These data suggest that induction of ICAM-1 expression by TNF-${\alpha}$ is associated with catalase. In addition, allicin was found to inhibit the TNF-${\alpha}$ induced expression of ICAM-1 on the A549 cells. This compound also inhibited the production of catalase induced by TNF-${\alpha}$, which suggests that the inhibition of ICAM-1 expression by allicin may be due to the modulated production of catalase.

• 대한수의학회지
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• 제51권3호
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• pp.239-241
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• 2011

In this study, we investigated the kinetics of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6 and high mobility group box 1 (HMGB1) concentrations in a 48-h model of canine endotoxemia by lipopolysaccharide (LPS) injection. Four healthy beagles were slowly administered 1 mg/kg of LPS diluted in normal saline, while two others were administered normal saline as controls. Blood collection was performed at 0 h (baseline), 1 h and 3 h (for TNF-${\alpha}$), 6 h, 12 h, 24 h and 48 h of the experiment, and cytokine levels were determined using the sandwich ELISA method. Early increments of TNF-${\alpha}$ and IL-6 were observed (< 3 h), but HMGB1 levels increased the most at 12 h of the experiment and gradually decreased until 48 h. During the whole experiment, IL-6 and HMGB1 were sustained over 12 h of LPS injection, whereas TNF-${\alpha}$ decreased within 6 h of LPS injection. Taken together, canine HMGB1 levels increase relatively late (< 12 h) and sustained longer than TNF-${\alpha}$ and IL-6 in response to endotoxin. This is the first study to evaluate canine HMGB1 cytokine from endotoxemia in dogs.