• 생명과학회지
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• 제32권7호
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• pp.491-500
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• 2022

HK표고버섯균사체(HK shiitake mushroom mycelium, HKSMM)는 간 건강 개별 인정 건강기능식품이다. LPS로 활성화된 RAW 264.7 세포에서 HKSMM50 (HKSMM의 50% ethanol 수용액 추출물)의 항염증효과를 연구하였다. AHCC는 positive control로 사용하였다. LPS로 활성화된 RAW 264.7 세포에 HKSMM50 및 AHCC를 처리(0, 20, 100, 500 ㎍/ml)하고 24시간 배양하여 배양물의 염증 관련 인자는 ELISA kits로, 세포에 함유된 iNOS와 COX-2 protein 발현은 Western blotting으로 측정하였다. HKSMM50는 LPS 처리에 비해 농도 의존적으로 NF-κB 함량을 낮추었고, iNOS와 COX-2 protein 발현을 억제하여 NO와 PGE₂ 함량을 낮추었다. 더불어 HKSMM50는 LPS 처리에 비해 IL-1β, TNF-α, IL-4 및 IL-6의 함량을 낮추었으나 SOD와 CAT의 활성은 증가시켰다. AHCC도 HKSSM50 처리와 비슷한 효과를 나타내었다. 이 결과는 HKSMM50이 LPS로 활성화된 RAW 264.7 세포에서 NF-κB 신호전달을 억제하여 항염증효과를 나타내었으며, HKSMM은 면역기능증진에 도움을 줄 수 있는 건강기능식품원료로 사용할 수 있을 것이다.

• BMB Reports
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• 제37권4호
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• pp.507-514
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• 2004

Gamma irradiation ($\gamma$-IR) is reported to have diverse effects on immune cell apoptosis, survival and differentiation. In the present study, the immunomodulatory effect of a low dose $\gamma$-IR (5~10 Gy) was investigated, focusing on the role of NF-${\kappa}B$ in the induction of the B cell differentiation molecule, CD23/FceRII. In the human B cell line Ramos, $\gamma$-IR not only induced CD23 expression, but also augmented the IL-4-induced surface CD23 levels. While $\gamma$-IR did not cause STAT6 activation in these cells, it did induce both DNA binding and the transcriptional activity of NF-${\kappa}B$ in the $I{\kappa}B$ degradation-dependent manner. It was subsequently found that different NF-${\kappa}B$ regulating signals modulated the $\gamma$-IR-or IL-4-induced CD23 expression. Inhibitors of NF-${\kappa}B$ activation, such as PDTC and MG132, suppressed the $\gamma$-IR-mediated CD23 expression. In contrast, Ras, which potentiates $\gamma$-IR-induced NF-${\kappa}B$ activity in these cells, further augmented the $\gamma$-IR- or IL-4-induced CD23 levels, The induction of NF-${\kappa}B$ activation and the subsequent up-regulation of CD23 expression by $\gamma$-IR were also observed in monocytic cells. These results suggest that $\gamma$-IR, at specific dosages, can modulate immune cell differentiation through the activation of NF-${\kappa}B$, and this potentially affects the immune inflammatory response that is mediated by cytokines.

• The Korean Journal of Physiology and Pharmacology
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• 제25권4호
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• pp.307-319
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• 2021

Dysregulation of the Wnt pathway causes various diseases including cancer, Parkinson's disease, Alzheimer's disease, schizophrenia, osteoporosis, obesity and chronic kidney diseases. The modulation of dysregulated Wnt pathway is absolutely necessary. In the present study, we evaluated the anti-inflammatory effect and the mechanism of action of Wnt-C59, a Wnt signaling inhibitor, in lipopolysaccharide (LPS)-stimulated epithelial cells and macrophage cells. Wnt-C59 showed a dose-dependent anti-inflammatory effect by suppressing the expression of proinflammatory cytokines including IL6, CCL2, IL1A, IL1B, and TNF in LPS-stimulated cells. The dysregulation of the Wnt/β-catenin pathway in LPS stimulated cells was suppressed by WntC59 treatment. The level of β-catenin, the executor protein of Wnt/β-catenin pathway, was elevated by LPS and suppressed by Wnt-C59. Overexpression of β-catenin rescued the suppressive effect of Wnt-C59 on proinflammatory cytokine expression and nuclear factor-kappa B (NF-κB) activity. We found that the interaction between β-catenin and NF-κB, measured by co-immunoprecipitation assay, was elevated by LPS and suppressed by Wnt-C59 treatment. Both NF-κB activity for its target DNA binding and the reporter activity of NF-κB-responsive promoter showed identical patterns with the interaction between β-catenin and NF-κB. Altogether, our findings suggest that the anti-inflammatory effect of Wnt-C59 is mediated by the reduction of the cellular level of β-catenin and the interaction between β-catenin and NF-κB, which results in the suppressions of the NF-κB activity and proinflammatory cytokine expression.

• 생약학회지
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• 제39권2호
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• pp.95-103
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• 2008

In the present study, we investigated the anti-inflammatory effects by kaempferol-3-O-${\beta}$-D-sophoroside (KS) isolated from Sophora japonica (Leguminosae) on the lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin ($PGE_2$) production by RAW 264.7 cell line compared with kaempferol. KS significantly inhibited the LPS-induced NO and $PGE_2$ production. Consistent with these observations, KS reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the protein and mRNA levels in a concentration-dependent manner. In addition, the release and the mRNA expression levels of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and interleukin-6 (IL-6) were also reduced by KS. Moreover, KS attenuated the LPS-induced activation of nuclear factor-kappa B ($NF{-\kappa}B$), a transcription factor necessary for pro-inflammatory mediators, iNOS, COX-2, $TNF-{\alpha}$ and IL-6 expression. These results suggest that the down regulation of iNOS, COX-2, $TNF-{\alpha}$, and IL-6 expression by KS are achieved by the downregulation of $NF{-\kappa}B$ activity, and that is also responsible for its anti-inflammatory effects.

• Natural Product Sciences
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• 제14권3호
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• pp.206-213
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• 2008

This study was designed to investigate the anti-inflammatory effects of mangiferin isolated from the rhizome of Anemarrhena asphodeloides, a natural polyphenol, on lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Mangiferin dose-dependently inhibited LPS-induced nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ productions in RAW 264.7 macrophages and peritoneal macrophages isolated from C57BL/6 mice. Consistent with these data, mangiferin suppressed the LPS-induced expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the protein and mRNA levels in a concentration-dependent manner, as determined by Western blotting and RT-PCR, respectively. In addition, the release of tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) and interleukin-6 (IL-6), and the mRNA expression levels of these cytokines were reduced by mangiferin in a dose-dependent manner. Moreover, mangiferin effectively inhibited the transcriptional activation of nuclear factor-kappa B $(NF-{\kappa}B)$. These results suggest that the anti-inflammatory properties of mangiferin are caused by iNOS, COX-2, $TNF-{\alpha}$, and IL-6 down-regulation due to $(NF-{\kappa}B)$ inhibition in RAW 264.7 macrophages.

• 한국자원식물학회지
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• 제33권3호
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• pp.183-193
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• 2020

Recently, as a natural substance has been emphasized interest in research to enhance the immune function. Green lettuce (Lactuca sativa L.) is a popular vegetable used fresh and it contains various phytochemicals and antioxidant compounds, and has been reported to have various physiological activities such as antibacterial, antioxidant, antitumor and anti-mutagenic. However, only a few studies have investigated on the mechanism of action of immune-enhancing activity of lettuce. Therefore, in this study, the immunomodulatory activities and potential mechanism of action of Green lettuce extracts (GLE) were evaluated in the murine macrophage cell line RAW264.7. GLE significantly increased NO levels by RAW264.7 cells, as well as expressions of immunomodulators such as iNOS, COX-2, IL-1β, IL-6, IL-12, TNF-α and MCP-1. Although GLE activated ERK1/2, p38, JNK and NF-κB, GLE-mediated expressions of immunomodulators was dependent on p38, JNK and NF-κB. In addition, TLR4 inhibition blocked GLE-mediated expressions of immunomodulators and activation of p38, JNK and NF-κB. Taken together, these results demonstrated that TLR4-MAPK/NF-κB signalling pathways participated in GLE-induced macrophage activation and GLE could be developed as a potential immunomodulating functional food.

• The Korean Journal of Physiology and Pharmacology
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• 제22권4호
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• pp.369-377
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• 2018

Spinal tuberculosis (ST) is the tuberculosis caused by Mycobacterium tuberculosis (Mtb) infections in spinal curds. Isoliquiritigenin (4,2',4'-trihydroxychalcone, ISL) is an anti-inflammatory flavonoid derived from licorice (Glycyrrhiza uralensis), a Chinese traditional medicine. In this study, we evaluated the potential of ISL in treating ST in New Zealand white rabbit models. In the model, rabbits (n=40) were infected with Mtb strain H37Rv or not in their $6^{th}$ lumbar vertebral bodies. Since the day of infection, rabbits were treated with 20 mg/kg and 100 mg/kg of ISL respectively. After 10 weeks of treatments, the adjacent vertebral bone tissues of rabbits were analyzed through Hematoxylin-Eosin staining. The relative expression of Monocyte chemoattractant protein-1 (MCP-1/CCL2), transcription factor ${\kappa}B$ ($NF-{\kappa}B$) p65 in lymphocytes were verified through reverse transcription quantitative real-time PCR (RT-qPCR), western blotting and enzyme-linked immunosorbent assays (ELISA). The serum level of interleukin (IL)-2, IL-4, IL-10 and interferon ${\gamma}$ ($IFN-{\gamma}$) were evaluated through ELISA. The effects of ISL on the phosphorylation of $I{\kappa}B{\alpha}$, $IKK{\alpha}/{\beta}$ and p65 in $NF-{\kappa}B$ signaling pathways were assessed through western blotting. In the results, ISL has been shown to effectively attenuate the granulation inside adjacent vertebral tissues. The relative level of MCP-1, p65 and IL-4 and IL-10 were retrieved. $NF-{\kappa}B$ signaling was inhibited, in which the phosphorylation of p65, $I{\kappa}B{\alpha}$ and $IKK{\alpha}/{\beta}$ were suppressed whereas the level of $I{\kappa}B{\alpha}$ were elevated. In conclusion, ISL might be an effective drug that inhibited the formation of granulomas through downregulating MCP-1, $NF-{\kappa}B$, IL-4 and IL-10 in treating ST.

• Tuberculosis and Respiratory Diseases
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• 제53권5호
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• pp.519-529
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• 2002

연구배경 : 급성폐손상의 치료로 시도되는 표면활성물질의 효과는 허탈된 폐포를 재환기시키는 작용 외에 표면 활성물질 자제가 가지고 있는 항염증작용이 중요한 기전으로 생각되고 있다. 본 연구에서는 백서의 급성폐손상 모델을 이용하여 기관 내로 표면활성물질을 투여하였을 때, 기관지폐포세척액의 백혈구수와, 염증매개 사이토카인인 IL-$1{\beta}$ 그리고 IL-6의 농도에 변화가 있는지를 살펴보고, surfactant의 항염증작용이 전사인자인 NF-${\kappa}B$의 활성의 억제를 통하여 이루어지는지 여부를 Electrophoretic mobility shift assay (EMSA) 법으로 확인하였다. 방 법 : 대상동물은 300g 내외의 수컷 백서를 이용하였으며, 대상동물을 각각 6 마리씩 세 군으로 나누었다. 대조군은 기관 내로 생리식염수(3ml/kg)를 30분 간격으로 투여하였다. 나머지 두 군은 기관 내로 내독소(5mg/kg)를 투여하여 급성폐손상을 유발하고, 30 분 후에 표변활성 물질 치료군은 surfactant(30mg/kg)을 그리고 비치료군은 생리식염수(3ml/kg)을 각각 기관 내로 투여하였다. 생리식염수나 내독소를 투여한 24 시간 후에 기관지폐포세척술을 시행하였고, 기관지폐포세척액 내의 백혈구수와 IL-$1{\beta}$ 그리고 IL-6의 농도를 측정하였다. 또한 기관지폐포세척액에서 호중구를 분리하고 핵 단백질을 추출하여 NF-${\kappa}B$의 활성을 EMSA법으로 측정하였다. 결 과 : 대조군, 표면활성물질 치료군, 그리고 비치료군의 기관지폐포세척액의 백혈구 수는 각각 $356{\pm}275{\times}10^3/{\mu}1$, $3,221{\pm}1,914{\times}10^3/{\mu}1$, 그리고 $5,561{\pm}1,757{\times}10^3/{\mu}l$으로 비치료군의 백혈구 수가 가장 높았고, 다음으로 표면활성물질 치료군, 비치료군의 순서였다(p<0.05). 기관지폐포세척액의 IL-$1{\beta}$ 농도는 대조군은 0pg/ml, 표면활성물질 치료군은 $360{\pm}234pg/ml$, 그리고 비치료군은 $2,064{\pm}1,082pg/ml$로 대조군에 비해 표면활성물질 치료군이, 그리고 표면활성물질 치료군에 비해 비치료군의 IL-$1{\beta}$농도가 높았다(p<0.05). 기관지폐포세척액의 IL-6 농도는 대조군, 표면활성물질 치료군, 비치료군에서 각각 $49{\pm}62pg/ml$, $1,754{\pm}1,340pg/ml$, 그리고 $3,621{\pm}567pg/ml$으로 대조군에 비해 표면활성물질 치료군이, 그리고 표면활성물질 치료군에 비해 비치료군의 농도가 높았다(p<0.05). 표면활성물질 치료군과 비치료군 사이에서 호중구의 NF-${\kappa}B$ 활성화에는 차이가 없었다. 결 론 : 이상의 연구로 내독소의 기관 내 투여로 유발한 백서의 급성폐손상에서 기관 내로 투여한 표면활성물질은 기관지폐포세척액의 백혈구수와 염증매개 사이토카인인 IL-$1{\kappa}$ 그리고 IL-6의 농도를 감소시켜 폐포 내의 염증을 감소시켰으며, 표면활성 물질의 항염증작용은 NF-${\kappa}B$의 활성의 억제를 통하여 이루어지지는 않는 것으로 판단된다.

• Nutrition Research and Practice
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• 제14권5호
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• pp.453-462
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• 2020

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG), an oriental herbal medicine, has been known to improve liver function, and has both anti-inflammatory and antimicrobial properties. However, little is known about the immune-enhancing effects of PG and its mechanism. In this study, we aimed to investigate whether fermented PG extract (FPGE), which has increased platycodin D content, activates the immune response in a murine macrophage cell line, RAW 264.7. MATERIALS/METHODS: Cell viability was determined by Cell Counting Kit-8 assay and the nitric oxide (NO) levels were measured using Griess reagent. Cytokine messenger RNA levels of were monitored by quantitative reverse transcription polymerase chain reaction. To investigate the molecular mechanisms underlying immunomodulatory actions of FPGE in RAW 264.7 cells, we have conducted luciferase reporter gene assay and western blotting. RESULTS: We found that FPGE treatment induced macrophage cell proliferation in a dose-dependent manner. FPGE also modulated the expression of NO and pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. The activation and phosphorylation levels of nuclear factor kappa B (NF-κB) were increased by FPGE treatment. Moreover, 5-aminoimidazole-4-carboxamide ribonucleotide, an activator of AMP-activated kinase (AMPK), significantly reduced both lipopolysaccharides- and FPGE-induced NF-κB reporter gene activity. CONCLUSIONS: Taken together, our findings suggest that FPGE may be a novel immune-enhancing agent acting via AMPK-NF-κB signaling pathway.

• Nutrition Research and Practice
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• 제14권2호
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• pp.109-116
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• 2020

BACKGROUND/OBJECTIVES: Excessive intake of simple sugars induces obesity and increases the risk of inflammation. Thus, interest in alternative sweeteners as a sugar substitute is increasing. The purpose of this study was to determine the effect of saccharin on inflammation in 3T3-L1 adipocytes. MATERIALS/METHODS: 3T3-L1 preadipocytes were differentiated into adipocytes. The adipocytes were treated with saccharin (0, 50, 100, and 200 ㎍/mL) for 24 h. Inflammation was induced by exposure of treated adipocytes to lipopolysaccharide (LPS) for 18 h and cell proliferation was measured. The concentration of nitric oxide (NO) was measured by using Griess reagent. Protein expressions of nuclear factor kappa B (NF-κB) and inhibitor κB (IκB) were determined by western blot analysis. The mRNA expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin 1β (IL-1β), interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor-α (TNF-α) were determined by real-time PCR. RESULTS: Compared with the control group, the amount of NO and the mRNA expression of iNOS in the LPS-treated group were increased by about 17.6% and 46.9%, respectively, (P < 0.05), and those parameter levels were significantly decreased by saccharin treatment (P < 0.05). Protein expression of NF-κB was decreased and that of IκB was increased by saccharin treatment (P < 0.05). Saccharin decreased the mRNA expression of COX-2 and the inflammation cytokines (IL-1β, IL-6, MCP-1, and TNF-α) (P < 0.05). CONCLUSIONS: The results of this study suggest that saccharin can inhibit LPS-induced inflammatory responses in 3T3-L1 adipocytes via the NF-κB pathway.