• IMMUNE NETWORK
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• 제5권1호
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• pp.30-35
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• 2005

Background: p38 and extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK) signaling are thought to have critical role in lipopolysaccharide (LPS)-induced immune response but the molecular mechanism underlying the induction of these signaling are not clear. Methods: Specific inhibitors for p38, SB203580, and for ERK, PD98059 were used. Cells were stimulated by LPS with or without specific MAPK inhibitors. Results: LPS activated inducible nitric oxide synthase (iNOS), subsequent NO productions, and pro-inflammatory cytokine gene expressions (TNF-${\alpha}$, IL-$1{\beta}$, IL-6, and IL-12). Treatment of both SB203580 and PD98059 decreased LPS-induced NO productions. Concomitant decreases in the expression of iNOS mRNA and protein were detected. SB203580 and PD98059 decreased LPS-induced gene expression of IL-$1{\beta}$ and IL-6. SB203580 increased LPS-induced expression of TNF-${\alpha}$ and IL-12, and reactive oxygen species production, but PD98059 had no effect. Conclusion: These results indicate that both p38 and ERK pathways are involved in LPS-stimulated NO synthesis, and expression of IL-$1{\beta}$ and IL-6. p38 signaling pathways are involved in LPS-induced TNF-${\alpha}$ and IL-12, and reactive oxygen species plays an important role in these signaling in macrophage.

• 동의생리병리학회지
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• 제26권2호
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• pp.175-180
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• 2012

The purpose of this study is to verify the anti-inflammatory effectiveness of Cheongyeoungkeumyeum(CYK) including seven herbal medicines of that major effectiveness is to clear heat, to relieve fire toxicity and to clear damp-heat. To evaluate anti-inflammatory effectiveness of CYK, we measured the production of reactive oxygen species(ROS), nitric oxide(NO) cyclooxygenase-2(COX-2) and in TNF-${\alpha}$ LPS-activated Raw 264.7 cells. Cell viability was determined by MTT assay. The concentrations of ROS and relative level of NO were measured with DPPH assay and Griess reagent, respectively. COX-2 and TNF-${\alpha}$ were detected by enzyme immuno assay(EIA) and enzyme-linked immunosorbent assay(ELISA). As a result, we found that CYK suppressed LPS-induced ROS and NO production in a dose-dependent manner. Also CYK significantly inhibited LPS-induced COX-2 activity and the release of TNF-${\alpha}$. These results indicate that the CYK may have an anti-inflammatory agent for the treatment of various inflammatory disease.

• 한국응용과학기술학회지
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• 제36권3호
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• pp.966-974
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• 2019

Arthrospira platensis has been reported to contain a variety of substances such as phycocyanin, ${\beta}$-carotene, vitamin E and other carotenoids. In this study, zebrafish were treated with indoor cultivation spirulina ethanol extracts(ICAE) to determine toxicity(coagulation rate, hatching rate, heart rate). We used the DCFH-DA staining method to detect the effect of reactive oxygen species(ROS) generation on lipopolysaccharide(LPS)-induced zebrafish embryos ROS various concentrations(0.01, 0.05, 0.1, 0.5mg/ml) of ICAE. Cell toxicity was measured by WST-1 assay on RAW 264.7 macrophage cell line. Also, measured the inhibitory effect of nitric oxide(NO) and prostaglandin $E_2$ ($PGE_2$) production in RAW264.7 macrophages induced by LPS at various concentrations of ICAE. The results of embryo coagulation rate, hatching rate, heart rate of zebrafish at various(0.01, 0.05, 0.1mg/ml) of ICAE was no toxicity. The ICAE treated group had an inhibitory effect on NO and $PGE_2$ production compared and decreased with concentration. The results of this study ethanol extract of Arthrospira platensis has an anti-inflammatory effect and suggest that is worthy of use cosmetics for skin protection.

• BMB Reports
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• 제42권9호
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• pp.574-579
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• 2009

The regulatory effects of honokiol on the cellular responses of macrophages and monocytes were evaluated. Specifically, we investigated the effects of honokiol with respect to lipopolysaccharide (LPS)-induced cytotoxicity, LPS- or phorbol-12-myristate-13-acetate (PMA)-mediated morphological changes, and relevant events (FITC-dextran-induced phagocytic uptake). Honokiol blocked the LPS-induced cytotoxicity of RAW264.7 cells in a dose-dependent manner. In addition, honokiol appeared to block the production of cytotoxic cytokines such as interleukin (IL)-$1{\beta}$ and tumor necrosis factor (TNF)-$\alpha$, nitric oxide (NO), and reactive oxygen species (ROS). Moreover, honokiol strongly prevented the morphological changes in RAW 264.7 and U937 cells that were induced by LPS and PMA. The surface levels of marker proteins, which are up-regulated under the morphological changes of RAW264.7 and U937 cells, were also diminished. The data presented here strongly suggest that the honokiol modulates various cellular responses managed by macrophages and monocytes.

• 한국약용작물학회지
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• 제16권5호
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• pp.313-319
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• 2008

S. barbata의 열수 추출물 (Sb-HWE)은 대표적인 염증과정인 LPS에 의해 활성화된 대식세포로 부터의 NO생성, LPS 매개에 의한 세포사멸작용, 및 FITC-dextran의 대식세포내 탐식작용을 매우 효과적으로 억제하였다. 그러나 본 추출물은 SNP로 유도된 라디칼 소거능은 매우 미약한 것으로 나타났다. NF-${\kappa}B$-매개에 의한 루시퍼라제 활성, 및 NF-${\kappa}B$ 활성 관련 신호전달 단백질 (Akt 및 $I{\kappa}B{\alpha}$)에 대한 저해작용은 관찰되지 않은 것으로 보아 이들 추출물의 대식세포 면역반응 조절 기전은 기존의 알려진 방법과는 다른 기전에 의해 진행되는 것으로 판단된다.

• 한국식품영양과학회지
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• 제44권8호
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• pp.1241-1247
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• 2015

본 연구의 목적은 LPS로 자극한 RAW264.7 세포를 이용하여 크랜베리에서 분리한 폴리페놀 분획물의 항산화능을 분석하는 것이었다. 크랜베리 분말의 에틸 아세테이트 분획(EF)과 메탄올 분획(MF)은 C18-Sep-Pak 카트리지를 사용하여 분리했다. 20시간 동안 LPS로 자극한 RAW264.7 세포는 활성산소종(ROS)과 DNA 손상이 유의하게 증가하였으며, EF, MF 및 TF(EF+MF)로 전처리하고 LPS로 자극한 RAW264.7 세포에서는 ROS와 DNA 손상이 유의적으로 감소했다. 그러나 superoxide dismutase의 활성에는 차이가 없었다.

• Biomolecules & Therapeutics
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• 제23권2호
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• pp.110-118
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• 2015

According to the expansion of lifespan, neuronal disorder based on inflammation has been social problem. Therefore, we isolated shikonin from Lithospermum erythrorhizon and evaluated anti-inflammatory effects of shikonin in lipopolysaccharide (LSP)-stimulated BV2 microglial cells. Shikonin dose-dependently inhibits the expression of the proinflammatory mediators, nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), and tumor necrosis factor-${\kappa}B$ (TNF-${\alpha}$) as well as their main regulatory genes and products such as inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-${\alpha}$ in LPS-stimulated BV2 microglial cells. Additionally, shikonin suppressed the LPS-induced DNA-binding activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) to regulate the key regulatory genes of the proinflammatory mediators, such as iNOS, COX-2, and TNF-${\alpha}$, accompanied with downregulation of reactive oxygen species (ROS) generation. The results indicate that shikonin may downregulate the expression of proinflammatory genes involved in the synthesis of NO, $PGE_2$, and TNF-${\alpha}$ in LPS-treated BV2 microglial cells by suppressing ROS and NF-${\kappa}B$. Taken together, our results revealed that shikonin exerts downregulation of proinflammatory mediators by interference the ROS and NF-${\kappa}B$ signaling pathway.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.98-98
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• 2018

In this study, we evaluated the anti-inflammatory effect of extracts of leaves (ST-L) and branches (ST-B) from Sageretia theezans in LPS-stimulated RAW264.7 cells. ST-L and ST-B significantly inhibited the production of the pro-inflammatory mediators such as NO, iNOS, COX-2, $IL-1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells. ST-L and ST-B blocked LPS-induced degradation of $I{\kappa}B-{\alpha}$ and nuclear accumulation of p65, which resulted to the inhibition of $NF-{\kappa}B$ activation in RAW264.7 cells. ST-L and ST-B also attenuated the phosphorylation of ERK1/2, p38 and JNK in LPS-stimulated RAW264.7 cells. In addition, ST-L and ST-B increased HO-1 expression in RAW264.7 cells, and the inhibition of HO-1 by ZnPP reduced the inhibitory effect of ST-L and ST-B against LPS-induced NO production in RAW264.7 cells. Inhibition of p38 activation and ROS elimination attenuated HO-1 expression by ST-L and ST-B, and ROS elimination inhibited p38 activation induced by ST-L and ST-B. ST-L and ST-B dramatically induced nuclear accumulation of Nrf2, but this was significantly reversed by the inhibition of p38 activation and ROS elimination. Collectively, our results suggest that ST-L and ST-B exerts potential anti-inflammatory activity by suppressing $NF-{\kappa}B$ and MAPK signaling activation, and activating HO-1 expression through the nuclear accumulation of Nrf2 via ROS-dependent p38 activation. These findings suggest that ST-L and ST-B may have great potential for the development of anti-inflammatory drug to treat acute and chronic inflammatory disorders.

• 대한한의학회지
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• 제43권4호
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• pp.52-73
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• 2022

Objectives: The anti-inflammatory and anti-oxidative activities of LCVC (Lonicerae Flos, Citri Pericarpium and Violae Herba Complex) have not been fully elucidated. The purpose of this study was to investigate the mechanisms underlying these effects in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Methods: The evaluation of the anti-oxidative activity of LCVC was completed via DPPH and ABTS radical scavenging capacity, FRAP assay, measurement of polyphenol and flavonoid, assessment of ROS and NO levels in LPS-induced RAW 264.7 cells. The anti-inflammatory activity was defined by measuring the production of biomarkers (PGE2, IL-1B, IL-6 and TNF-𝛼), proteins (ERK, JNK, P38, Nrf2, Keap1, HO-1 and NQO1) and expressions of genes (iNOS, COX-2, IL-1𝛽, IL-6, TNF-𝛼, Nrf2, Keap1, HO-1 and NQO1) in LPS-induced RAW 264.7 cells. Results: LCVC have polyphenol and flavonoid contents. The results of DPPH and ABTS free radical scavenging capacity and FRAP assay showed that the anti-oxidative activity was increased. Production of ROS, NO, IL-6, TNF-𝛼, mRNA expressions of IL-1𝛽, IL-6, TNF-𝛼, Keap1, iNOS and COX-2 were decreased, and NQO1, Nrf2, and HO-1 were increased. In protein expression, JNK and Keap1 were decreased, NQO1, Nrf2 and HO-1 were increased, and no relationships were observed with the ERK and P38 by LCVC. Conclusions: These results suggest that LCVC may offer protective effects against LPS-induced inflammatory and oxidative responses through attenuating Nrf2/HO-1 pathway and MAPKs pathway. Therefore, we propose that LCVC has anti-inflammatory and anti-oxidative activities that have therapeutic potential in the treatment of inflammatory and oxidative disorders caused by the over-activation of macrophages.

• 대한화장품학회지
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• 제42권4호
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• pp.311-320
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• 2016

카바(Piper methysticum, P. methysticum)는 비뇨생식기 질환, 류머티즘, 위장 장애, 호흡기 자극 및 폐 통증에 대해 전통적으로 사용되는 것으로 알려져 있다. 본 연구에서는 카바에서 분리된 flavokavain C (FKC)가 염증성 유전자의 발현에 관여하는 전사인자인 핵요소-${\kappa}B$ (NF-${\kappa}B$) 의존성 산화 질소(NO) 생산 및 산화 질소 합성 효소(iNOS)의 발현에 리포폴리사카라이드(LPS) 처리된 대식세포에서 항 염증 활성을 나타낸다는 것을 확인하였다. FKC는 과산화수소와 같은 반응성 산소 종(ROS)의 축적을 억제하고, LPS로 유도된 NO 생성 및 각종 염증 관련 유전자(iNOS, IL-$1{\beta}$, IL-6)의 발현을 NF-${\kappa}B$ 및 MAPKs (ERK 및 JNK)의 억제를 통해 농도 의존적으로 줄일 수 있었다. 결론적으로, 이러한 결과는 FKC가 NF-${\kappa}B$ 경로와 MAPKs 포함한 염증 프로세스를 억제하는 능력을 가지고 있음을 나타내며, 이는 항 염증 및 항산화 효능 기반 기능성 화장품에 적용될 수 있음을 암시한다.