• Applied Biological Chemistry
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• 제48권3호
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• pp.246-251
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• 2005

본 시험은 면역계에서 중요한 역할을 담당하는 세포가운데 하나인 대식세포(Raw 264.7)를 게르마늄 강화 효모가 활성화시키는지 여부를 알아보기 위하여 대식세포의 배양액으로부터 NO와 $TNF-{\alpha}$의 생성을 in vitro 상에서 알아보고 iNOS의 발현 정도를 확인하고자 실시하였으며, 그 결과는 다음과 같다. 게르마늄 강화 효모 처리 후 세포 생존율(%)과 NO 생성량은 $10\;{\mu}g/ml$의 처리 농도에서 유의적으로 증가하였다(p < 0.05). 또한 NO 생성을 매개하는 iNOS의 발현 역시 $10\;{\mu}g/ml$의 농도에서 증가한 것으로 나타났다. 면역 및 항암 조절 인자로 알려진 $TNF-{\alpha}$의 생성 역시 농도 의존적으로 증가하였으며, $10\;{\mu}g/ml$의 처리 농도에서 유의적으로 증가하였다(p < 0.05). 이는 게르마늄 강화 효모가 항암 및 면역 증진 기능과 관련이 있는 $TNF-{\alpha}$ 분비를 촉진시키는데 영향을 준 것으로 사료된다. 따라서 본 시험 모델에서 게르마늄 강화 효모는 iNOS 발현을 조절하여 NO 및 $TNF-{\alpha}$의 생성을 매개하여 면역조절 기능에 도움을 줄 것으로 사료된다. 이것은 면역기능에 있어서 중요한 역할을 하는 대식세포를 활성화시켜 면역력을 활성화시키므로 세포성 면역의 활성화를 도모하고 손상된 면역 체계의 정상화에 영향을 주어 면역 강화 및 항암 예방 기능성 신소재로의 가능성을 지니는 것으로 사료된다.

• Journal of Periodontal and Implant Science
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• 제34권2호
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• pp.461-474
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• 2004

The purpose of this study was to assess some biological activities of lipopolysaccharides (LPSs) from P. intermedia and P. nigrescens. LPS was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. $TNF-{\alpha}$ production was determined by enzyme-linked immunosorbent assay. Western blot analysis of iNOS and analysis of reverse transcription (RT)-PCR products were carried out. LPS from P. intermedia demonstrated higher KDO content than those from two stains of P. nigrescens. LPSs from P. intermedia and P. nigrescens were mitogenic for spleen cells of BALB/C mouse. The present study clearly shows that LPSs from P. intermedia and P. nigrescens fully induced iNOS expression and NO production in RAW264.7 cells in the absence of other stimuli. Moreover, LPSs from P. intermedia and P. nigrescens clearly induced $TNF-{\alpha}$ production in RAW264.7 cells. The biological activities of LPS from P. intermedia was found to be comparable to those of P. nigrescens LPS. The ability of LPSs from P. intermedia and P. nigrescens to promote the production of NO and $TNF-{\alpha}$ may be important in the pathogenesis of inflammatory periodontal disease.

• 약학회지
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• 제46권5호
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• pp.343-347
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• 2002

In the present study, effects of essential oils isolated from various plants have been evaluated on lipopolysaccharide (LPS)-induced release of nitric oxide (NO), prostaglandin E$_2$(PGE$_2$) and tumor necrosis factor-a (TNF-$\alpha$) by the macrophage RAW 264.7 cells. Among the tested essential oils, essential oil of Ligularia fischeri var. spiciformis (LF-oil) significantly inhibited the LPS-induced generation of NO, PGE$_2$ and TNF-$\alpha$ in Raw 264.7 cells. Consistent with these observations, the expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 enzyme was inhibited by LF-oil in a concentration-dependent manner. Thus, this study suggests that inhibition of release of iNOS, COX-2 expression, and TNF-$\alpha$ by the essential oil of Ligularia fischer may be one of the mechanisms responsible for the anti-inflammatory effects of this medicinal plant.

• 한국자원식물학회지
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• 제25권3호
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• pp.299-307
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• 2012

This work aimed to elucidate the anti-inflammatory effects of ethyl acetate fraction from Cnidium officinale Makino with a cellular system of LPS-stimulated RAW 264.7 and THP-1 cells. Some key pro-inflammatory cytokines and mediators including NO, iNOS, $PGE_2$, COX-2, TNF-${\alpha}$, NF-${\kappa}B$ p50 and NF-${\kappa}B$ p65 were studied by sandwich ELISA and western blot analysis. Ethyl acetate fraction could significantly inhibit the production of NO, $PGE_2$, TNF-${\alpha}$, iNOS and COX-2 in LPS-stimulated cell than that of single LPS-stimulated. And ethyl acetate fraction suppresses the activation of NF-${\kappa}B$ p50 and NF-${\kappa}B$ p65. All the results showed that ethyl acetate fraction had a good anti-inflammatory effect on LPS-stimulated RAW264.7 and THP-1 cells. Taken together, the anti-inflammatory actions of ethyl acetate fraction from Cnidium officinale Makino might be due to the down-regulation of NO, $PGE_2$, TNF-${\alpha}$, iNOS and COX-2 via the suppression of NF-${\kappa}B$ activation.

• 동의생리병리학회지
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• 제31권4호
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• pp.220-225
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• 2017

Lithospermi Radix (LR) is known to have an anti-inflammatory effect. However, the mechanisms are not well known. In this study, LPS-induced mouse RAW 264.7 macrophage cells were treated with LR to investigate the time-dependent inflammation response of LR. RAW 264.7 cells were treated with various concentrations of LR for 24 hours, followed by MTS assay. Cell viability was increased at all experimental concentrations. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by treatment of RAW 264.7 cells with LR at a concentration of $200{\mu}g/ml$ for 6 hours and 24 hours. Treatment of LR with $200{\mu}g/ml$ concentration for 6 hours promoted mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS in LPS-induced RAW 264.7 cells. However, $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS mRNA expression was suppressed by treatment of LR with $200{\mu}g/ml$ concentration for 24 hours in LPS-induced RAW 264.7 cells. These results suggest that the effect on inflammation of LR is promptly promoted and then to rapidly alleviate the inflammatory reaction. This study proposes that the time-dependent activities of herbal medicine is a very important factor in analyzing the anti-inflammatory effect of various herbal medicines including LR.

• 대한화장품학회지
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• 제40권1호
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• pp.45-54
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• 2014

각질형성세포주인 HaCaT세포에서 Lipopolysaccharide (LPS)로 유도하는 염증반응에는 tumor neurosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-1${\alpha}$ (IL-1${\alpha}$), interleukin-6 (IL-6), 및 interleukin-8 (IL-8)와 같은 염증 유발 사이토카인(pro-inflammatory cytokines)이 증가되고, 또한 염증반응을 매개하는 inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) 및 prostaglandins E2 (PGE2)도 증가된다. 본 연구에서는 이렇게 유도된 염증 반응에서 효과적으로 염증 관련인자를 조절할 수 있는 천연물을 발굴하고자 하였다. 그 결과, 마가목, 쐐기풀, 죽여 및 오배자 혼합 추출물이 IL-1${\alpha}$, TNF-${\alpha}$, IL-6 및 IL-8의 발현을 억제하고 COX-2, iNOS 및 PGE2의 발현도 억제함을 확인하였다. 아울러 IQ chamber$^{(R)}$을 이용하여 피부에 첩포할 때 sodium lauryl sulfate (SLS)로 유발한 피부자극에서 이들 혼합물이 효과적으로 그 자극을 억제함을 확인하였다. 따라서 본 연구 결과로부터 마가목, 쐐기풀, 죽여 및 오배자의 혼합물이 피부자극완화용 화장품 소재로서 민감성 피부 등을 위한 제품에 유용하게 사용될 수 있음을 확인하였다.

• 한국식품영양과학회지
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• 제39권8호
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• pp.1097-1101
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• 2010

본 연구의 목적은 일상생활에서 쉽게 섭취할 수 있는 당근, 무, 무청, 우엉, 표고버섯을 재료로 한 야채 스프의 항염증성을 검토하기 위한 것으로, RAW264.7 세포에서 LPS에 의해 유도되는 염증물질 중 NO와 염증성 사이토카인 TNF-$\alpha$에 대한 억제력을 관찰하였다. 실험결과 실험에 사용된 야채스프는 세포독성을 나타내지 않았다. 세포독성이 없음을 확인 한 후 LPS로 처리한 RAW264.7 세포에서 야채스프의 NO 생성억제를 평가한 결과 RAW264.7 세포만 배양한 대조군에서 NO의 농도는 매우 낮게 측정되었으며, LPS를 처리한군에서 NO의 농도는 현저히 증가되었다. 또한 야채스프를 처리한 실험군은 NO 생성이 유의성 있게 억제되었으며, RAW264.7 세포를 LPS로 처리 시 iNOS가 발현되어 NO를 생성하게 되므로 iNOS의 발현에 미치는 영향을 평가하였다. LPS 처리 시 iNOS 유전자 발현이 유의하게 증가되었으나, 야채스프를 전 처리한 실험군에서 iNOS의 양이 감소하였다. 야채스프에 의한 iNOS의 발현억제는 NO 생성억제를 유도를 의미한다. Proinflammatory cytokines인 TNF-$\alpha$ 분비 억제를 살펴본 결과 LPS를 처리한 군에서 TNF-$\alpha$의 분비가 현저하게 증가되었으나 야채스프를 처리한 군에서는 유의한 억제를 나타냈다. 이상과 같이 야채스프는 RAW264.7 세포에서 LPS에 의해 유도되는 NO의 생성억제, iNOS의 발현을 억제시켰으며, proinflammatory cytokines인 TNF-$\alpha$의 분비량도 억제시켰다. 이는 야채수프의 항염증성을 입증하기 위한 기초자료로 활용이 가능하다고 사료된다.

• 대한한의학회지
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• 제24권2호
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• pp.138-147
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• 2003

Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

• 약학회지
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• 제55권6호
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• pp.456-461
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• 2011

The aim of the present study is to investigate the anti-inflammatory effect of a Rice Bran Ethanol Extract (RBE). Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the RBE on pro-inflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of RBE, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the RBE reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the RBE may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory material.

• 융합정보논문지
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• 제9권6호
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• pp.218-226
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• 2019

본 연구의 목적은 삼내자(Kaempferia Galanga) 열수추출물의 항산화, 세포독성, 항염 효능을 조사하여 화장품 성분으로서 활용 가능성을 알아보고자 하였다. 항산화 활성 평가를 위해 DPPH, ABTS 라디컬 소거능 활성, FRAP 환원력, 총폴리페놀 함량을 측정하였다. 쥐의 대식세포인 RAW264.7 세포를 대상으로 MTT assay를 수행하여 세포독성을 확인하였다. 항염 효능을 알아보기 위해 LPS로 유도된 RAW264.7 세포에서 nitric oxide(NO), $TNF-{\alpha}$ 분비 및 iNOS, $TNF-{\alpha}$ mRNA 발현 수준을 조사하였다. 그 결과 DPPH와 ABTS 라디칼 소거능은 농도 의존적으로 라디칼 소거 활성이 증가하였다. FRAP 값은 5 mg/mL에서 24.5 uM로 가장 높게 나타났으며, 총폴리페놀 함량은 $1.28{\pm}0.064mg\;GAE/g$로 나타났다. KG 추출물 농도 0.625~2.5 mg/mL에서 세포독성은 보이지 않았다. 또한 RAW264.7 세포에서 LPS 유도에 의한 NO, $TNF-{\alpha}$ 분비 그리고 iNOS, $TNF-{\alpha}$의 mRNA 발현이 KG 추출물에 의해 유의하게 감소하였다. 이러한 결과는 삼내자 열수추출물이 안전하고 효과적인 화장품 원료로서의 활용 가능성이 있음을 보여준다.