• 동의생리병리학회지
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• 제19권5호
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• pp.1405-1410
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• 2005

Asthma is recognized today as an inflammatory disease of the lung characterized by acute non-specific airway hypersensitiveness in association with chronic pulmonary inflammation. Gamijihwang-tang(GJT), a fortified prescription of YMJHT, is applied for the treatments of chronic coughing and asthma, and post-delivery coughing and asthma in the gynecology. Also in the clinical practice, GJT is known to be very effective for controlling coughing and asthma as a cold sequoia. In this study, we investigated the effects of GJT on the lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production, and on the level of inducible nitric oxide synthase (iNOS) and Cyclooxygenase-2 expression in murine macrophage RAW 264.7 cells. We found that GJT inhibited LPS-induced NO and $PGE_2$ production in a dose dependent manner. Furthermore, GJT inhibited the expression of LPS-induced iNOS and COX-2 protein and mRNA expression in RAW 264.7 macrophages. Treatment with GJT of RAW 264.7 cells transfected with a reporter construct indicated a reduced level of LPS-induced nuclear factor-KB (NF-kB) activity and effectively lowered NF-kB binding as measured by transient transfection assay. These results suggest that the main inhibitory mechanism of the GJT may be the reduction of iNOS and COX-2 gene expression through blocking of NF-kB activation.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.282-288
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• 2023

코로나19 바이러스의 대유행으로 바이러스와 같은 외부 병원균으로부터 우리의 몸을 보호하는 면역 기능 개선 건강기능식품의 시장은 점차 증가하고 있다. 우리는 본 연구에서 높은 조단백, 조지방, 식이섬유 함량을 나타내는 고영양식품인 청국장이 면역 강화 기능을 나타냄을 밝혀내고자 하였다. 청국장 열수 추출물은 RAW 264.7 세포에서 세포독성을 나타내지 않으며, 대식세포의 nitric oxide, reactive oxygen species 및 interleukin (IL)-6, IL-1β, tumor necrosis factor-α 사이토카인의 생산량을 증가시켰다. 또한, 청국장 열수 추출물은 RAW 264.7 세포에서 inducible nitric oxide synthase 및 cyclooxygenase-2의 발현을 유의적으로 증가시켰다. 청국장 열수 추출물은 RAW 264.7 세포에서 I kappa B kinase α/β와 I kappa B (IκB)α의 인산화 및 IκBα의 degradation을 증가시켰으며, Nuclear factor-kappa B p65의 인산화를 증가시켜 p65의 세포질에서 핵으로의 이동을 촉진하였다. 이러한 연구 결과는 청국장 추출물이 선천성 면역 반응을 강화하는데 유망한 건강기능식품 소재로 활용될 수 있음을 시사한다.

• 동의생리병리학회지
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• 제18권6호
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• pp.1622-1627
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• 2004

Nephrite has been widely used as a medicinal mineral resource to treat a numerous chronic diseases and to replenish vital essence and blood in the Korean traditional medicine. However, as of yet, there is little understanding of the pharmacological and biochemical mechanisms of its therapeutic effects as regards anti-inflammation. We therefore examined whether nephrite represses the expression of major inflammation mediators, such as interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), in LPS-stimulated murine macrophage cell line, RAW264.7 by using RT-PCR. The powder suspension and water extracts of nephrite significantly inhibited the mRNA expression of the mediators, despite a little toxic effects on growth of RAW 264.7 cells within the concentration range tested. These experimental results suggested that the nephrite can be utilized as a functional mineral exerting the anti-inflammation medicinal effect.

• 동의생리병리학회지
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• 제22권4호
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• pp.878-883
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• 2008

Conchiolin is a complex protein which is secreted by the mollusc's outer epithelium to be the organic basis of mollusc shell. This study is to investigate a potential anti-inflammatory activity of conchiolin of oyster shell (COS). We tested the effects of COS on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and prostaglandin E2 (PGE 2) in a murine macrophage cell line, RAW 264.7. COS inhibited production of NO and PGE2 in a dose dependent manner and also decreased the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and interleukin-6 (IL-6). These results suggest that COS can inhibit production of pro-inflammatory mediators and might be a useful source to treat inflammation.

• Journal of Microbiology
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• 제45권5호
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• pp.373-378
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• 2007

Based on observations that lactic acid bacteria have the ability to activate macrophages, we assessed the potential effects of eight different Lactobacillus strains treated with gastrointestinal enzymes on the production of nitric oxide and various cytokines in macrophages. RAW 264.7 murine macrophage cells were cultured with either precipitates or supernatants of Lactobacillus strains digested with pepsin followed by pancreatin. The increased production of nitric oxide and interleukin $(IL)-1{\beta}$, IL-6, IL-12 and tumour necrosis factor $(TNF)-{\alpha}$ were observed when cultured with precipitates, and this effect was largely strain-dependent. In contrast, the exposure of RAW 264.7 cells to supernatants produced weaker or nearly undetectable effects in comparison to the effects of exposure to precipitates. The induction of nitric oxide appeared to be unaffected. These results demonstrate that nitric oxide and cytokines were effectively induced when the bacterial precipitate was treated with macrophages. The results of the present study also indicate that Lactobacillus strains treated with digestive enzymes are capable of stimulating the production of nitric oxide and cytokines in macrophages, which may modulate the gastrointestinal immune function of the host when it is given as a feed additive.

• Journal of Applied Biological Chemistry
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• 제57권3호
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• pp.205-210
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• 2014

Persicaria orientalis (L.) Spach (Po)와 Potentilla fragarioides var. major Maxim (Pf)의 추출물로 reactive oxygen species (ROS)와 같은 free radical의 억제를 통한 항염증 활성을 살펴보았다. 또한, Po와 Pf로 LPS를 처리한 murine macrophage RAW 264.7 세포에서 발생한 nitric oxide의 저해와 NF-${\kappa}B$의 핵으로 translocation의 저해를 살펴보았다. 3-morpholinosydnonimine hydrochloride (SIN-1) $50{\mu}M$에 의해 유도된 ROS의 Po에 의한 50% 저해값 ($IC_{50}$)은 $23.35{\pm}1.27mg/mL$, Pf에 의한 $IC_{50}$은 $8.46{\pm}1.22mg/mL$이었다. 또한, SIN-1 $50{\mu}M$에 의해 유도된 peroxynitrite의 Po에 의한 $IC_{50}$은 $2.19{\pm}0.04mg/mL$, Pf에 의한 $IC_{50}$은 $0.80{\pm}0.02mg/mL$이었다. LPS 1 mg/mL을 처리한 RAW 264.7 세포에서 Nitric oxide는 증가하였으나 Po와 Pf 추출물을 처리한 그룹에서 농도의존적, 유의적으로 감소하였다. Po 추출물을 처리한 그룹의 Nitric oxide 생성량은 $13.34{\pm}0.67{\mu}M$, Pf 추출물을 처리한 그룹의 Nitric oxide 생성량은 $11.45{\pm}0.57{\mu}M$이었다. 또한, Po와 Pf 추출물은 LPS를 처리한 RAW 264.7 세포에서 NF-${\kappa}B$의 핵으로의 translocation을 저해하였다. 그러므로, Po와 Pf는 항염증 소재로서의 가능성이 충분하다고 사료된다.

• 한국자원식물학회지
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• 제31권3호
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• pp.202-210
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• 2018

Abeliophyllum distichum is a medicinal plant used in regional traditional medicine to relieve pain in inflammatory processes. In this study, anti-inflammatory effects of Abeliophyllum distichum flower (ADF) extract were examined. Furthermore, possible molecular mechanisms of the anti-inflammatory effects were dissected. The anti-inflammatory activity was investigated by inhibition of lipopolysaccharide (LPS) induced pro-inflammatory cytokine production in murine macrophage-like cell line Raw264.7 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK, and MAPK, and the nuclear expression of nuclear factor NF-${\kappa}B$ p65 were investigated by Western blot analysis. The extract of ADF significantly decreased the production of pro-inflammatory cytokines. In addition, the extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of NF-${\kappa}B$ p65 in activated cells. Our findings provide evidence for the popular use of Abeliophylli distichum in inflammation around Goesan region and also suggest that the flower extract has potential therapeutic benefits against various inflammatory diseases.

• 대한약침학회지
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• 제6권2호
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• pp.77-90
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• 2003

The purpose of this study was to investigate the effect of Bee Venom on the lipopolysaccharide(LPS), sodium nitroprusside(SNP), hydrogen peroxide($H_2O_2$)-induced expressions of cyclooxygenase-2(COX-2), p38, jun N-terminal Kinase(JNK) and extra-signal response kinase(ERK) in RAW 264.7 cells, a murine macrophage cell line. Method : The expressions of COX-2, p38, JNK and ERK were determined by western blotting with corresponding antibodies. Results : 1. The 0.5, 1 and $5\;{\mu}g/ml$ of bee venom inhibited significantly LPS and SNP-induced expression of COX-2 compared with control, respectively. The 0.5, 1 and $5\;{\mu}g/ml$ of bee venom inhibited insignificantly $H_2O_2$-induced expression of COX-2 compared with control, respectively. 2. The 0.5, 1 and $5\;{\mu}g/ml$ of bee venom inhibited significantly LPS, SNP and $H_2O_2$-induced expression of p38 compared with control, respectively. 3. The 1 and $5\;{\mu}g/ml$ of bee venom inhibited significantly SNP-induced expression of JNK compared with control, respectively. All of bee venom inhibited insignificantly LPS and $H_2O_2$-induced expression of JNK compared with control, respectively. 4. The $5\;{\mu}g/ml$ of bee venom inhibited significantly SNP-induced expression of ERK, the $0.5\;{\mu}g/ml$ of bee venom increased significantly $H_2O_2$-induced expression of ERK compared with control. The 0.5, 1 and $5\;{\mu}g/ml$ of bee venom inhibited insignificantly LPS-induced expression of ERK compared with control, respectively.

• Journal of Microbiology and Biotechnology
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• 제29권5호
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• pp.820-826
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• 2019

This study evaluated the anti-inflammatory potential of a grasshopper ketone (GK) isolated from the brown alga Sargassum fulvellum on lipopolysaccharide (LPS)-induced RAW 264.7 murine macrophage cell line. GK was isolated and purified from the n-hexane fraction and its structure was verified on the basis of NMR spectroscopic data. GK up to $100{\mu}g/ml$ is not cytotoxic to RAW 264.7, and is an effective inhibitor of LPS-induced NO production in RAW 264.7 cells. The production of pro-inflammatory cytokines, including IL-6, $IL-1{\beta}$, and $TNF-{\alpha}$ was found significantly reduced in $0.1-100{\mu}g/ml$ dose ranges of GK treatment (p < 0.05). We confirmed the dose-dependent and significant inhibition of iNOS and COX-2 proteins expression. In addition, it has been shown that GK induces anti-inflammatory effects by inhibiting MAPKs (ERK, JNK, and p38) and $NF-{\kappa}B$ p65 phosphorylation. Our results show that the anti-inflammatory properties of GK may be due to the inhibition of the $NF-{\kappa}B$ and MAPKs pathways, which are associated with the attenuation of cytokine secretion.

• Biomolecules & Therapeutics
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• 제8권1호
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• pp.73-77
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• 2000

Rheum undulatum L. has been used as Rhei Radix in Korean Pharmacopea although their pharmacological effects were not studied much. In this studym, we tested anti-inflammatory effect as a representative activity of Rheum undulatum L. extracts using cyclooxygenase (COX)-2 inhibition. Murine macrophage, Raw 264.7 cells were incubated with lipopolysaccharide (1 $\mu\textrm{g}$/ml) to induce COX-2. The prostagladin $E_2$ ($PGE_2$) levels as an indicator of COX-2 activity were determined in the culture medium using ELISA. Inhibition of acetylsalicylic acid (ASA) as a standard, aloe-emodin, chrysophanol, rhein, 80% ethanol extract of Rheum undulatum L. (Ex) and ether fraction (Fr) after acid hydrolysis of Rheum undulatum L. were tested in induced COX-2 described above. $IC_{50}$ values were 0.082 $\mu\textrm{g}$/ml for ASA. 181 $\mu\textrm{g}$/ml for aloe-emodin, 3.65 $\mu\textrm{g}$/ml for emodin, 144 $\mu\textrm{g}$/ml for chrysophanol, 39.8 $\mu\textrm{g}$/ml for rhein, 141 $\mu\textrm{g}$ of herb/ml for Ex, and 95.7 $\mu\textrm{g}$ of herb/ml for Fr. We found that Ex and Fr of Rheum undulatum L. were more effective than other anthraquinones, since their $IC_{50}$ are lower than others.