• 생명과학회지
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• 제21권10호
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• pp.1377-1384
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• 2011

Astaxanthin (ATX)은 다양한 생명체에서 생성되는 카로티노이드 색소이다. 본 연구에서는 ATX가 RAW264.7 cell에서 LPS에 의한 inducible nitric oxide synthase (iNOS), nitric oxide (NO), 염증성 사이토카인, nuclear factor-kappa B (NF-${\kappa}B$)와 reactive oxygen species (ROS)의 생성을 억제 시키는 지 또한, superoxide radical 소거능이 있는 지를 조사하였다. iNOS와 NF-${\kappa}B$는 immunoblot analysis로, interleukin (IL)-6와 tumour necrosis factor-${\alpha}$ (TNF-${\alpha}$)는 ELISA 법으로 분석하였다. NO 양은 nitrite의 양을 측정하였고, ROS는 2',7'-dichlorodihydrofluorescin diacetate (DCFH-DA) 법으로 superoxide radical 소거능은 superoxide radical scavenging activity assay로 검증하였다. 100 ${\mu}M$의 ATX 농도에서 LPS로 유도된 NO, IL-6 및 TNF-${\alpha}$ 같은 염증성 사이토카인의 생성 뿐만 아니라 iNOS 및 NF-${\kappa}B$의 발현도 억제되었다. 특히, IL-6 및 TNF-${\alpha}$ 생성에 있어 ATX의 최대 억제율은 각각 65.2% 및 21.2% 이었으며 LPS로 유도된 NF-${\kappa}B$의 전사활성을 억제하였다. 이러한 현상은 세포질에서 핵으로 NF-${\kappa}B$의 전위를 억제하는 것과 관련이 있다. 또한, 25-100 ${\mu}M$의 ATX 농도에서 세포 내 ROS 생성을 억제하였으며, 5 mg/ml 농도의 ATX는 동일농도의 ${\alpha}$-tocopherol에 비해 superoxide radical 소거능이 1.33배 높았다. 이러한 결과들은 ATX가 대식세포에서 ROS 생성 및 NF-${\kappa}B$ 활성을 저해하므로 iNOS의 발현, NO 및 염증성 사이토카인의 생성을 억제하며, 또한 우수한 superoxide radical 소거능을 보유한다는 것을 나타내었다. 결론적으로, ATX가 항염증제 및 항산화제로서 유용하게 사용될 수 있을 것으로 사료된다.

• 한방안이비인후피부과학회지
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• 제26권4호
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• pp.70-80
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• 2013

Objectives : Haliotidis Concha has been used to treat various human diseases such as liver dysfunction and inflammatory disorder. Although it has been shown the effects of Haliotidis Concha on the various diseases, it has almost not been studied about the anti-inflammatory effects of the Haliotidis Concha and its mechanisms. Methods : This research investigated the effects of the Haliotidis Concha ethanol extract (HCE) on the production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) as well as tumor necrosis factor-alpha (TNF-${\alpha}$). The protein expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were assayed by immunoblot analyses, and the productions of NO, $PGE_2$ and TNF-${\alpha}$ were assessed by ELISA. Results : Haliotidis Concha decreased the production of NO and $PGE_2$, and inhibited the expression iNOS and COX-2 proteins in a concentration-dependent manner in LPS-treated Raw 264.7 cells. HCE suppressed the ability of LPS to activate the signaling pathways of nuclear factor kappa B (NF-${\kappa}B$) as indicated by HCE inhibited nuclear NF-${\kappa}B$ level and I-${\kappa}B{\alpha}$ phosphorylation. Also, HCE inhibited mitogen-activated protein kinases (MAPKs). Conclusions : HCE repressed the production of LPS-inducible NO, $PGE_2$ and TNF-${\alpha}$, which may be mediated by inhibition of NF-${\kappa}B$ translocation. This study suggest the use for the treatment of acute inflammatory disorders.

• Tuberculosis and Respiratory Diseases
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• 제42권4호
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• pp.447-454
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• 1995

Cytokine release from alveolar macrophages and subsequent interaction of these cytokines with the bronchial epithelium can induce epithelial cells to release inflammatory mediators. Nitric oxide(NO), a highly reactive gas formed from arginine by nitric oxide synthase(NOS), is known to be involved in inflammation and edema formation, and the inducible form of NOS(iNOS) can be increased by cytokines. In this context, we hypothesized that lung epithelial cells could be stimulated by cytokines released by alveolar macrophages to express iNOS. To test this hypothesis, the murine lung epithelial cell line, LA-4, or the human lung epithelial cell line, A549, were stimulated with culture supernatant fluids from alveolar macrophages. NO production was assessed by evaluating the culture supernatant fluids for nitrite and nitrate, the stable end products of NO. Both murine and human cell culture supernatant fluids demonstrated an increase in nitrite and nitrate which were time- and dose-dependent and attenuated by $TNF{\alpha}$ and IL-$1{\beta}$ antibodies(p<0.05, all comparisons). Consistent with these observations, cytomix a combination of $TNF{\alpha}$, IL-$1{\beta}$, and $\gamma$-interferon, stimulated the lung epithelial cell lines as well as primary cultures of human bronchial epithelial cells to increase their NO production as evidenced by an increase in nitrite and nitrate in their culture supernatant fluids, an increase in the iNOS staining by immunocytochemistry, and an increase in iNOS mRNA by Northern blottin(p<0.05, all comparisons). The cytokine effects on iNOS were all attenuated by dexamethasone. To determine if these in vitro observations are reflected in vivo, exhaled NO was measured and found to be increased in asthmatics not receiving corticosteroids. These data demonstrate that alveolar macrophage derived cytokines increase iNOS expression in lung epithelial cells and that these in vitro observations are mirrored by increased exhaled NO levels in asthmatics. Increased NO in the lung may contribute to edema formation and airway narrowing.

• 한국약용작물학회지
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• 제15권5호
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• pp.324-328
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• 2007

To search for immunoactive natural products exerting anti-inflammatory activity, we have evaluated the effects of the ethanol extracts of Rubus coreanus Miq. (ERC) on lipopolysaccharide-induced nitric oxide (NO), tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$, and $Interferon-{\gamma}\;(IFN-{\gamma})$ production by RAW 264.7 macrophage cell line. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased $IFN-{\gamma}\;and\;TNF-{\alpha}$ production. Consistent with these results, the protein level of inducible Nitric Oxide Synthase (iNOS) and cyclooxygenase-2 (COX-2) was inhibited by ethanol extracts of ERC in a dose-dependent manner. These results suggest that ERC may exert anti-inflammatory and analgesic effects possibly by suppressing the inducible NO synthase and COX-2 expressions.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.385-393
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• 2008

As an attempt to search for bioactive natural products exerting anti-inflammatory activity, we evaluated the effects of the methanol extract of Polytrichum commune Hedw (PCM) (Polytrichaceae) on lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines release in murine macrophage cell line RAW 264.7. PCM potently inhibits the production of NO, $PGE_2$, tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Consistent with these results, PCM also concentration-dependently inhibited LPS-induced inducible NO synthase (iNOS) and cyclooxygase (COX)-2 at the protein levels, and iNOS, COX-2, TNF-$\alpha$ and IL-6 at the mRNA levels without an appreciable cytotoxic effect on RAW 264.7 macrophag cells. Furthermore, PCM inhibited LPS-induced nuclear factor-kappa B (NF-$\kappa$B) activation as determined by NF-$\kappa$B reporter gene assay, and this inhibition was associated with a decrease in the nuclear translocation of p65 and p50 NF-$\kappa$B. Taken together, these results suggest that PCM may play an anti-inflammatory role in LPS-stimulated RAW 264.7 macrophages through the inhibitory regulation of iNOS, COX-2, TNF-$\alpha$ and IL-6 via NF-$\kappa$B inactivation.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.92-103
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• 2009

Objectives : The present study was examined to evaluate the anti-inflammatory effects of the Humulus japonicus MeOH extracts (HJE) in vivo. Methods : The effects of HJE on anti-inflammation were measured by production of NO, iNOS (inducible Nitric Oxide Synthase), COX-2, I$\kappa$B$\alpha$ (Inhibitor kappa B alpha), NF$\kappa$B (Nuclear Factor kappa B), TNF-$\alpha$ (Tumor Necrosis Factor-alpha) and IL-1$\beta$ (Interleukin-1$\beta$), IL-6 in Raw 264.7 macrophage cells stimulated with LPS. Results : 1. All concentrations of HJE(0.03 and 0.10 mg/ml) had no significant cytotoxicity in Raw 264.7 cell during the entire experimental period. 2. The level of NO and iNOS in culture medium was dramatically increased by LPS application. However, these increases were dose-dependently(0.03 and 0.10 mg/ml) attenuated by treatment with HJE. 3. HJE extract reduced PGE2 levels in a dose-dependent manner as a consequence of inhibition of COX-2 protein expression in Raw 264.7 macrophage cells stimulated with LPS. 4. 0.10 mg/ml HJE significantly inhibited the phosphorylation of I$\kappa$B$\alpha$ indicating the suppression of NF-$\kappa$B pathway in Raw 264.7 macrophage cells stimulated with LPS. 5. 0.10 mg/ml HJE significantly inhibited the production of TNF-$\alpha$ in Raw 264.7 macrophage cells stimulated with LPS. 6. All concentrations of HJE significantly inhibited the production of IL-1$\beta$, IL-6 in Raw 264.7 macrophage cells stimulated with LPS. Conclusions : These results provide evidences that therapeutic effect of HJE on heat syndrome, especially due to the acute inflammation, are partly due to the reduction of some of inflammatory factors by inhibiting iNOS and COX-2 through the suppression of p-I$\kappa$B$\alpha$. Moreover, it suggests that the mechanism of action of HJE comes from the suppression of inflammatory mediators, such as NO, PGE$_2$ and pro-inflammatory cytokines.

• 생약학회지
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• 제38권4호
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• pp.339-348
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• 2007

This study were designed to evaluate the anti-inflammatory effects of $genistein-4'-O-{\alpha}-L-rhamnopyranosyl-(1-2)-{\beta}-D-glucopyranoside$ (GRG) isolated from Sophora japonica (Leguminosae) on the lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin ($PGE_2$) production by RAW 264.7 cell line. GRG significantly inhibited the LPS-induced NO and $PGE_2$ production. Consistent with these observations, GRG 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 GRG. Moreover, GRG 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 GRG are achieved by the downregulation of $NF-{\kappa}B$ activity, and that is also responsible for its anti-inflammatory effects.

• 대한한의학회지
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• 제29권1호
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• pp.47-59
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• 2008

Objective : Anti-inflammatory effects of the extract of Lycii Fructus on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells were investigated. Method : In order to assess the cytotoxic effect of Lycii Fructus on the raw 264.7 macrophages 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was performed. Reverse transcription-polymerase chain reaction(RT-PCR) analysis of the mRNA levels of tumor necrosis factor-$\alpha$(TNF-$\alpha$) and inducible nitric oxide synthase(iNOS) was performed in order to provide an estimate of the relative level of expression of these genes. The protein level of the inhibitor of nuclear factor-${\kappa}B(I{\kappa}B)$ and nuclear factor-${\kappa}B$(NF-${\kappa}B$) activity was investigated by Western blot assay. NO production was investigated by NO detection. Result : Lycii Fructus suppressed NO production by inhibiting the LPS-induced expressions of iNOS and TNF-$^-\alpha$ mRNA and iNOS protein in RAW 264.7 macrophage cells. Also, Lycii Fructus suppressed activation of NF-${\kappa}B$ in the nucleus. Conclusion : These results show that the extract of Lycii Fructus has anti-inflammatory effect probably by suppressing iNOS expressions through the down-regulation of NF-${\kappa}B$ binding activity.

• Natural Product Sciences
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• 제15권2호
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• pp.83-89
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• 2009

Polygoni multiflori Ramulus (PM), the stem of Polygonum multiflorum Thunb. has been widely used as a traditional medicine for the treatment of lots of diseases. In macrophages, nitric oxide is released as an inflammatory mediator and has been proposed to be an important modulator of many pathophysiological conditions in inflammation. In the present study, it was investigated that the inhibitory effects on NO and proinflammatory cytokines such as tumor necrosis factor alpha (TNF-${\alpha}$), interleukin-6 (IL-6) and the mechanism of down-regulation of immune response by 85% methanol extracts of PM in mouse (C57BL/6) peritoneal macrophages. Extracts of PM (0.1, 1 mg/ml) suppressed NO production and showed inhibition of proinflammatory cytokines like TNF-${\alpha}$, IL-6 and it attenuated iNOS and COX-2 expression via down-regulation of NF-${\kappa}$B activation. The present results indicate that the 85% methanol extracts of PM has an inhibitory effect on the production of NO through down-regulation of iNOS expression in LPS stimulated mouse peritoneal macrophages and therefore may be beneficial in diseases which related to macrophage-mediated inflammatory disorders.

• 동의생리병리학회지
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• 제27권1호
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• pp.78-82
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• 2013

It has been reported that Scolopendra morsitans L.(SML) has beneficial effects on human health and diverse diseases. The purpose of this study was to investigate the anti-inflammatory effects of ether extract from Scolopendra morsitans L. on lipopolysaccharide(LPS)-induced inflammatory response. Thus, we examined the inhibitory effect of SML ether fraction on LPS-induced increase of inflammatory mediators(NO, iNOS, COX-2, and $I{\kappa}B{\alpha}$) and pro-inflammatory cytokines(TNF-${\alpha}$) in RAW 264.7 cells. In the present study, SML ether extract itself decreased cell viability in a dose dependent manner(> 100 ${\mu}g/ml$). In addition, LPS increased NO production, iNOS expression and phosphorylation of $I{\kappa}-B{\alpha}$, which were blocked by the treatment of SML ether fraction in a dose dependent manner. Furthermore, the treatment of LPS increased TNF-${\alpha}$ production. However, the pretreatment of SML ether fraction prevented the LPS-induced TNF-${\alpha}$ production in dose dependant manner. Taken together, our results suggest that SML may be a beneficial drug against inflammatory diseases such as sepsis.