• 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.

• 대한본초학회지
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• 제34권1호
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• pp.51-57
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• 2019

Objectives : Nypa fruticans Wurmb. (NF) have been used as a traditional medicine to treat inflammatory diseases in East-South Asia. However, it is largely undiscovered whether NF water extract could exhibit anti-inflammatory activities against tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory responses on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the anti-inflammatory activity of NF water extract on TNF-${\alpha}$-induced inflammatory responses in HaCaT cells. Methods : To investigate the anti-inflammatory activites of NF water extract in HaCaT cells, the inflammatory model of HaCaT cells was established under a suitable concentration (10 ng/ml) of human TNF-${\alpha}$ (hTNF-${\alpha}$). HaCaT keratinocyte cells were pre-treated with NF water extract for 1 h, and then stimulated with hTNF-${\alpha}$. Then, the cells were harvested to measure the inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$), and pro-inflammatory cytokine including TNF-${\alpha}$ and interleukin (IL)-6. In addition, we examined the inhibitory mechanisms of NF, mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha ($I{\kappa}-B{\alpha}$) Results : The treatment of NF inhibited the hTNF-${\alpha}$-induced elevation of iNOS, COX-2, and $PGE_2$ in HaCaT cells. In addition, NF treatment inhibited the hTNF-${\alpha}$-induced elevation of TNF-${\alpha}$ and IL-6. Furthermore, NF treatment inhibited the activation of MAPKs but not degradation of $I{\kappa}-B{\alpha}$. Conclusions : Taken together, our result suggest that treatment of NF could inhibit the hTNF-${\alpha}$-induced inflammatory responses via deactivation of MAPKs in HaCaT cells. This study could suggest that NF could be a beneficial agent to prevent skin damage or inflammation.

• Journal of Applied Biological Chemistry
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• 제53권3호
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• pp.147-153
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• 2010

$\beta$-glycyrrhetinic acid (GA), the active principle of licorice (Glycyrrhiza glabra L.) has been reported to exhibit anti-inflammatory properties in different animal models. In this study, the effects of GA on the production of inflammatory mediators including tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-6, nitric oxide (NO), and prostaglandin E (pGE)-2 were examined in RAW 264.7 cells in vitro. Furthermore, to elucidate a possible mechanism for the inhibitory effect of GA on the production of TNF-$\alpha$, it was investigated whether the treatment of GA affects the I-${\kappa}B{\alpha}$ degradation and subsequent nuclear translocation of NF-${\kappa}B$. Various inflammatory responses were induced in the culture system by treating with a lipopolysaccharide (LPS). GA showed anti-inflammatory activities in dose-dependant manner with $IC_{50}$ of $5.4{\mu}M$ by inhibiting the production of TNF-$\alpha$ in RAW 264.7 cells. In addition, the treatment of GA blocked both I-${\kappa}B{\alpha}$ degradation and the nuclear translocation of NF-${\kappa}B$ from cytosol to nucleus. However, it did not affect the production of IL-6, NO, and PGE-2, implying the direct blocking of the production of TNF-$\alpha$ resulting from both the I-${\kappa}B{\alpha}$ degradation and the nuclear translocation of NF-${\kappa}B$. This finding might provide the underlying mechanism to explain the reported anti-inflammatory activities of GA in animal models.

• 약학회지
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• 제45권6호
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• pp.715-723
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• 2001

Indomethacin is well known as a prostaglandin (PG) E$_2$ synthetase inhibitor which has antipyretic and anti-inflammatory effects and reduces the risk of cancer Growing tumors greatly induce hypersensitive responses to lipopolysaccharide (LPS). Thus, this study was investigated the effect of indomethacin on the LPS-induced production of cytokines in sarcoma-bearing ICR mice. Indomethacin at doses of 5mg/kg was administered orally 30 minutes before i.p. injection of LPS (8 mg/kg) 5 times for 7 days. LPS remarkedly increased tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-1$\beta$, levels in both serum and splenic supernatants compared with those in controls, while indomethacin significantly reduced the LPS-increased levels of IL-1$\beta$, in both serum and supernatants. LPS significantly enhanced IL-2 levels in serum and interferon (IFN)-${\gamma}$ levels in supernatants, whereas indomethacin did not affect the LPS-increased levels of IL-2 and IFN-${\gamma}$. These data, therefore, indicate that indomethacin may attenuate the pathogenesis of IL-1$\beta$, induced by LPS and maintain the tumoricidal cellular immune effects by LPS-increased production of IL- 2 and IFN-${\gamma}$ in tumor-bearing state.

• 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.

• Natural Product Sciences
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• 제12권1호
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• pp.38-43
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• 2006

Ilekudinol B is one of the flavonoids isolated from Weigela subsessilis (Caprifoliaceae). In the present study, the suppression effect of ilekudinol B on tumor necrosis factor $(TNF)-{\alpha}-induced$ cyclooxygenase-2 (COX-2) expression was investigated in human colon epithelial cell line HT-29. Interleukin-8 (IL-8) production and prostaglandin $E_2\;(PGE_2)$ secretion was measured by enzyme-linked immunosorbent assay (ELISA). COX-2 and nuclear factor $(NF)-{\kappa}B$ expression were determined by Western blot analysis. Ilekudinol B significantly inhibited $TNF-{\alpha}-induced$ secretion of IL-8 and prostaglandin $E_2\;(PGE_2)$ from the human colon epithelial cell line HT-29 in a concentration-dependent manner. In addition, ilekudinol B remarkably diminished $TNF-{\alpha}-induced$ COX-2 expression and $NF-{\kappa}B$ p65 subunit translocation to the nucleus. In conclusion, our results indicate that ilekudinol B may have anti-inflammatory activity on $TNF-{\alpha}-dependent$ colonic inflammation.

• 한국균학회지
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• 제45권4호
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• pp.336-344
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• 2017

본 연구에서는 우리나라 주요 산과 섬에서 분리한 비병원성 야생효모들 중 항염 효과가 우수했던 Meyerozyma guilliermondii YJ34-2와 Rhodotorula graminis YJ36-1의 무세포 추출물들을 제조하여 대식세포 계열 RAW 264.7 세포에 대한 이들의 NO 생성 저해활성과 세포독성을 조사하였다. NO 생성 저해활성은 농도 의존적으로 높아 M. guilliermondii YJ34-2와 R. graminis YJ36-1 무세포 추출물을 1,000 mg/mL 처리 시 각각 51.6%와 81.4%를 보여 가장 높았고 RAW 264.7 세포에 대한 세포 생존율도 1,000 mg/mL 처리시 각각 88.4% (${\pm}3.1$)와 77.1% (${\pm}0.3$)로 가장 높았다. 두 효모들의 무세포 추출물 처리에 따른 prostaglandin $E_2$ 생성량은 농도 의존적으로 감소하여 각각의 무세포 추출물을 1,000 mg/mL 처리했을 때, tumor necrosis factor $(TNF)-{\alpha}$ 생성량이 59.2 (${\pm}43.1$), 73.2 (${\pm}38.1$)%로 감소하였고 prostaglandin $E_2$의 생성량도 52.8 (${\pm}1.9$), 71.2 (${\pm}3.7$)%로 감소하여 이 두 효모들의 항균활성을 검증할 수 있었다. 두 효모들의 NO 생성 저해물질 최적 생산조건을 조사한 결과 M. guilliermondii YJ34-2를 yeast extract-peptone- dextrose (YPD) 배지에 접종하여 $30^{\circ}C$에서 24시간 배양하여 얻은 무세포 추출물이 가장 높은 51.6 (${\pm}0.3$)%의 NO 생성 저해율을 보였고 R. graminis YJ36-1를 YPD 배지에 접종하여 $25^{\circ}C$에서 24시간 배양하였을 때 81.4 (${\pm}1.3$)%의 가장 높은 NO 생성 저해활성을 보였다.

• 대한임상검사과학회지
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• 제48권3호
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• pp.176-182
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• 2016

대식세포에서 염증반응이 진행되면, interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) 등의 cytokine들이 발현되어 inducible nitric oxide synthase (iNOS), prostaglandin E2(PGE2) 등의 염증유발인자가 생성된다. 오가피, 우슬, 두충 각각의 추출물이 어느 정도의 항 염증 효능을 보이며 어떤 pro-inflammatory cytokine의 발현을 억제하는지에 대한 연구를 진행하였다. 오가피, 우슬, 두충은 물 추출하고 동결 건조시켰다. 각각의 추출물의 구성 성분들이 잘 추출되었는지 확인하기 위하여 지표물질인 acanthoside D, 20-hydroxyecdysone, pinoresinol diglucoside를 HPLC로 분석하였다. 항 염증 효능을 확인하기 위하여 lipopolysaccharide (LPS)로 RAW 264.7 세포주를 자극하여 염증 반응을 일으킨 상태에서 각각의 추출물을 농도 별로 처리하고 NO assay를 통해 항 염증 효능을 확인하였으며 real time PCR로 pro-inflammatory cytokine들의 발현량을 측정하였다. 결과적으로 각각의 추출물은 지표성분들이 검출되었으며 오가피와 우슬이 두충보다 NO assay에서 높은 활성을 보였다. Cytokine 발현량 측정에서는 오가피와 우슬은 iNOS와 IL-6의 발현을 억제하였고, 우슬은 TNF-${\alpha}$의 발현을 억제하였다. 우리나라는 전통적으로 약재를 조합하여 처방하여 왔다. 본 연구는 관절염에 전통적으로 사용해 오던 약재들이 어떤 기전에 의하여 항 염증 반응을 보이는지 확인하고 이들을 조합하여 사용하였을 때 어떤 근거에 의하여 시너지 효능을 보이는지 확인하였다.

• 한방안이비인후피부과학회지
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• 제18권1호
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• pp.183-198
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• 2005

This experimental study was performed to investigate the anti-inflammatory effects of Phellodendri Cortex(PC), Indigo Naturalis(IN). Alumen(AM), and Chinensis Galla(CG). These four prescriptions most frequently have been used in of oriental medicine for the external the theraphy of Gu-Chang. This study was done to evaluate the activity of superoxide dismutase(SOD) and the inhibitory effects of the formation of cyclooxygenase-2(COX-2), tumor necrosis $factor-{\alpha}(TNF-{\alpha}),\;interlenkin-1{\beta}(IL-1{\beta})$, prostaglandin-E2(PGE2) on the monocyte and neutrophil. The results were summerized as follows. 1. IN has no cytotoxicity but PC, AM, CG have a little cytotoxicity by the increase of concentrations. 2. PC, IN, AM and CG all inhibited the formation of COX-2 in the human neutrophil. 3. $1000{\mu}g/m{\ell}$ of PC increased the formation of SOD in the human monocyte. 4. $1000{\mu}g/m{\ell}$ of PC, $100{\mu}g/m{\ell}$ and $1000{\mu}g/m{\ell}$ of IN, $100{\mu}g/m{\ell}$ of CG inhibited the formation of $TNF-{\alpha}$ in the human monocyte. 5. $10{\mu}g/m{\ell}$ of PC, $1000{\mu}g/m{\ell}$ of IN inhibited the formation of $IL-1\;{\beta}$ in the human monocyte. 6. PC, IN, AM and CG all did not inhibit the production of PGE2 in the human monocyte In addition, the results show that PC, IN, AM and CG all have anti-inflammatory effects and can be used fer the external theraphy of Cu-Chang.

• 대한의생명과학회지
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• 제19권3호
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• pp.195-205
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• 2013

Obesity may cause metabolic syndrome and adult diseases. This study was undertaken to investigate the ameliorative or useful effects of beanleaves on inflammation and liver damage in obese rat models. Rats were divided into three groups: a control group (normal diet, n=6), a fat diet group (45%-fat diet, n=7), and a bean leaf group (45%-fat+Korean bean leaves diet, n=7). Body weights in the bean leaf group were lower than those of the fat group (P<0.05). Serum tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and prostaglandin $E_2$ ($PGE_2$) concentrations were lower in both the control and bean leaf groups than in the fat group (P<0.001). TNF-${\alpha}$ concentrations in the bean leaf group were slightly higher than in the control group but statistically significant (P<0.05). The bean leaf group histologically exhibited lower fatty degeneration, spotty necrosis, and leukocyte infiltrations in hepatic tissues than those of the fat group. In the homogenized liver tissues, the cyclooxygenase-2 (COX-2) gene was only expressed in the fat group. The gene expression levels of hepatic TNF-${\alpha}$, inducible nitric-oxide synthase, peroxiome proliferator-activated receptor-${\alpha}$ (PPAR-${\alpha}$), poly (ADP-ribose) polymerase (PARP), and transforming growth factor-${\beta}1$ (TGF-${\beta}1$) were weaker in the bean leaf group than in the fat group. These results suggest that adding bean-leaves to the diet may ameliorate obesity-induced systemic inflammation and liver damage and that bean leaves may be a useful food for preventing obesity and thereby metabolic syndrome and adult diseases.