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Verification of Anti-inflammatory Activities of the Ethanol Extracts of Glechoma hederacea var. longituba in RAW 264.7 Cells (RAW 264.7 세포에서 긴병꽃풀 에탄올 추출물의 항염증 활성 검증)

  • Lee, Jin-Young;Yoo, Dan-Hee;Jeong, Yong-Seong;Joo, Sung-Hyun;Chae, Jung-Woo
    • Journal of Life Science
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    • v.28 no.4
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    • pp.429-434
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    • 2018
  • In this study, we investigated the potential of Glechoma hederacea var. longituba 70% ethanol extract as a natural functional material by examining the anti-inflammatory effect of it. Macrophages results in (Raw 264.7) induced by lipopolysaccharide (LPS). Confirming the viability of the macrophages Glechoma hederacea var. longituba 70% ethanol extract showed a 95.8% survival rate at $1,000{\mu}g/ml$ concentration. Anti-inflammatory activity was examined the inhibitory tests on the production of LPS included nitric oxide (NO) in RAW 264.7 cells by Griess assay. The result showed that NO production deterrent effect of 37.4% at a concentration of $1,000{\mu}g/ml$. The deterrent effect of GG 70% ethanol extract on protein expression of inducible NOS (iNOS) and Cyclooxygenase-2 (COX-2) was measured by Western blotting using the concentrations 50, 100 and $500{\mu}g/ml$, with ${\beta}$-actin used as the positive control. The inhibitory effect of iNOS and COX-2 mRNA expression was measured by reverse transcription-polymerase chain reaction (RT-PCR) using 50, 100 and $500{\mu}g/ml$ concentration of GG 70% ethanol extract, with GAPDH used as the positive control. In experiments using Western blot and RT-PCR when compared with the control group vitamin C it was confirmed that the 70% ethanol extract from GG suppressed. When compiling the results of this study, we confirmed the possibility of GG 70% ethanol extract as an anti-inflammatory material.

Antioxidant and Anti-inflammatory Effects of Taraxacum hallaisanense Nakai Extracts (좀민들레(Taraxacum hallaisanense Nakai) 추출물의 항산화 및 항염증 활성 효과)

  • Nan, Li;Choo, Byung-Kil
    • Korean Journal of Organic Agriculture
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    • v.26 no.3
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    • pp.501-514
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    • 2018
  • Taraxacum hallaisanense (pr), a species of the family Compositae is a perennial herb plants that inhabit to Jeju Island. In the study, we performed to determine the anti-inflammatory effects in LPS-induced RAW 264.7 cells and antioxidant activity of ethanol extracts from T. hallaisanense whole plants. The antioxidant activity of extracts was measured by contents of polyphenol and flavonoid, DPPH radical scavenging, and reducing power activity. The anti-inflammatory effect of T. hallaisanense extracts was measured by NO and $IL-1{\beta}$ production inhibitory activity and the expression of pro-inflammatory in lipopolysaccharide (LPS)-induced Raw 264.7 cells. Also, the expression of pro-inflammatory genes such as iNOS, COX-2 and NF-kB protein were reduced. In the cytotoxicity measurement by cytotoxicity kit, the extract was exhibited Raw 264.7 cell viabilities as nontoxic result in concentration of $25{\sim}400{\mu}g/ml$. These results indicated that ethanol extracts of T. hallaisanense whole plants expected development possibility as nutrial additives through high anti-inflammatory effects and antioxidant activity.

Inhibition of Nitric Oxide Production by Coumarins from Peucedanum japonicum in LPS-Activated RAW 264.7 cells (갯기름나물의 쿠마린에 의한 RAW 264.7 세포주의 Nitric Oxide 생성 저해활성)

  • Choi, Hee-Cheol;Rho, Tae-Cheol;Kim, Bo-Yeon;Ko, Hack-Ryong;Oh, Won-Keun;Seong, Chang-Keun;Mheen, Tae-Ick;Ahn, Jong-Seog;Lee, Hyun-Sun
    • Korean Journal of Pharmacognosy
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    • v.30 no.2
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    • pp.99-104
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    • 1999
  • During the screening for inhibitors of nitric oxide production in LPS-activated macrophage, RAW 264.7 cells. Five coumarins were isolated from chloroform extract of the root of Peucedanum japonicum. They were identified as praeruptorin A (1), xanthotoxin (2), psoralen (3), isopimpinellin (4), bergapten (5) on the basis of spectroscopic methods. The $IC_{50}$ values for nitrite production by activated macrophages were about $1.5\;{\mu}g/ml$ (1), $0.3\;{\mu}g/ml$ (2), $1.0\;{\mu}g/ml$ (3), $25\;{\mu}g/ml$ (4), $25\;{\mu}g/ml$ (5), respectively. However, the inducible nitric oxide synthase (iNOS) was not inhibited by treatment with these compounds. Their inhibitory effect on nitric oxide production was resulted from the supperssion of iNOS expression.

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The roles of ceramide on the cellular signal transduction in RAW 264.7 murine macrophages activated with lipopolysaccharide and interferon-gamma.

  • Park, Sung-Sik;Chun, Young-Jin
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.156.1-156.1
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    • 2003
  • Cerarmide acts as a lipid second messenger in the cellular signal transduction and is involved in mediating a variety of cell functions such as proliferation, differentiation, growth arrest, and apoptosis. In the present study, we have investigated the effect of ceramide on cellular cytotoxity and reactive oxygen species (ROS) to understand the relationship between them. Ceramide treatment significantly increased cell death in RAW 264.7 murine macrophages activated with lipopolysaccharide (LPS) and interferon-g (IFN-g). (omitted)

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Suppressive Effect of Chlorella Methanol Extract on Oxidative Stress and NFkB Activation in RAW 264.7 Macrophages

  • Park, Ji-Young;Lee, Hyo-Sun;Song, Young-Sun
    • Preventive Nutrition and Food Science
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    • v.8 no.2
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    • pp.124-129
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    • 2003
  • This study was designed to investigate whether a methanol extract of chlorella can suppress oxidative stress and nuclear factor kB (NFkB) activation in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. Treatment of RAW 264.7 cells with chlorella methanol extract (25, 50, and 100 $\mu$g/mL) significantly reduced LPS-stimulated nitric oxide production in a dose-dependent manner. Treatments with chlorella methanol extract at all concentrations also reduced thiobarbituric acid-reactive substances accumulation and enhanced glutathione level at 50 and 100 $\mu$g/mL levels. The specific DNA binding activities of NFkB on nuclear extracts in cells treated with 50 $\mu$g/mL and 100 $\mu$g/mL chlorella methanol extracts were significantly suppressed. These results suggest that chlorella methanol extract has mild antioxidative activity and the ability to suppress intracellular oxidative stress and NFkB activation.

Anti-oxidation and Anti-inflammatory Effect of Asiasari Radix in RAW 264.7 Cells (세신(細辛) 주정(酒錠) 추출물(抽出物)이 LPS로 유발된 RAW 264.7 Cell의 염증 및 항산화 반응에 미치는 영향)

  • Lee, Yu-Chen;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.3
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    • pp.99-110
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    • 2014
  • Objectives The purpose of this study was to investigate the Anti-oxidation and anti-inflammatory effects of ethanol extract from asiasari radix (AR) on lipopolysaccharide (LPS)-induced in RAW 264.7 Cells Methods Anti-oxidative effects of AR were measured by scavenging activities of 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and production of reactive oxygen species (ROS) in RAW 264.7 cells. Anti-inflammatory effects of AR were measured by mediators including nitric oxide(NO), interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), tumor necosis factors-${\alpha}$ (TNF-${\alpha}$) and iNOS, IL-$1{\beta}$, IL-6, TNF-${\alpha}$ mRNA expression in RAW 264.7 cells. Results Total phenolic content was expressed $28.77{\pm}1.67$. DPPH radical Scavenging was increased depend on AR ethanol extract. ABAT radical Scavenging was increased depend on AR ethanol extract. Production of ROS was significantly decreased by AR ethanol extract on concentration of 100 (${\mu}g/ml$). Production of NO was significantly decreased by AR ethanol extract on concentration of $100({\mu}g/ml)$. Production of IL-$1{\beta}$, interleukin-6 and TNF-${\alpha}$ were increased depend on AR ethanol extract. And Production of interleukin-6, TNF-${\alpha}$ were significantly decreased AR ethanol extract. iNOS, IL-$1{\beta}$, IL-6, TNF-${\alpha}$ mRNA expression of RAW 264.7 cells was increased depend on AR ethanol extract. Conclusions According to this study, AR ethanol extract has anti-oxidative and anti-inflammatoy effects.

Inhibitory Effects of Euphorbia supina Rafin on the Production of Pro-Inflammatory Mediator by LPS-Stimulated RAW 264.7 Macrophages (LPS로 활성화된 RAW 264.7 대식세포에서 애기땅빈대(Euphorbia supina Rafin)의 염증매개물질 억제효과)

  • Park, Sung-Chul;Son, Dae-Yeul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.4
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    • pp.486-492
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    • 2011
  • This study was designed to evaluate the effect of hot water extract (ESW) and 70% ethanol extract (ESE) from Euphorbia supina Rafin on LPS-stimulated inflammatory response in RAW 264.7 macrophages. Upon investigation at concentrations up to $1000\;{\mu}g$/mL, ESW and ESE did not have any cytotoxic effects on RAW 264.7 macrophages. ESW induced inhibition of 21.6%~54.8% of nitric oxide (NO) production at 100~1000${\mu}g$/mL, and $PGE_2$ production was inhibited up to 25.7%~38.2% at 250~1000${\mu}g$/mL, proportional to the ESW concentrations. ESW induced inhibition of 66.1% and 54.3% of IL-6 production at 250 and $1000\;{\mu}g$/mL, respectively. ESE (100~1000${\mu}g$/mL) induced inhibition of 38.3%~77.5% of NO, 40%~94.7% of $PGE_2$, and 43.9%~89.4% of IL-6 production, proportional to the ESE concentrations. Only 44.1% of IL-10 production was inhibited at a concentration of $500\;{\mu}g$/mL. ESE induced an increase in TNF-${\alpha}$ production at a concentration of 100 and $250\;{\mu}g$/mL, whereas at high concentrations (500 and $1000\;{\mu}g$/mL), ESE induced inhibition of 19.2% and 92.4% of TNF-${\alpha}$ production, respectively. In conclusion, concentrations of more than $500\;{\mu}g$/mL ESE demonstrated effective immune-modulating activity through inhibition of NO, $PGE_2$, IL-6, IL-10, or TNF-${\alpha}$ production, as it relates to the macrophage's immuno-activity; therefore, ESE has potential as a good candidate substance for reduction of inflammatory responses.

Effect of Artemisiae Argi Folium Fermented with Lactobacillus Pentosus and Saccharomyces Cerevisiae on TNF-${\alpha}$ Production in RAW 264.7 and HepG2 Cells (유산균 발효 애엽과 효모균발효 애엽 물추출물의 종양괴사인자-알파 생성촉진효과)

  • Kim, Youn-Sub;Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.6
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    • pp.956-961
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    • 2010
  • Tumor necrosis factor-alpha (TNF-${\alpha}$) is a major mediator of immuno-inflammatory activity. The purpose of this study is to investigate whether TNF-${\alpha}$ productions of mouse macrophage RAW 264.7 and human hepatocyte HepG2 are modulated by Artemisiae argi Folium water extract (AW), Lactobacillus pentosus-fermented Artemisiae argi Folium water extract (AFL), and Saccharomyces cerevisiae-fermented Artemisiae argi Folium water extract (AFS) for 3 h of incubation. Effect of AW on cell viability of HepG2 was also investigated. TNF-${\alpha}$ productions were measured by Enzyme-Linked Immnunosorbent Assay method and cell viability was measured by MTT assay. Both AFL and AFS significantly increased TNF-${\alpha}$ productions of RAW 264.7 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). Also, AFL and AFS significantly increased TNF-${\alpha}$ productions of HepG2 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). AW significantly increased TNF-${\alpha}$ production of HepG2 at the concentration of 100 and 200 ${\mu}g$/mL (p<0.05). AW did not show any cytotoxicity on HepG2 cells for 3 h. These results suggest that AFL, AFS, and AW have the immune-enhancing property related with its increasing effect on TNF-${\alpha}$ production of macrophage and hepatocyte.

Antioxidant and Anti-Inflammatory Effects of Kamisipjeondaebotang in RAW 264.7 Cells (가미십전대보탕의 RAW 264.7 세포에서 항산화 및 항염증 효과)

  • Myung, Jeong-Ho;Lee, Myung-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.11
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    • pp.1271-1277
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    • 2017
  • As Kamisipjeondaebotang (KSD) extract is an herbal ingredient, safety is very important due to possible cell poisoning or heavy metal toxicity to organs when administered to humans or animals. Accordingly, this study examined the antioxidant and anti-inflammatory effects of KSD extract to confirm its medicinal safety by using RAW 264.7 cells after heavy metal screening, functional index test of the liver and kidney, and cell survival rate test. Heavy metals were not found in KSD extracts or were less than standard amounts. Liver function indices such as aspartate aminotransferase and alanine aminotransferase revealed low values and kidney function indices such as creatinine and blood urea nitrogen were not significantly different from the normal group. This proved the safety to the human. RAW 264.7 cells showed no poisoning compared to the control group in terms of survival rate. Regarding the antioxidant effect of KSD extract, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazo-line-6-sulphonic acid) radical scavenging activity increased at concentrations over $10{\mu}g/mL$. The anti-inflammatory effect of KSD extract significantly decreased based on the amount of nitric oxide at concentrations of 10 and $100{\mu}g/mL$ compared to the control group. Expression of interleukin (IL)-$1{\beta}$ and IL-6 decreased in a concentration-dependent manner. There was no significant difference in tumor necrosis factor-${\alpha}$ level. Based on the results, KSD can be regarded as a safe antioxidant with anti-inflammatory effects for fracture treatment.

Anti-inflammatory and Antioxidant Effects of Water Extracts of Sasangja-tang(SSJ) and Gami-sasangja-tang(GSJ) (사상자탕과 가미사상자탕의 항염증 및 항산화 효과 비교 연구)

  • Choi, Jung-Eun;Park, Bo-Kyung;Jin, Mirim
    • Journal of Haehwa Medicine
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    • v.23 no.2
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    • pp.5-13
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    • 2015
  • Objectives : The anti-inflammatory and antioxidant effects of water extracts of Sasangja-tang(SSJ) and Gami-sasangja-tang(GSJ) were investigated. The effects of SSJ and GSJ were compared. Methods : We performed cell viability assay in HaCaT cells and RAW 264.7 cells using 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide(MTT) assay. We measured chemokines(regulated on activation normal T-cell expression and secreted ; RANTES/CCL5, interferon-inducible protein; IP-10/CXCL10, macrophage-derived chemokine; MDC/CCL22) in HaCat cells, also we measured cytokines (tumor necrosis factor-${\alpha}$; TNF-${\alpha}$, interleukin-6; IL-6) and nitric oxide(NO) production in RAW 264.7 cells using enzyme-linked immunosorbent assay(ELISA) and NO assay. Western blot assay was used to evaluate the expression for inducible nitric oxide synthase(iNOS) in RAW 264.7 cells. Results : SSJ and GSJ did not affect the cell viability at the concentrations treated ($0-800{\mu}g/ml$). As a result of SSJ and GSJ treatment in HaCat cells stimulated by TNF-${\alpha}$(10 ng/ml) and interferon(IFN)-${\gamma}$(10 ng/ml), the production of RANTES and IP-10 was inhibited significantly. However there was no significant difference in the secretion of MDC. And in RAW 264. 7 cells stimulated by lipopolysaccharide(LPS, $1{\mu}g/ml$), SSJ and GSJ treatment significantly inhibited the secretion of TNF-${\alpha}$ and IL-6 and the production of NO. The expression of iNOS was also decresed by SSJ and GSJ treatment in RAW 264. 7 cells. Compared with SSJ, GSJ was superior to SSJ in inhibition of RANTES, IP-10, TNF-${\alpha}$, IL-6 and NO production at the concentration of $200{\mu}g/ml$. Conclusion : Both SSJ and GSJ have anti-inflammatory and antioxidant effects. And GSJ has better effects than SSJ.

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