• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.1
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• pp.108-119
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• 2009

Objectives : This study was carried out to investigate the effects of Samhwangseje(SHSJ) on anti-Inflammation in Raw 264.7 cell. Methods : The effects of SHSJ on anti-Inflammation were measured by the cytotoxicity of Raw 264.7 cell, the inhibition for NO, TNF-$\alpha$, $PGE_{2}$, iNOS and COX-2, the blocking NF-${\kappa}B$ into nucleus. Results : 1. All concentrations of SHSJ had no cytotoxicity in Raw 264.7 cell. 2. All concentrations of SHSJ inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of SHSJ did not inhibit the production of TNF-$\alpha$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of SHSJ inhibited the production of $PGE_{2}$ in the Raw 264.7 cell stimulated with LPS. 5. All concentrations of SHSJ did not inhibit the expression of COX-2 but concentrations of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$ SHSJ inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. Concentrations of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$ SHSJ had the effect of blocking NF-${\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.3
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• pp.1-15
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• 2012

Objectives: The purpose of this study was to investigate the effects of Chaenomelis Fructus Herba Water Extract(CF) on the production of inflammatory mediators in RAW 264.7 cell mouse macrophages stimulated with LPS. Methods: We have not examined effect of CF on the cell viability of RAW 264.7 cell until we investigated effects of CF on LPS-induced productions of NO, Ca and various cytokines in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically(P<0.05). Results: 1. CF increased the cell viability in the RAW 264.7 cell at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 2. CF inhibited significantly increasing the production of NO in LPS-induced RAW 264.7 cell at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 3. CF inhibited significantly increasing the production of Intracellular Ca in LPS-induced RAW 264.7 cell at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 4. CF inhibited significantly the IL-2, IL-10, IL-12p70, TNF-${\alpha}$, GM-CSF, M-CSF, LIF and VEGF of the RAW 264.7 cell induced by LPS at the density of 25, 50, 100 and 200 ${\mu}g/ml$. 5. CF inhibited significantly the IL-4 at the density of 25, 50 ${\mu}g/ml$, the IL-5, IL-15 and MIG at the density of 25, 50 and 200 ${\mu}g/ml$ and IFN-${\gamma}$ at the density of 25, 100 ${\mu}g/ml$ respectively in the RAW 264.7 cell increased by LPS. Conclusions: CF inhibited significantly increasing IL-2, IL-10, IL-12p70, TNF-${\alpha}$, GM-CSF, M-CSF, LIF, VEGF, NO and Ca in LPS-induced RAW 264.7 cell at the density of more than 25 ${\mu}g/ml$ without causing the toxicity. These results signify that CF has antiinflammatory effect on controlling the over inflammatory reaction by the RAW 264.7 cell.

• The Korea Journal of Herbology
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• v.28 no.6
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• pp.129-133
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• 2013

Objectives : The purpose of this study was to investigate the effects of Angelicae acutilobae Radix Water Extract (AA) on the production of cytokines in RAW 264.7 cell stimulated with lipopolysaccharide (LPS). Method : RAW 264.7 cells were cotreated with AA (50 and $100{\mu}g/mL$) and lipopolysaccharide (LPS; $1{\mu}g/mL$) for 24 hours. After 24 hours treatment, using bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, tumor necrosis factor-alpha(TNF-${\alpha}$) granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), and macrophage inflammatory protein(MIP)-$1{\alpha}$ were measured. Result : AA significantly inhibited LPS-induced production of IL-6 and MIP-$1{\alpha}$ from LPS-stimulated RAW 264.7 cells at the concentration of $50{\mu}g/mL$. AA significantly inhibited LPS-induced production of TNF-${\alpha}$ from LPS-stimulated RAW 264.7 cells at the concentration of $100{\mu}g/mL$. AA significantly inhibited LPS-induced production of G-CSF and GM-CSF in RAW 264.7 cells at the concentrations of 50 and $100{\mu}g/mL$. Conclusion : These results suggest that AA has anti-inflammatory effect related with its inhibition of proinflammatory cytokines such as IL-6, TNF-${\alpha}$, G-CSF, GM-CSF, and MIP-$1{\alpha}$ in LPS-induced macrophages.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.3
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• pp.330-336
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• 2014

This study aimed at examining the anti-inflammatory effects of Scutellariae Radix & Lonicerae Caulis water extract(SC). RAW 264.7 mouse macrophage cells were treated with $25{\sim}200{\mu}g/m{\ell}$ SC for 24 hours. Cell viability was then measured using MTT assays. The nitric oxide(NO) production and the creation of several cytokines in LPS-stimulated RAW 264.7 cells were investigated. SC inhibited significantly increasing the production of NO in LPS-induced RAW 264.7 cell at the density of 25, 50 and $200{\mu}g/m{\ell}$. SC inhibited significantly the TNF-${\alpha}$ of the RAW 264.7 cell induced by LPS at the density of $50{\mu}g/m{\ell}$. SC inhibited significantly the MIP-$1{\alpha}$ of the RAW 264.7 cell induced by LPS at the density of 25, 50 and $100{\mu}g/m{\ell}$. SC inhibited significantly the MIP-$1{\beta}$, MIP-2 at the density of 50, $100{\mu}g/m{\ell}$ in the RAW 264.7 cell increased by LPS, respectively. SC did not affect the production levels of VEGF in RAW 264.7 cell. As a result, SC significantly inhibited the inductions of MIP-$1{\alpha}$, MIP-$1{\beta}$, MIP-2 and NO in LPS-induced RAW 264.7 cell without causing the toxicity. These results signify that SC has anti-inflammatory effects on controlling the over inflammatory reaction on the RAW 264.7 cell.

• Journal of Life Science
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• v.28 no.9
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• pp.1022-1029
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• 2018

This study was carried out to search for the anti-inflammatory activities of ethanol extracts obtained from 5 kinds of oriental medical plant; Pleuropterus multiflorus extract (PME), Acorus calamus L. extract (ACE), Lithospermum erythrorhizon Siebold & Zucc. extract (LEE), Xanthium strumarium L. extract (XSE), Lonicera japonica extract (LJE), which have traditionally been used as a drug in oriental medical plants in Korea. XSE showed cytotoxicity at 100, $200{\mu}g/ml$ concentration in RAW264.7 cells (p<0.05) and ACE showed cytotoxicity at $200{\mu}g/ml$ concentration in RAW264.7 cells (p<0.05). But other oriental medical plants did not showed cytotoxicity was observed in RAW264.7 cells below $200{\mu}g/ml$ concentration. These extracts at non-toxic concentrations showed anti-inflammatory effects. PME, ACE, XSE and LJE showed a concentration-dependent inhibitory effect on NO production and $PGE_2$ production in LPS-induced RAW264.7 cells. In particular, XSE showed the highest NO production inhibition ($52.9{\mu}g/ml$, $IC_{50}$) as well as the highest $PGE_2$ production inhibition at $50{\mu}g/ml$ (73.6%). ACE and LEE showed cell proliferation effects on HaCaT keratinocyte cells. Especially, LEE showed 21.1, 53.5 and 99.6% proliferative activity by incubation for 1, 3, 5days at $100{\mu}g/ml$ concentration. ACE also showed 11.2, 26.0% proliferative activity for 1day and 3days at $10{\mu}g/ml$ concentration. As a result of this study, ethanol extracts obtained from 5 kinds of oriental medical plant showed anti-inflammatory activity and HaCaT cell regeneration effect.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.19-26
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• 2017

Effects of resveratrol glucosylation on the immunomodulation properties of resveratrol and on the viability of macrophage cells have been studied by using murine macrophage RAW 264.7 cells. Nitric oxide (NO) and interleukin 6 (IL-6) expression in macrophages in vitro were studied after treatment with different concentrations of (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), or (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G). In vitro viability of RAW 264.7 cells after treatment with the aforementioned three compounds was also studied. As demonstrated by macrophage cell viability assays, two different resveratrol monoglucosides, R-3-G and R-4'-G, exhibited 50-80% reduced cytotoxicity in comparison to (E)-resveratrol in A549 and HepG2 cells. Compared to the resveratrol aglycon, both glucosylated resveratrol derivatives positively modulated NO and IL-6 production in macrophages positively via transcriptionally up-regulating IL-6 and iNOS expression. Conjugation of a glucose moiety on resveratrol was found to enhance the immunomodulating activity of resveratrol and the viability of RAW 264.7 cells.

• Journal of Acupuncture Research
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• v.36 no.4
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• pp.256-263
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• 2019

Background: The purpose of this study was to investigate the anti-inflammatory response of lipopolysaccharide (LPS) activated macrophages (RAW 264.7 murine cell line) to JCE003 which is an extract including Eucommia ulmoides, Juglans regia, Eleutherococcus senticosus, and Zingiber officinale. Methods: An MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide] assay was performed to analyze the survival rate of RAW 264.7 cells. The production of nitric oxide and pro-inflammatory cytokines (IFN-${\gamma}$, TNF-${\alpha}$, IL-$1{\beta}$, IL-6) in LPS-induced RAW 264.7 cells was measured by enzyme-linked immunosorbent assay. mRNA expression levels of pro-inflammatory cytokines (IFN-${\gamma}$, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6) were analyzed by quantitative polymerase chain reaction analysis. Results: Exposure of LPS-activated RAW 264.7 cells to JCE003 was not cytotoxic up to $400{\mu}g/mL$, but cell survival was statistically significantly decreased at $800{\mu}g/mL$ (p < 0.001). Nitric oxide production was not markedly lowered in LPS-activated RAW 264.7 cells by exposure to JCE003 (10, 50, 100, 200, 400, $800{\mu}l/mL$) compared with the Control group. In addition, JCE003 reduced the production of TNF-${\alpha}$ in LPS-induced RAW 264.7 cells at $400{\mu}g/mL$ (p < 0.05), but IFN-${\gamma}$ and TNF-${\alpha}$ mRNA expression in LPS-induced RAW 264.7 cells was decreased at 100, 200, and $400{\mu}g/mL$ JCE003 (p < 0.01). Conclusion: These results suggest that JCE003 inhibited the expression and production of pro-inflammatory cytokines in LPS-activated RAW 264.7 cells. The findings of this study provide basic data for the development of new Korean medicine anti-inflammatory drugs.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.2
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• pp.12-22
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• 2012

Objectives: The aim of this study is to investigate immune enhancing effect of Houttuyniae Herba water extract(HW) on RAW 264.7 cell of mouse macrophages. Methods: Effects of HW on productions of nitric oxide(NO) and hydrogen peroxide($H_2O_2$) in RAW 264.7 mouse macrophages were measured. Effect of HW on production of cytokines such as interleukin(IL)-$1{\beta}$, IL-6, and tumor necrosis factor(TNF)-${\alpha}$ in RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. All of results were represented P<0.05 compared to the normal. Results: 1. After 24 hr incubation, HW increased significantly NO production in RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ${\mu}g$/mL. 2. After 24 hr incubation, HW increased significantly hydrogen peroxide production in RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ${\mu}g$/mL. 3. After 24 hr incubation, HW increased significantly IL-$1{\beta}$ production in RAW 264.7 cells at the concentrations of 100 and 200 ${\mu}g$/mL. 4. After 24 hr incubation, HW increased significantly IL-6 production in RAW 264.7 cells at the concentrations of 100 and 200 ${\mu}g$/mL. 5. After 24 hr incubation, HW increased significantly TNF-${\alpha}$ production in RAW 264.7 cells at the concentrations of 50, 100, and 200 ${\mu}g$/mL. Conclusions: These results suggest that HW has immune enhancing activity related with its increasement of NO, hydrogen peroxide, IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ in macrophages.

• The Korea Journal of Herbology
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• v.32 no.3
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• pp.97-103
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• 2017

Objective : According to recent studies, Anemarrhenae Rhizoma has anti-inflammatory activities of DW extract, but it hasn't not yet conducted to evaluate inflammatory factors about 80% ethanol extract. Therefore, The aim of this study is to investigate the various effects of individual or combined 80% ethanol extract of Anemarrhenae Rhizoma on cell viability and various anti-inflammatory factors. Methods : Anemarrhenae Rhizoma extract was prepared with 80% ethanol. MTT assay, ELISA, and Luminex were performed in LPS-activated RAW 264.7 cell line to measure cytotoxicity, Nitric oxide (NO), cyclooxygenase-2 (COX-2), prostaglandin E2 ($PGE_2$), Leukotriene B4 ($LTB_4$), and cytokines ($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$), respectively. Results : At concentration of $200{\mu}g/m{\ell}$ Anemarrhenae Rhizoma extract, cytotoxicity was observed in RAW 264.7 cells. However, at concentration less than $100{\mu}g/m{\ell}$ of Anemarrhenae Rhizoma, cytotoxicity was not observed in RAW 264.7 cells. All concentration of Anemarrhenae Rhizoma extract showed no difference of NO, and $IL-1{\beta}$level in RAW 264.7 cells compared with control group. In contrast, at concentration of $100{\mu}g/m{\ell}$ Anemarrhenae Rhizoma extract significantly inhibited LPS-induced production of COX-2, PGE2, and $LTB_4$ level in RAW 264.7 cells. In addition, the production of proinflammatory cytokines (IL-6, $TNF-{\alpha}$) in LPS-induced RAW 264.7 cells was significantly decreased at concentration of all or 10, and $100{\mu}g/m{\ell}$, respectively. Conclusion : These findings demonstrate that Anemarrhenae Rhizoma has inhibitory effect on inflammatory mediators in LPS-activated RAW 264.7 cells showing possible developed as a raw material for new therapeutics to ease the symptoms related with inflammatory.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.2 s.33
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• pp.77-88
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• 2007

Objectives : This experimental study was performed to investigate the effects of Yeouigeumhwang-san(YUGHS) on anti-inflammation and anti-Propionibacterium acnes. Methods : The cytotoxicity of YUGHS about viability of Raw 264.7 cell was tested by using a colorimetric tetrazolium assay(MTT assay). To investigate the anti-inflammatory effets of YUGHS on LPS-induced macrophage Raw 264.7 cell, we used ELISA kit and Western blots. Inhibitory effects of YUGHS on Propionibactrium acnes were investigated by using paper disk diffusion method. Results : 1. YUGHS has no cytotoxicity under 50 ${\mu}g/ml$ concentration but over 50 ${\mu}g/ml$ has a little cytotoxicity in Raw 264.7 cell. 2. Concentration of 100 ${\mu}g/ml$ YUGHS inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of YUGHS did not inhibit the production of $TNF-{\alpha}$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of YUGHS significantly inhibited the production of $PGE_2$ in the Raw 264.7 cell stimulated with LPS. 5. YUGHS did not inhibit the expression of COX-2 but concentration of 50 ${\mu}g/ml$ YUGHS inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. YUGHS has the effect of blocking $NF-{\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell 7. YUGHS did not have the inhibitory effect of Propionibactrium acnes. Conclusions : These results indicate that Yeouigeumhwang-san has anti-inflammatory effets. If further study is performed, the use of Yeouigeumhwang-san will be valuable and benificial in the therapy of acnes.