• 한방안이비인후피부과학회지
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• 제22권1호
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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.

• 대한한방부인과학회지
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• 제25권3호
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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.

• 대한본초학회지
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• 제28권6호
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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.

• 동의생리병리학회지
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• 제28권3호
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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.

• 생명과학회지
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• 제28권9호
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• pp.1022-1029
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• 2018

이 연구는 한국에서 전통적으로 사용되어 온 5종의 한약재인 하수오(Pleuropterus multiflorus), 창포(Acorus calamus L.), 지치(Lithospermum erythrorhizon Siebold & Zucc.), 창이자(Xanthium strumarium L.), 인동초(Lonicera japonica) 에탄올 추출물의 항염증에 미치는 효과를 탐색하였다. 창이자는 RAW264.7 세포에 100, $200{\mu}g/ml$ 농도에서 세포 독성을 나타냈고(p<0.05), 창포는 RAW264.7 세포에 $200{\mu}g/ml$ 농도에서 세포 독성을 나타냈다(p<0.05). 다른 한약재들은 RAW264.7 세포에 $200{\mu}g/ml$ 농도 이하에서 세포 독성이 나타나지 않았다. 독성이 없는 농도에서 5종의 한약재 추출물들의 항염증 효과를 확인하였다. 하수오, 창포, 창이자, 인동초는 LPS 유도된 RAW264.7세포에서 NO 생산과 $PGE_2$생산에 대해 농도 의존적으로 억제 효과를 보였다. 특히, 창이자는 $52.9{\mu}g/ml$ ($IC_{50}$)로 가장 뛰어난 NO 생성 저해효과를 나타냈을 뿐만 아니라 $50{\mu}g/ml$ 농도 구간에서 가장 뛰어난 $PGE_2$ 생성 저해능을 나타냈다(73.6%). 창포와 지치는 HaCaT 세포에 대하여 세포가 증식하는 효과를 나타냈다. 특히 지치는 $100{\mu}g/ml$ 농도구간에서 1, 3, 5일 배양 시 21.1, 53.5, 99.6%의 증식능을 보였다. 창포 또한 $10{\mu}g/ml$ 농도구간에서 1, 3일 배양 시 11.2, 26.0%의 증식능을 보였다. 따라서 본 연구를 통해 5종의 한약재 에탄올 추출물의 항염증 활성 및 HaCaT 세포 재생에 미치는 효과를 확인하였다.

• 한국미생물·생명공학회지
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• 제45권1호
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• pp.19-26
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• 2017

레스베라트롤의 면역제어 성질과 대식세포의 생존능력과 관련하여 당화된 레스베라트롤의 효과를 확인하기 위해 대식세포 RAW 264.7에서 연구하였다. 인비토로에서 대식세포에서 총 4개의 레스베라트롤 및 당화된 유도체 (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), 및 (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G)를 여러 가지 농도로 처리한 후 일산화질소 (NO)와 인터루킨 6 (IL-6) 발현을 연구하였다. 앞서 언급한 물질로 처리한 후 인비토로에서 RAW 264.7 세포의 생존능력도 연구하였다. 대식세포 생존능력 평가분석 결과를 보면, 두 개의 레스베라트롤 모노글루코사이드인 R-3-G와 R-4'-G은 (E)-resveratrol와 비교하여 A549 and HepG2 세포에서 50-80% 감소된 독성을 보여준다. 당이 없는 레스베라트롤과 비교하면, 당화된 레스베라트 유도체는 긍정적으로 전사적으로 IL-6 및 iNOS 발현이 높아지는 방향으로 NO 및 대식세포에서 IL-6의 생산이 조절된다. 레스베라트롤의 당의 역할은 RAW 264.7 세포의 생존능력과 레스베라트롤의 면역제어 활성을 증가시켜 주는 것으로 보여주고 있다.

• Journal of Acupuncture Research
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• 제36권4호
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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.

• 대한한방부인과학회지
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• 제25권2호
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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.

• 대한본초학회지
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• 제32권3호
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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.

• 한방안이비인후피부과학회지
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• 제20권2호통권33호
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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.