• The Journal of Korean Obstetrics and Gynecology
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• v.34 no.3
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• pp.15-28
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• 2021

Objectives: This study was designed to examine the anti-cancer activity by innate immunomodulating and anti-inflammatory effects of liriopis tuber water extract (LPE). Methods: Cell cytotoxicity was tested with 4T1 mouse mammary carcinoma cells, spleen cells, macrophage, and RAW264.7 cells. To investigate innate immunomodulating effects of LPE on macrophage, we measured tumor necrosis factor-alpha (TNF-α), interleukin-12 (IL-12), and interleukin-10 (IL-10). To investigate innate immunomodulating effects of LPE on RAW264.7 cell, we measured TNF-α, interleukin-6 (IL-6). In addition, TNF-α and nitric oxide (NO) induced by lipopolysaccharide (LPS) were measured after treating with LPE to observe innate immunomodulating effect of LPE on RAW264.7 cell. Also, mitogen-activated protein kinase (MAPK) and nuclear factor κB (NF-κB) were examined by western blot analysis. Results: In an in vitro cytotoxicity analysis, LPE affected tumor cell growth above specific concentration. As compared with the control group, the production of TNF-α, IL-12, and IL-10 were increased in macrophage. As compared with the control group, the production of TNF-α and IL-6 were increased in RAW 264.7 cell. The expression of TNF-α and NO induced by LPS after treating LPE was decreased. In addition, treatment of RAW 264.7 cell with LPE increased the phosphorylation levels of p-extracellular signal-regulated kinase (p-ERK), p-Jun N-terminal kinase (p-JNK), and p-p38. Conclusions: LPE might have impact on the anti-cancer effect by activation of innate immune system and inflammation control.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.2
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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.

• 한국약용작물학회:학술대회논문집
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• 2008.11a
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• pp.354-355
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• 2008

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1671-1676
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• 2012

In this study, the immuno-enhancing activity of seed extracts were studied on the macrophage cell lines. We examined the effect of nine seed extracts on nitric oxide (NO) production in RAW 264.7 cells and selected four highly-effective seed candidates (Fagopyrum esculentum, Taraxacum platycarpum, Impatiens balsamina, Helianthus annuus) for further immune-related studies. The effects of the four seed extracts on the production of immune-related cytokines in the RAW 264.7 macrophage cell line and proliferation of Molt-4 as a T cell line were investigated. The secretion of NO from the RAW 264.7 cells was increased up to 39 ${\mu}M$ by adding the seed extracts (25 ${\mu}g/mL$) compared to the control. Also, the secretion of tumor necrosis factor-alpha (TNF-${\alpha}$) was also increased up to 32 times by adding the seed extracts (25 ${\mu}g/mL$). Secretion of cytokines such as interleukin-1 beta (IL-$1{\beta}$), interleukin-6 (IL-6), and interleukin-10 (IL-10) was also increased and induced the proliferation of T cells compared to the control. In conclusion, these results suggest that four seed extracts provide beneficial immuno-enhancing effects for human health.

• The Korean Journal of Oral and Maxillofacial Pathology
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• v.41 no.3
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• pp.121-129
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• 2017

Coffee (Coffea spp.) is one of the most important agricultural commodities, being widely consumed in the world. Various beneficial health effects of coffee have been extensively investigated, but data on habitual coffee consumption and its bio-physiological effect have not been clearly explained as well as it is not proved the cause and effect between drinking coffee and its bio-physiological reactions. We made the dialyzed coffee extract (DCE), which is absorbable through gastrointestinal tract, in order to elucidate the cellular effect of whole small coffee molecules. RAW 264.7 cells, a murine macrophage lineage, were directly treated with DCE, i.e., DCE-2.5 (equivalent to 2.5 cups of coffee a day), DCE-5, and DCE-10, for 12 hours, and their protein extracts were examined by immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the inflammation-related proteins depending on the doses of DCE. RAW 264.7 cells treated with DCE showed marked increase of cathepsin C, cathepsin G, CD20, CD28, CD31, CD68, indicating the activation of innate immunity. Particularly, the macrophage biomarkers, cathepsin G, cathepsin C, CD31, and CD68 were markedly increased after DCE-5 and DCE-10 treatments, and the lymphocyte biomarkers, CD20 and CD28 were consistently increased and became marked after DCE-10 treatment. On the other hand, RAW 264.7 cells treated with DCE showed consistent increase of IL-10, an anti-inflammatory factor, but gradual decreases of different pro-inflammatory proteins including $TNF{\alpha}$, COX-2, lysozyme, MMP-2, and MMP-3. In particular, the cellular signaling of inflammation was gradually mitigated by the reduction of $TNF{\alpha}$, COX-2, IL-12, and M-CSF, and also the matrix inflammatory reaction was reduced by marked deceases of MMP-2, MMP-3, and lysozyme. These anti-inflammatory expressions were consistently found until DCE-10 treatment. Therefore, it is presumed that DCE may have dynamic effects of innate immunity activation and pro-inflammation suppression on RAW264.7 cells simultaneously. These effects were consistently found in the highest dose of coffee, DCE-10 (equivalent to 10 cups of coffee a day in man), that might imply the small coffee molecules were accumulated in RAW 264.7 cells after DCE-10 treatment and produce synergistic cytokine effects for innate immunity activation and anti-inflammatory reaction concurrently.

• The Korea Journal of Herbology
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• v.26 no.1
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• pp.53-58
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• 2011

Objectives : The purpose of this study is to investigate effects of Scutellariae Radix Water Extract on hydrogen peroxide production in RAW 264.7 mouse macrophages. Methods : Scutellariae Radix produced from South Korea (SK) and Scutellariae Radix produced from China (SC) were extracted by hot water. Effects of SK and SC on hydrogen peroxide production in RAW 264.7 were measured by dihydrorhodamine 123 assay after 2, 4, 20, 24, 28, 44, and 48 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL. Results : SK significantly increase hydrogen peroxide production in RAW 264.7 cells for 2, 4, 20, 24, 28, 44, and 48 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL (P < 0.05). SC also significantly increase hydrogen peroxide production in RAW 264.7 cells for 4, 20, 24, 28, and 48 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL (P < 0.05). For 2 h incubation, SC significantly increase hydrogen peroxide production in RAW 264.7 cells at the concentrations of 10, 25, and 100 ug/mL (P < 0.05). For 44 h incubation, SC significantly increase hydrogen peroxide production in RAW 264.7 cells at the concentrations of 10, 25, and 50 ug/mL (P < 0.05). Conclusions : These results suggest that Scutellariae Radix has the immune - enhancing property related with its increasement of hydrogen peroxide production in macrophages.

• Korean Journal of Acupuncture
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• v.26 no.3
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• pp.69-76
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• 2009

Objectives : 본 연구의 목적은 제습, 해독, 통리관절등의 효능이 있는 토복령이 RAW264.7 대식세포주에서 lipopolysaccharide(LPS)로 처치하여 생성되는 nitric oxide(NO)와 prostaglandin $E_2$($PGE_2$)에 억제작용이 있는지 관찰하고자 하는 것이다. Methods : 토복령이 RAW264.7 대식세포주에 세포독성이 유무를 관찰하기 위하여 농도별 MTT assay를 수행하여 세포생존율을 측정하였다. 또한 LPS로 염증유발된 RAW264.7 대식세포주에서 토복령의 농도별 처치가 NO 및 $PGE_2$ 생성억제에 미치는 영향을 관찰하고자 NO 및 $PGE_2$ assay를 수행하였다. Results : 토복령의 농도별 처치가 RAW264.7 대식세포주에서 세포독성을 유발하는지 MTT assay로 측정한 결과 세포독성은 관찰되지 않았으며, 토복령은 LPS처치로 인하여 증가된 NO 및 $PGE_2$ 생성을 통계학적으로 유의하게 감소시킴을 관찰하였다. Conclusions : 본 연구를 통하여 토복령은 RAW264.7 대식세포주에서 LPS로 유도된NO 및 $PGE_2$ 생성을 효과적으로 억제시킴으로써 추후 염증질환의 치료제로서 가능성을 확인하였다.

• The Journal of Korean Medicine
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• v.30 no.6
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• pp.69-79
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• 2009

Objective: In this study, the immunomodulatory activity of a mixture of wild Panax ginseng and red-mold rice extracts (MPR) on RAW 264.7 macrophage cells in the presence and absence of methotrexate (MTX), an anti-cancer drug, was investigated. Methods and Results: In the cell viability, MPR showed a significant cell proliferation and inhibited cell regression by red-mold rice (RMR) alone or MTX alone. MPR induced moderate increase in nitric oxide (NO) production. NO production and inducible nitric oxide synthase (iNOS) mRNA expression by LPS decreased after MPR treatment. In addition, MPR slightly induced COX-2 mRNA expression, but it did not affect the expression of COX-2 mRNA by LPS treatment. In RT-PCR analyses, MPR induced IL-$1{\alpha}$, IL-$1{\beta}$, IL-6, and TNF-$\alpha$ mRNA expression, but had no effect on IL-10 and TGF-$\beta$, regardless of MTX treatment. Furthermore, MPR did not interfere with the cytotoxicity of MTX against MCF-7 human breast carcinoma cells. Conclusions: MPR is efficacious in protecting against MTX-induced cell regression as a result of macrophage activation, resulting in induction of cytokine expression, implying that MPR could be considered an adjuvant in MTX-chemotherapy.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.947-954
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• 2005

This study was conducted to investigate the effects of hydrolyzed Lactobacillus supplementation with digestive enzymes treatment on the macrophage activation, the induction of nitric oxide(NO), interleukin (IL)-6 and tumor necrosis factor(TNF)-$\alpha$. The RAW 264.7 murine macrophage was exposed to porcine Lactobacillus strains which were digested with both pepsin and pancreatin. The production of NO, TNF-$\alpha$ and IL-6 in the macrophage were strain and dose-dependent, respectively. The induction of NO and cytokines were higher in both 3149 and 3146 strains compared with other Lactobacillus strains. Overall, the level of NO was observed at the lower range between 10 and 150 μg hydrolysates per ml, whereas IL-6 and TNF-$\alpha$ were observed at relatively higher concentration between 50 and 300 μg hydrolysates per ml. Lactobacillus strains which produced a high level of NO also showed a high induction of TNF-$\alpha$ and IL-6. Therefore, the present results suggest that hydrolysates of Lactobacillus strains are related to induction of several macrophage mediators, and then it could be beneficially used to modulate gastrointestinal immune function of the host. Also, the methodogly employed in this study might be useful to investigate the effects of lactic acid bacteria on gastrointestinal immunity.

• The Korea Journal of Herbology
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• v.21 no.2
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• pp.165-173
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• 2006

Objectives : Coptidis Rhizoma has been known traditional medicine with antimicrobial activities. We investigated inhibitory effects of Coptidis Rhizoma extract on lipopolysaccharide(LPS)-induced nitric oxide production from mouse macrophages. Methods : After Coptidis Rhizoma extract was pretreated in BV2, mouse brain macrophages and RAW264.7 mouse macrophages, cells were activated with LPS. To investigate cytotoxicity Coptidis Rhizoma extract, cell viability was measured by MTT assay. The production of nitric oxide(NO) and inducible nitric oxide synthase(iNOS) was determined in each culture supernatant and mRNA by Griess reaction and RT-PCR. The production of $TNF-{\alpha}$ from cells was measured by ELISA. Results : Coptidis Rhizoma extract significantly inhibited LPS-induced NO production in BV2 and RAW264.7 cells. Coptidis Rhizoma extract also greatly suppressed mRNA expression of iNOS in BV2 and RAW264.7 cells activated by LPS. Conclusion : These data suggests that Coptidis Rhizoma extract may have an anti-inflammatory effect through the inhibition of NO production.