• The Korea Journal of Herbology
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• v.28 no.6
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• pp.111-117
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• 2013

Objectives : This research aimed at studying the immuno modulating activity of Fermented Epimedii Herba (EHS). Method : The impacts on the cell viability, hydrogen peroxide and nitric oxide (NO) generation in cells, and cytokines such as tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin (IL)-6, IL-$1{\beta}$, monocyte chemoattractant protein-1 (MCP-1) level have been measured by using Raw 264.7 cells with the specimen EHS as the fermented extract of Epimedii Herba with Saccharomyces cerevisiae STV89. Result : As a result of MTT assay to confirm the cytotoxicity of extracts from fermented Epimedii Herba, the toxicity was not excessively induced in Raw 264.7 cells when EHS were processed by concentration. EHS increased hydrogen peroxide generation in Raw 264.7 cells. EHS suppressed NO generation in Raw 264.7 cells while they significantly suppressed the increase of NO generation induced by LPS in macrophage. EHS significantly decreased the generation amount of TNF-${\alpha}$ and IL-6 induced by LPS in Raw 264.7 cells at $25{\mu}g/mL$ or more. Conclusion : It appeared that the fermented extract of Epimedii Herba manufactured from Epimedii Herba significantly has the immuno modulating acitivity as it did not excessively trigger cytotoxicity to Raw 264.7 cells, increased hydrogen peroxide generation in Raw 264.7 cells, decreased NO generation in macrophage, and especially, suppressed both TNF-${\alpha}$ and IL-6 generation in macrophage induced by LPS.

• YAKHAK HOEJI
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• v.49 no.6
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• pp.459-464
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• 2005

The immune reinforcement of the probiotic lactic acid bacteria Lactobacillus acidophilus, Streptococcus thermophilus and Bifidobacterium bifidum was studied in RAW 264.7 cell line treated with diluted solution (dilution to $2^{5}$) of the supernatnats of lactic acid bacteria. RAW 264.7 cell line was used as a macrophage model to assess the effects of lactic acid bacteria on the production of nitric oxide (NO) and cytokine tumor necrosis factor (TNF)-$\alpha$ and cell morphological changes. The production of NO and TNF-$\alpha$ were largely affected by lactic acid bacteria in dose-dependent manner in 24 or 48 hr cultures and cell morphological changes were also largely affected by lactic acid bacteria. Especially Bifidobacterium bifidum differentially stimulated the production of NO and TNF-$\alpha$. NO production was increased by Bifidobacterium bifidum 25 $\mu$l/ml more than LPS (20 ng/ml) control, and TW-$\alpha$ by Bifidobacterium bifidum 6.25 $\mu$l/ml more than LPS (10 ng/ml) control. The in vitro approaches employed here should be useful in further characterization of the effects of lactic acid bacteria on systemic immunity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.11
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• pp.1521-1527
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• 2012

Medicinal herbs have long been used as a remedy for diverse diseases in Asia owing to their various pharmacological effect. In this study, the immuno-enhancing activity of medicinal herbs was investigated using macrophage cell lines. Specifically, we examined the effects of extracts of twelve medicinal herbs on nitric oxide (NO) production in RAW 264.7 cells, and selected five that were highly effective (Glycyrrhiza glabra, Rehmannia glutinosa, Angelica gigas, Platycodon grandflorum, and Actinidia polygama) for further immune related studies. The effects of extracts from five theses medicinal herbs, which were mainly composed of polysaccharides and proteins on the production of immune-related cytokines in the RAW 264.7 macrophage cell line and the Molt-4 T cell line were investigated. The extracts of all investigated medicinal herbs increased the production of NO and cytokines such as tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-1beta (IL-$1{\beta}$), interleukin-6 (IL-6) and interleukin-10 (IL-10). Additionally, they slightly increased the proliferation of T-cells when compared to the control. Overall, the result of this study suggests that the five medicinal herb extracts investigated herein are useful natural immune enhancing agents.

• Archives of Pharmacal Research
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• v.25 no.6
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• pp.895-902
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• 2002

Monocytes and macrophages playa major role in defense mechanism of the host response to tumor, in part through the secretion of several potent products and macrophage cytokines. Monocytes and tissue macro phages produce at least two groups of protein mediators of inflammation, interleukin 1 (IL-1) and tumor necrosis factor (TNF). Recent studies emphasizes that TNF and IL-1 modulate the inflammatory function of endothelial cells, leukocytes, and fibroblasts. In this study, our work is directed toward studying the in vitro effects of Korean propolis on the ability to induce cellular and secretory responses in murine macrophage cell line, RAW 264.7. It was found that Water Extract of Korean Propolis (WEP) could activate macro phages by producing cytokines. The production of the macrophage cytokines, IL-1 and TNF-$\alpha$, by RAW 264.7 treated with WEP was examined from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml with dose dependent manner. Nitric oxide (NO) production was also increased when cells were exposed to combination of LPS and WEP from 2.5 $\mu\textrm{g}$/ml up to 25 $\mu\textrm{g}$/ml. At high dose of WEP (50 to 100 $\mu\textrm{g}$/ml) used to prescribe for anti-inflammatory and analgesic medicine showed inhibition of NO production in LPS-stimulated macrophage. Besides cytokine production, NO release, surface molecule expression and cell morphologic antigen expression were increased in response to the stimulation by WEP. These results suggested WEP may function through macrophage activation.

• Archives of Pharmacal Research
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• v.27 no.1
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• pp.94-98
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• 2004

Effects of dopaminergic drugs on the degranulation of mast cells (RBL-2H3 cells) and the nitric oxide production from macrophage cells (RAW 264.7) were studied. Among the dopaminergic agonists and antagonists tested, bromocriptine, 7-OH-DPAT, haloperidol, and clozapine showed potent inhibitions of mast cell degranualtion ($IC_{50} value, 5 \mu$ M). However, these dopaminergic agents did not affect the tyrosine phosphorylations of the signaling components of the high affinity IgE receptor ($Fc\varepsilonRI$), such as Syk, $PLC\gamma1$, and $PLC\gamma2$.; This suggested that these signaling components were not involved in the inhibition of the mast cell degranulation by these compounds. On the other hand, dopamine, bromocriptine, 7-OH-DAPT, and haloperidol markedly inhibited the nitric oxide production from RAW 264.7 cells ($IC_{50}$ values, 10-20$\mu$M). Bromocriptine, a dopamine agonist that is routinely used for the treatment of Parkinsons disease, inhibited the expression of the inducible nitric oxide synthase at an early stage of the LPS-induced protein expression in a dose-dependent manner. The results suggested that these dopaminergic agents, when used for the treatment of dopamine receptors-related diseases, such as Schizophrenia or Parkinsons disease, might have additional beneficial effects.

• Journal of Life Science
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• v.17 no.10
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• pp.1354-1360
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• 2007

Theaflavins (TF) and thearubigins (TR) are constituents of tea pigments which are polyphenols derived from Korean fermentation tea. After TF, TR and [(-) epigallocatechin-3-gallate](EGCG) have been applied to macrophage cell line (RAW264.7) nitric oxide (NO) synthesis and cytokines production were estimated. Cytokines production by enzyme linked immune-sorbent assay (ELISA) determined. NO production was increased by about 1.5-folds at the dose of $80\;{\mu}g/ml$ compared to control and lipopolysaccharide (LPS) stimulation when TF, TR and EGCG were applied to a RAW264.7 cell. Interleukin-6 (IL-6), Tumor necrosis factor ($TNF-{\alpha}$) and granulocyte-macrophage colony stimulating factor (GM-CSF) increased depended on concentrations of TF, TR and EGCG. The production of tumor necrosis $factor-{\alpha}$ increased highly in TR, TF and EGCG group with LPS. These results suggest that TF, TR and EGCG have immune-enhancement effect through the cytokine production. TF, TR and EGCG inhibited cancer cell viability, the anticancer effect of these polyphenols may explain the anti-tumor promotion action and antioxidant activity of these tea constituents.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.263-268
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• 2021

7-Hydroxy-4-methylcoumarin (7H-4MC) inhibits hyaluronan production in multiple cell lines and tissue types both in vitro and in vivo. It is a commercially available drug approved for human use, called hymecromone, in European and Asian countries to prevent biliary spasms. Nevertheless, as the pharmacological efficacy of 7H-4MC has not yet been reported in macrophages, this study investigated its anti-inflammatory effects and mechanism of action using lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. LPS-induced RAW 264.7 cells were treated with various concentrations of 7H-4MC (62.5, 125, 250, and 500 μM). The application of 7H-4MC significantly reduced nitric oxide and prostaglandin E₂ production without cytotoxic effects. Additionally, 7H-4MC strongly decreased the expression of inducible nitric oxide synthase and cyclooxygenase. Furthermore, 7H-4MC reduced the production of proinflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. Finally, 7H-4MC exerted its potent anti-inflammatory actions via the upregulation of IκB-α production, which led to the inhibition of nuclear factor-κB (NF-κB) activity. These results, obtained in macrophage cell lines, suggest that 7H-4MC prevents inflammatory diseases via the NF-κB signaling pathway and that its use could be beneficial for human health. Ultimately, this is the first report describing the anti-inflammatory activity of 7H-4MC in a macrophage cell line.

• Toxicological Research
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• v.27 no.3
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• pp.167-172
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• 2011

We demonstrate that glycoprotein isolated from Dioscorea batatas (GDB) activates macrophage function. Analysis of the infiltration of macrophages into peritoneal cavity showed GDB treatment significantly increased the recruitment of macrophages into the peritoneal cavity. In order to further confirm and investigate the mechanism of GDB on macrophage activation, we analyzed the effects of GDB on the cytokine expression including IL-$1{\beta}$, TNF-${\alpha}$, and IL-6 in mouse peritoneal macrophages. GDB increased the expression of IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. Cytokine induction by GDB was further confirmed by RT-PCR and ELISA in mouse macrophage cell line, RAW264.7 cells. Treatment of RAW264.7 cells with GDB produced strong induction of NF-${\kappa}B$ DNA binding and MAPK phosphorylation, markers for macrophage activation and important factors for cytokine gene expression. Collectively, this series of experiments indicates that GDB stimulates macrophage activation.

• 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.89-101
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• 2007

Objectives : This study was carried out to investigate the effects of Jeondo-San(JDS) on anti-Inflammation and anti-Propionibacterium acnes. Methods : The effects of JDS on anti-Inflammation and anti-Propionibacterium acnes 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 and the sterilizing power for Propionibacterium acnes. Results : 1. All concentrations of JDS has no cytotoxicity in Raw 264.7 cell. 2. All concentrations of JDS inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of JDS did not significantly inhibit the production of $TNF-{\alpha}$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of JDS inhibited the production of $PGE_2$ in the Raw 264.7 cell stimulated with LPS. 5. All concentrations of JDS did not inhibit the expression of COX-2 but concentrations of 50\;{\mu}g/ml$, 100\;{\mu}g/ml$ JDS inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. Concentrations of 50\;{\mu}g/ml$, 100\;{\mu}g/ml$ JDS has the effect of blocking $NF-{\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell. 7. All concentrations of IDS did not have the inhibitory effect of Propionibactrium acnes. Conclusions : The present date suggest that JDS has a effect on the stage of inflammation of acne.

• Biomolecules & Therapeutics
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• v.23 no.5
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• pp.407-413
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• 2015

Paraquat dichloride (N,N-dimethyl-4-4'-bipiridinium, PQ) is an extremely toxic chemical that is widely used in herbicides. PQ generates reactive oxygen species (ROS) and causes multiple organ failure. In particular, PQ has been reported to be an immunotoxic agrochemical compound. PQ was shown to decrease the number of macrophages in rats and suppress monocyte phagocytic activity in mice. However, the effect of PQ on macrophage cell viability remains unclear. In this study, we evaluated the cytotoxic effect of PQ on the mouse macrophage cell line, RAW264.7 and its possible mechanism of action. RAW264.7 cells were treated with PQ (0, 75, and $150{\mu}M$), and cellular apoptosis, mitochondrial membrane potential (MMP), and intracellular ROS levels were determined. Morphological changes to the cell nucleus and cellular apoptosis were also evaluated by DAPI and Annexin V staining, respectively. In this study, PQ induced apoptotic cell death by dose-dependently decreasing MMP. Additionally, PQ increased the cleaved form of caspase-3, an apoptotic marker. In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway. Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.