• The Korean Journal of Physiology and Pharmacology
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• v.1 no.2
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• pp.209-219
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• 1997

Previous studies have demonstrated that oxidative stress involving generation of reactive oxygen species (ROS) is responsible for the cytotoxic action of $TNF{\alpha}$. Protective effect of small heat shock proteins (small HSP) against diverse oxidative stress conditions has been suggeted. Although overexpression of small hsp was shown to provide an enhanced survival of $TNF{\alpha}$-sensitive cells when challenged with $TNF{\alpha}$, neither the nature of $TNF{\alpha}$-induced cytotoxicity nor the protective mechanism of small HSP has not been completely understood. In this study, we have attempted to determine whether $TNF{\alpha}$ induces oxidative DNA damage in $TNF{\alpha}$-sensitive L929 cells. We chose to measure the level of 8-hydroxy-2'-deoxyguanosine (8 ohdG), which has been increasingly recognized as one of the most sensitive markers of oxidative DNA damage. Our results clearly demonstrated that the level of 8 ohdG increased in L929 cells in a $TNF{\alpha}$ dose-dependent manner. Subsequently, we asked whether small HSP has a protective effect on $TNF{\alpha}$-induced oxidative DNA damage. To accomplish this goal, we have stably transfected L929 cells with mouse small hsp cDNA (hsp25) since these cells are devoid of endogenous small hsps. We found that $TNF{\alpha}$-induced 8 ohdG was decreased in cells overexpressing exogenous small hsp. We also found that the cell killing activity of $TNF{\alpha}$ was decreased in these cells as measured by clonogenic survival. Taken together, results from the current study show that cytotoxic mechanism of $TNF{\alpha}$ involves oxidative damage of DNA and that overexpression of the small hsp reduces this oxidative damage. We suggest that the reduction of oxidative DNA damage is one of the most important protective mechanisms of small HSP against $TNF{\alpha}$.

• Korean Journal of Medicinal Crop Science
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• v.15 no.1
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• pp.30-37
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• 2007

It is well-known that Korean mistletoe (Viscum album) extract has an immune activity and anticancer effect. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to examine whether this extract might activate human peripheral monocyte to produce tumor necrosis $factor-\alpha$ $(TNF-\alpha)$. To examine the effect of M11C on the production of $TNF-\alpha$ from monocyte, the monocyte were stimulated by the M11C, and then collected the supernatant (M11C stimulated monocyte-conditioned media; MCM). MCM was treated to the $TNF-\alpha$ sensitive L929 cells, and then L929 cytotoxicity was measured by means of MTT. MCM had cytotoxic effect on L929. And the cytotoxic effect of MCM on L929 was almost abolished by $anti-TNF-\alpha$ antibody. These data indicated that MCM contained $TNF-\alpha$, suggesting the $TNF-\alpha$ generation from M11C-stimulated monocyte. This suggestion was confirmed from the data that $TNF-\alpha$ was highly detected in MCM by immunoblotting technique. M11C effect on $TNF-\alpha$ production from monocyte was in the dose and stimulating time dependent manners. Also the effect of M11C on the expression of $TNF-\alpha$ mRNA from monocyte was shown in the dose and stimulating time dependent manners. As a result, Korean mistletoe extract, M11C, could be used for an immunostimulator.

• 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.

• The Journal of Internal Korean Medicine
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• v.25 no.1
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• pp.28-45
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• 2004

Objectives : The main purpose of this study is to evaluate the effect of Injinchunggan-tang on $TNF-{\alpha}$ signal transmission system. Materials and Methods : We analyzed the following with quantitative RT-PCR method; the effect of Injinchunggan-tang on secretion of $TNF-\alpha$ mRNA/protein and stability, the effect on gene revelation that consists of signal transmission system (TRAIL, NIK, A20, TRADD, RAIDD, RIP TNFR-I, TNFR-II, TRAF1, TRAF2, FADD), the one on activation of p38, Erk1/2 MAPK and the rate of nuclear $NF-{\kappa}B/cytosolic\;NF-{\kappa}B$ in HepG2 cell. We also analyzed the inhibitory effect of Injinchunggan-tang on the apoptosis of HepG2 cell that $TNF-{\alpha}$ induces and the $NF-{\kappa}B$ restraint effected by transfection of $I{\kappa}B{\Delta}N$ through tryphan blue exclusion assay. Results : Injinchunggan-tang prohibits revelation of $TNF-{\alpha}$ mRNA in HepG2 cell and the creation of protein. However, it has no effect on the stability of $TNF-{\alpha}$ mRNA. While it did not have any effect on the generation of TRAIL, NIK, A20, TRADD, RAIDD and RIP genes, Injinchunggan-tang reduces the revelation of TNFR-I, TNFR-II, TRAF1, TRAF2 and FADD genes. It has been confirmed that Injinchunggan-tang restraints the revelation of $TNF-{\alpha}$ mRNA that is promoted by ethanol, acetaldehyde, lipopolysaccharide, in proportion to the treatment density and time. It activated $NF-{\kappa}B$ of HepG2 cell and promoted activation of $NF-{\kappa}B$ that is occurred by $TNF-{\alpha}$. It has been observed that the restraint effect against the $TNF-{\alpha}$ inducing apoptosis is lost when it is intercepted the function of $NF-{\kappa}B$ in HepG2 cell. Conclusion: It has been confirmed that Injinchunggan-tang has restraining effect against the revelation of $TNF-{\alpha}$ and mRNA that is constituent element of TNF-a signal transmission system. It also has been revealed that it restraints the activation of p38, Erk1/2 by $TNF-{\alpha}$. Through this prohibiting effect, it is inferred that it restraints signal transmission among various cells that are related to inflammation reaction. Meanwhile, Injinchunggan-tang protects liver cell from apoptosis that is caused by $TNF-{\alpha}$, by maintaining the activating function for $NF-{\kappa}B$.

• The Journal of Internal Korean Medicine
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• v.20 no.1
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• pp.143-152
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• 1999

Dansam, the root of Salvia miltiorrhiza Bge, (Labiatae), has a bitter taste and a slightly 'cold' property, and is nontoxic. In the present study, effect of Dansam on nitric oxide (NO) generation from peritoneal macrophags was examined. Dansam had no effect on NO generation by itself, whereas recombinant interferon-${\gamma}\;(rIFN-{\gamma})$ alone had modest activity. When Dansam was used in combination with $rIFN-{\gamma}$, there was a marked cooperative induction of NO generation in a dose-dependent manner, The optimal effect of Dansam on NO generation was shown at 6 hr after treatment with $rIFN-{\gamma}$. Furthermore, the effect of Dansam was mainly dependent on Dansam-induced tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ secretion. These results suggest that Dansam induces NO generation from macrophages by the result of Dansam-induced $TNF-{\alpha}$ secretion.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.103-112
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• 2008

Objectives : This research aimed to study the cytotoxicity and immuno modulating activity of fermented Artemisia argyi Lev. et Vant.(Compositae). Methods : Effect of fermented Artemisiae Argyi Folium extracts, which were fermented by Sacchromyces cerevisiae STV89(AFS), on cell viability, generation of ROS within cells, generation of NO and the level of cytokines($TNF-{\alpha}$ and IL-6) was measured using mouse macrophage RAW 264.7 cell. Results : 1. Result of MTT assay conducted to verify the cytotoxicity of fermented Artemisiae argyi folium extract illustrated that, when fermented Artemisiae argyi folium extract was processed for each concentration, there was no excessive induction of cytoxicity in the RAW 264.7 cell. 2. Fermented Artemisiae Argyi Folium extract increased the generation of H2O2 within RAW 264.7 cell as well as significantly increased inhibition of generation of H2O2 in macrophage induced by LPS. 3. Fermented Artemisiae Argyi Folium extract inhibited generation of NO in RAW 264.7 cell, and significantly inhibited increase in generation of NO of macrophage induced by LPS. 4. Fermented Artemisiae Argyi Folium extract, AFS has significantly reduced the increase in the generation of $TNF-{\alpha}$ above 10 ${\mu}g/mL$. 5. Fermented Artemisiae Argyi Folium extract, AFS has significantly reduced the increase in generation of IL-6 above 50 ${\mu}g/mL$. Conclusions : AFS fermented extract produced from Artemisiae Argyi Flium, have increased generation of ROS and reduced generation of NO in RAW 264.7 cell without excessively inducing cytotoxicity of RAW 264.7 cell. In addition, they displayed significant immuno modulating activities including inhibition of generation of $TNF-{\alpha}$ and IL-6 in macrophage, induced by LPS.

• BMB Reports
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• v.44 no.7
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• pp.462-467
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• 2011

Up-regulation of selected matrix metalloproteinases (MMPs) such as MMP-9 contributes to inflammatory processes during the development of various skin diseases, such as atopic dermatitis. In this study, we examined the effect of a cell-permeable superoxide dismutase (Tat-SOD) on TNF-${\alpha}$-induced MMP-9 expression in human keratinocyte cells (HaCaT). When Tat-SOD was added to the culture medium of HaCaT cells, it rapidly entered the cells in dose- and time-dependent manners. Tat-SOD decreased TNF-${\alpha}$-induced reactive oxygen species (ROS) generation. Tat-SOD also inhibited TNF-${\alpha}$-induced NF-${\kappa}B$ DNA binding activity. Treatment of HaCaT cells with Tat-SOD significantly inhibited TNF-${\alpha}$-induced mRNA and protein expression of MMP-9, as measured by RT-PCR and Western blot analysis. In addition, Tat-SOD suppressed TNF-${\alpha}$-induced gelatinolytic activity of MMP-9. Taken together, our results indicate that Tat-SOD can suppress TNF-${\alpha}$-induced MMP-9 expression via ROS-NF-${\kappa}B$-dependent mechanisms in keratinocytes, and therefore can be used as an immunomodulatory agent against inflammatory skin diseases related to oxidative stress.

• The Korea Journal of Herbology
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• v.21 no.1
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• pp.89-100
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• 2006

Objectives : The effects of methanol extracts from the cortex of Quercus dentata Thunb. and Quercus acutissima Carruth, on the activation of macrophage were examined. Methods : Methanol extracts of Quercus dentata (QD) and Quercus acutissima (QA) were applied to cell line RAW264,7 (macrophage), and their effects were examined. Results : 1. Extracts from QD and QA had no specific influence on the cell growth. 2. Extracts from QD and QA did not activate macrophage independently, but the addition of $IFN-{\gamma}$ facilitated the generation of macrophage's nitric oxide(NO). 3. QD and QA extracts increased the manifestation of iNOS gene, when macrophage was activated by $IFN-{\gamma}$. 4. QD and QA extracts increased the manifestation of $TNF-{\alpha}$ in macrophage, which took 2 hours. 5. QD and QA extracts increased the generation of $PGE_2$ in macrophage. Conclusion : QD and QA activate macrophage in the presence of $IFN-{\gamma}$. After activation is primarily facilated by $IFN-{\gamma}$, it works on macrophage secondarily for the manifestation of iNOS gene and for the generation of $TNF-{\alpha}$.

• BMB Reports
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• v.42 no.5
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• pp.265-270
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• 2009

Due to their multiple biological activities, flavonoids have gained attention as potentially useful therapeutics for a variety of diseases including cancer, cardiovascular diseases, and autoimmune diseases. In this study, we demonstrated that several flavonoids, including kaempferol, quercetin, fisetin, and chrysin block TNF-$\alpha$ induced IL-8 promoter activation and gene expression in HEK 293 cells. In addition, phosphorylation and degradation of $I{\kappa}B{\alpha}$ and translocation of NF-${\kappa}B$ p65 were inhibited by these flavonoids in TNF-$\alpha$-stimulated HEK 293 cells. Furthermore, generation of reactive oxygen species (ROS) in response to TNF-$\alpha$ was reduced by the flavonoids. Moreover, although pretreatment with fisetin, quercetin, or chrysin decreased cell viability, kaempferol did not. Taken together, these findings suggest that kaempferol would be useful for the treatment of TNF-$\alpha$-induced inflammatory diseases.

• Korean Journal of Pharmacognosy
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• v.34 no.3 s.134
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• pp.223-227
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• 2003

In the present study, anti-inflammatory activity of amygdalin isolated from persicae Semen have been evaluated on lipopolysaccharide (LPS)-induced release of nitric oxide (NO), prostaglandin $E_2\;(PGE_2)$ and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) by the macrophage RAW 264.7 cells. Amygdalin significantly inhibited generation of NO and $TNF-{\alpha}$ on LPS-stimulated RAW264.7 cells in a concentration-dependent manner. Consistent with these observations, the expression of inducible NO synthase (iNOS) enzyme was also inhibited by amygdalin in a concentration-dependent manner. However, amygdalin did not show any influence on the synthesis of $PGE_2$ and the expression of COX-2. Thus, this study suggests that amygdalin-mediated inhibition of iNOS expression, and $TNF-{\alpha}$ release may be one of the mechanisms responsible for the anti-inflammatory effects of Persicae Semen.