• Journal of Nutrition and Health
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• v.49 no.5
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• pp.288-294
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• 2016

Purpose: The aim of this work was to investigate the beneficial effects of rhamnazin against inflammation, reactive oxygen species (ROS)/reactive nitrogen species (RNS), and anti-oxidative activity in murine macrophage RAW264.7 cells. Methods: To examine the beneficial properties of rhamnazin on inflammation, ROS/ RNS, and anti-oxidative activity in the murine macrophage RAW264.7 cell model, several key markers, including COX and 5-LO activities, $NO^{\cdot}$, $ONOO^-$, total reactive species formation, lipid peroxidation, $^{\cdot}O_2$ levels, and catalase activity were estimated. Results: Results show that rhamnazin was protective against LPS-induced cytotoxicity in macrophage cells. The underlying action of rhamnazin might be through modulation of ROS/RNS and anti-oxidative activity through regulation of total reactive species production, lipid peroxidation, catalase activity, and $^{\cdot}O_2$, $NO^{\cdot}$, and $ONOO^{\cdot}$ levels. In addition, rhamnazin down-regulated the activities of pro-inflammatory COX and 5-LO. Conclusion: The plausible action by which rhamnazin renders its protective effects in macrophage cells is likely due to its capability to regulate LPS-induced inflammation, ROS/ RNS, and anti-oxidative activity.

• Proceedings of the Korean Vacuum Society Conference
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• 2015.08a
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• pp.159-159
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• 2015

A new approach for antimicrobial is based on the overproduction of reactive nitrogen species (RNS), especially; nitric oxide (NO) and peroxinitrite (ONOO-) are important factors to deactivate the bacteria. Recently, non-thermal atmospheric pressure plasma jet (APPJ) has been frequently used in the field of microbial sterilization through the generation of different kinds of RNS/ROS species. However, in previous study we showed APPJ has combine effects ROS/RNS on bacterial sterilization. It is not still clear whether this bacterial killing effect has been done through ROS or RNS. We need to further investigate separate effect of ROS and RNS on bacterial sterilization. Hence, in this work, we have enhanced NO production, especially; by applying a 1% of HNO3 vapour to the N2 based APPJ. In comparison with nitrogen plasma with inclusion of water vapour plasma, it has been shown that nitrogen plasma with inclusion of 1% of HNO3 vapour has higher efficiency in killing the E. coli through the high production of NO. We also investigate the enhancement of NO species both in atmosphere by emission spectrum and inside the solution by ultraviolet absorption spectroscopy. Moreover, qPCR analysis of oxidative stress mRNA shows higher gene expression. It is noted that 1% of HNO3 vapour plasma generates high amount of NO for killing bacteria.

• International Journal of Oral Biology
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• v.41 no.3
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• pp.141-147
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• 2016

Reactive oxygen species (ROS) and nitrogen species (RNS) are both important signaling molecules involved in pain transmission in the dorsal horn of the spinal cord. Xanthine oxidase (XO) is a well-known enzyme for the generation of superoxide anions ($O_2^{\bullet-}$), while S-nitroso-N-acetyl-DL-penicillamine (SNAP) is a representative nitric oxide (NO) donor. In this study, we used patch clamp recording in spinal slices of rats to investigate the effects of $O_2^{\bullet-}$ and NO on the excitability of substantia gelatinosa (SG) neurons. We also used confocal scanning laser microscopy to measure XO- and SNAP-induced ROS and RNS production in live slices. We observed that the ROS level increased during the perfusion of xanthine and xanthine oxidase (X/XO) compound and SNAP after the loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), which is an indicator of intracellular ROS and RNS. Application of ROS donors such as X/XO, ${\beta}-nicotinamide$ adenine dinucleotide phosphate (NADPH), and 3-morpholinosydnomimine (SIN-1) induced a membrane depolarization and inward currents. SNAP, an RNS donor, also induced membrane depolarization and inward currents. X/XO-induced inward currents were significantly decreased by pretreatment with phenyl N-tert-butylnitrone (PBN; nonspecific ROS and RNS scavenger) and manganese(III) tetrakis(4-benzoic acid) porphyrin (MnTBAP; superoxide dismutase mimetics). Nitro-L-arginine methyl ester (NAME; NO scavenger) also slightly decreased X/XO-induced inward currents, suggesting that X/XO-induced responses can be involved in the generation of peroxynitrite ($ONOO^-$). Our data suggest that elevated ROS, especially $O_2^{\bullet-}$, NO and $ONOO^-$, in the spinal cord can increase the excitability of the SG neurons related to pain transmission.

• Proceedings of the Korean Vacuum Society Conference
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• 2016.02a
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• pp.230.2-230.2
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• 2016

A new approach for antimicrobial is based on the overproduction of reactive nitrogen species (RNS), especially; nitric oxide (NO) and peroxinitrite ($ONOO^-$-) are important factors to deactivate the bacteria. Recently, non-thermal atmospheric pressure plasma jet (APPJ) has been frequently used in the field of microbial sterilization through the generation of different kinds of RNS/ROS species. However, in previous study we showed APPJ has combine effects ROS/RNS on bacterial sterilization. It is not still clear whether this bacterial killing effect has been done through ROS or RNS. We need to further investigate separate effect of ROS and RNS on bacterial sterilization. Hence, in this work, we have enhanced NO production, especially; by applying a 1% of HNO3 vapour to the N2 based APPJ. In comparison with nitrogen plasma with inclusion of water vapour plasma, it has been shown that nitrogen plasma with inclusion of 1% of HNO3 vapour has higher efficiency in killing the E. coli and different type of cancer cell through the high production of NO. We also investigate the enhancement of NO species both in atmosphere by emission spectrum and inside the solution by ultraviolet absorption spectroscopy. Moreover, qPCR analysis of oxidative stress mRNA shows higher gene expression. It is noted that 1% of HNO3 vapour plasma generates high amount of NO for killing bacteria and cancer cell killing.

• Journal of Korean Ophthalmic Optics Society
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• v.13 no.3
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• pp.103-109
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• 2008

Purpose: This review illustrates an importance of oxidative stress caused by reactive oxygen species (ROS) and reactive nitrogen species (RNS) generation in association with eye disease, especially of cataract, and discusses an important role of lipid peroxide as a mediator of oxidative stress-related ocular dysfunction. Methods: Oxidative stress, resulted from the cellular production of ROS and RNS, is known to cause various forms of cellular damages such as protein oxidation, DNA breaks, apoptosis, and lipid peroxidation. These damages can be developed to human diseases. Accumulating evidence strongly suggests that continuous or constant exposure of eye tissues to oxidative stress is a main cause of cataractogenesis. Therefore, we investigated the action of oxidative stress in ocular dysfunction. Results: The ocular lens is continuously attacked by ROS inevitable generated from the process of cellular metabolism and the chronic exposure to ultraviolet. Excessive generation of ROS, resulting in degradation, oxidation, crosslinking and aggregation of lens proteins, is regarded as an important factor in development of cataract. Conclusions: These oxidative stress and oxidant/antioxidant imbalance produces the excess ROS which can lead to eye dysfunction. Even though known results, it should be noted that there is limited information on the molecular mechanism which can be better defined with the interrelation of oxidative stress and optic abnormalities.

• Herbal Formula Science
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• v.22 no.1
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• pp.79-92
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• 2014

Objectives : The aim of this study was identification of the anti-oxidative and anti-inflammatory effects of Talmyung-san (TMS) in mouse macrophage, RAW264.7 cells. Methods : To identify the anti-oxidative effect of TMS, scavenging activities of DPPH radical, nitric oxide and peroxynitrite were measured in vitro. In RAW264.7 cells, DCFH-DA assay was conducted to examine the inhibitory effect of TMS on ROS production in response to lipopolysaccharide. And the productions of nitric oxide (NO), $PGE_2$ and pro-inflammatory cytokines were measured. The levels of COX-2, iNOS, nuclear NF-${\kappa}B$ p65 expression and phosphorylation of $I{\kappa}B-{\alpha}$ in cytosol were detected by western blotting analyses. Results : TMS couldn't scavenged DPPH radical, but nitric oxide and peroxynitrite were decreased. TMS decreased intracellular ROS, NO, and IL-$1{\beta}$ production effectively. However, TMS inhibited $PGE_2$ levels only in high concentration ($300{\mu}g/m{\ell}$) and TMS failed to suppress the production of IL-6 and TNF-${\alpha}$. Results from immunoblot analyses revealed that TMS decreased activation of COX-2, iNOS, phosphorylation of $I{\kappa}B-{\alpha}$ and nuclear translocation of p65. Conclusions : TMS has anti-RNS and anti-inflammatory effects via NF-${\kappa}B$ pathway and more intensive studies will be required to evaluate therapeutic potential of TMS.

• Tuberculosis and Respiratory Diseases
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• v.52 no.6
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• pp.616-626
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• 2002

Background : The present study was performed to further improve our understanding of molecular mechanisms involved in the activation of NFkB, a major transcriptional factor involved in the inflammatory response in the lung, by particulate matter in lung epithelial cells with an aerodynamic diameter of less than $2.5{\mu}m$(PM2.5). Materials and Methods : Immediate production of reactive oxygen species (ROS) and nitrogen species (RNS), with the PM2.5 induced expression of inducible nitric oxide synthase (iNOS), $I{\kappa}B$ degradation and $NF{\kappa}B$-dependent transcriptional activity, in 549 cells, were monitored. Addition, we also examined the effect of the iNOS inhibitor, L-N6-(1-iminoethyl) lysine hydrochloride (L-NIL), on the PM2.5-induced $NF{\kappa}B$ activation in A549 cells. Results : The rapid degradation of $I{\kappa}B$ and the increase of transcriptional activity of the $NF{\kappa}B$-dependent promotor were observed in A549 cells exposed to PM2.5. The immediate production of ROS in response to PM2.5 in A549 cells was not clearly detected, although immediate responses were observed in RAW264.7 cells. A 549 cells, cultured in the presence of PM2.5, produced an increase in NO, which was noticeably significant after 15 min of exposure with the expression of iNOS mRNA. The addition of L-NIL, an iNOS inhibitor, significantly inhibited the PM2.5-induced $I{\kappa}B$ degradation and the increase of the $NF{\kappa}B$-dependent transcriptional activity. Conclusion : These results suggest that PM2.5 stimulates the immediate production of RNS, leading to the activation of $NF{\kappa}B$ in the pulmonary epithelium.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1320-1331
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• 2009

Oxidative stress and inflammation are important events in the development of chronic inflammatory diseases including arthritis, atherosclerosis, diabetes, hypertension. Cynomorium songaricum (CS) has been used as a traditional Korean herbal medicine, and it is currently used in traditional clinics to treat frequent urination, spermatorrhea, weakness of the sinews and constipation in the folk medicine. The aim of this study was to determine whether fractionated extracts of CS inhibit free radical generation such as DPPH radical, superoxide radical, nitric oxide and peroxynitrite, production of nitrite an index of NO, $PGE_2$, iNOS, COX-2 and pro-inflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Our results indicated that the most superior extract which scavenged DPPH radical, reactive oxygen species (ROS) and RNS was CS ethyl acetate extract (CSEA). Moreover, CSEA significantly inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-$\alpha$, IL-$1{\beta}$ and IL-6 formation in macrophages. Furthermore, CSEA treatment also blocked LPS-induced intracellular ROS production and the activation of NF-${\kappa}B$. These findings indicate that CSEA inhibits the production of pro-inflammatory mediators and cytokines via the suppression of ROS production and NF-${\kappa}B$ activation. Take together, these results indicate that CSEA has the potential for use as an natural anti-oxidant and an agent of anti-chronic inflammatory diseases.

• Preventive Nutrition and Food Science
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• v.17 no.2
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• pp.122-128
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• 2012

Gelidium (G.) elegans is a red alga inhabiting intertidal areas of North East Asia. We examined anti-oxidative and anti-inflammatory effects of G. elegans, depending on drying and extraction conditions, by determining reactive oxygen species (ROS) and nitric oxide (NO) in 3T3-L1 and RAW 264.7 cells. Extraction yields of samples using hot air drying (HD) and far-infrared ray drying (FID) were significantly higher than those using natural air drying (ND). The 70% ethanol extracts showed the highest total phenol and flavonoid contents compared to other extracts (0, 30, and 50% ethanol) under tested drying conditions. The scavenging activity on 2,2-diphenyl- 1-picrylhydrazyl (DPPH) and nitrite correlated with total phenol or flavonoid content in the extracts. The greatest DPPH scavenging effect was observed in 70% ethanol extract from FID and HD conditions. The production of ROS and NO in 3T3-L1 and macrophage cells greatly decreased with the 70% ethanol extraction derived from FID. This study suggests that 70% ethanol extraction of G. elegans dried by FID is the most optimal condition to obtain efficiently antioxidant compounds of G. elegans.

• Journal of Life Science
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• v.19 no.5
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• pp.598-606
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• 2009

Oxidative stress by free radicals is a major cause of neuronal cell death. Excitotoxicity in hypoxia/ischemia causes an increase in reactive oxygen species (ROS) and a loss of mitochondrial membrane potential (MMP), resulting in dysfunction of the mitochondria and cell death. Pinelliae Rhizoma (PR) is a traditional medicine for incipient stroke. We investigated the effects of PR Water-Extract on the modulation of ROS and MMP in a hypoxic model using cultured rat cortical cells. PR Water-Extract was added to the culture medium at various concentrations (0.25${\sim}$5, 5.0 ${\mu}g/ml$) on day in vitro 12(DIV12), given a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 hr), and cell viability was assessed on DIV15 by Lactate Dehydrogenase Assay (LDH assays). PR Water-Extract showed a statistically significant effect on neuroprotection (10${\sim}$15% increase in viability; p<0.01) at 1.0 and 2.5 ${\mu}g/ml$ in normoxia and hypoxia. Measurement of ROS production by $H_2DCF-DA$ stainings showed that PR Water-Extract efficiently reduced the number and intensity of ROS-producing neurons, especially at 1 hr post shock and DIV15. When MMP was measured by JC-1 stainings, PR Water-Extract efficiently maintained high-energy charged mitochondria. These results indicate that PR Water-Extract protects neurons in hypoxia by preventing ROS production and preserving the cellular energy level.