• The Journal of Korean Medicine
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• v.27 no.2 s.66
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• pp.171-181
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• 2006

Objectives : Peroxynitrite($ONOO^-$), superoxide anion(${\cdot}{O_2}^-$) and nitric oxide (NO) is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate scavenging activities for $ONOO^-$ and its precursors, NO and ${\cdot}{O_2}^-$ of Vespae Nidus. Methods : Dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used to investigate scavenging activities of $ONOO^-,\;NO,\;{\cdot}{O_2}^-$. Six-months-old ICR mice were used. After mice were injected with lipopolysaccharides(LPS), kidney organization was evaluated. Three comparison groups of ICR mice were used : a normal group, an experimental group that was fed Vespae Nidus extract and then injected with LPS, and a control group that was injected with LPS. Scavenging activities of $ONOO^-,\;NO,\;{\cdot}{O_2}^-$ in these groups were measured in the same way. Results : Vespae Nidus markedly scavenged authentic $ONOO^-,\;{\cdot}{O_2}^-$ and NO. It also inhibited $ONOO^-$ induced by ${\cdot}{O_2}^-$ and NO which are derived trom SIN-1. Furthermore, it inhibited $ONOO^-,\;{\cdot}{O_2}^-$, and NO generation by Vespae Nidus in LPS-treated ICR mouse kidney postmitochondria. Conclusions : These results suggest that Vespae Nidus might be developed as an effective $ONOO^-,\;{\cdot}{O_2}^-$, and NO scavenger for the prevention of the aging process and age-related diseases.

• Journal of the Korean Chemical Society
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• v.34 no.2
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• pp.189-196
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• 1990

8-Acyl-2-hydroxyphenazine-5,10-dioxides and 8-acyl-2-aminophenazine-5,10-dioxides were synthesized by the reaction of hydroquinone and 4-aminophenol with 6-acylbenzofuroxans which had been obtained from acetanilide and n-acyl chlorides bearing butanoyl, hexanoyl and octanoyl groups. The antimicrobial activities of these phenazine dioxide derivatives were investigated in terms of minimum inhibitory concentration by the common twofold dilution technique. It was observed that the antimicrobial activity of the phenazine dioxide derivatives bearing octanoyl group was stronger than that of those bearing butanoyl and hexanoyl groups in gram positive microorgamisms, but it was observed that the antimicrobial activity and the number of the carbon atom of acyl groups did not have any relation in gram negative microorganisms. When the activity of xanthine oxidase which is the key enzyme in the generation of superoxide anion radical ($O_2^-$), was measured in the presence of phenazine dioxide derivatives, the inhibitory action of the enzyme activity of 8-acyl-2-hydroxyphenazine-5,10-dioxides was increased in accordance with the number of the carbon atom of acyl groups.

• The Journal of Internal Korean Medicine
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• v.24 no.1
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• pp.75-83
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• 2003

Objectives : Peroxynitrite($ONOO^-$), formed from the reaction of $O2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been involved in the aging process and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_{2^-}$ and its scavenging mechanism of Zingiberis Rhizoma (ZR). Methods : To investigate scavenging activities of $ONOO^-,\;NO,\;O_{2^-}$ and its scavenging mechanism, we used fluorescent probes like DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on ZR was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. The scavenging efficacy was expressed as IC50, showing the concentration of each sample that is required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of ZR on $ONOO^-$-induced nitration of bovine serum albumin was investigated through immuno-assay with a monoclonal anti-nitryrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results : ZR markedly scavenged authentic $ONOO^-,\;O_{2^-}$ and NO. It also inhibited $ONOO^-$ induced by $O_{2^-}$ and NO which are derived from SIN-1. The data demonstrated that ZR led to decreased $ONOO^-$ mediated nitration of tyrosine through electron donation. It also inhibited the nitration of bovine serum albumin induced by $ONOO^-$ in a dose-dependent manner. Furtheremore, it blocked LPS-induced ROS and RNS generation. Conclusions : These results suggest that ZR can be developed as an effective $ONOO^-$ scavenger for the prevention of aging process and age-related diseases.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.138-147
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• 2021

The overproduction of reactive nitrogen species (RNS) and reactive oxygen species (ROS) causes oxidative damage to neuronal cells, leading to the progression of neurodegenerative diseases. In this study, we determined the nitric oxide radical (NO), hydroxyl radical (·OH), and superoxide anion radical (O₂^-) scavenging activities of apigenin. Our results showed that apigenin exhibited remarkable, concentration-dependent ·OH, O₂^-, and NO radical scavenging activities. Particularly, apigenin indicated the strongest ·OH radical scavenging activity with 93.38% in the concentration of 100 µM. Furthermore, we also investigated the protective effects of apigenin against hydrogen peroxide (H₂O₂)-induced oxidative stress in SH-SY5Y cells. The H₂O₂ treatment resulted in a significant decrease in cell viability, as well as an increase in lactate dehydrogenase (LDH) release and ROS production compared with the H₂O₂-nontreated SH-SY5Y cells. However, the cell viability significantly increased in the apigenin-treated group, as well as inhibited ROS generation and LDH release compared with the H₂O₂-induced control group. To elucidate the protective mechanisms of apigenin against oxidative stress in SH-SY5Y, we analyzed the apoptosis-related protein expression. The apigenin treatment resulted in the downregulated expression of apoptosis-related protein markers, such as cytochrome C, cleaved caspase-3, poly (ADP)-ribose polymerase (PARP), and B-cell lymphoma 2-associated X (Bax), as well as the upregulated expression of anti-apoptosis markers such as B-cell lymphoma 2 (Bcl-2). In this study, we report that apigenin exhibits a neuroprotective effect against oxidative stress in SH-SY5Y cells. These results suggest that apigenin may be considered as a potential agent for neurodegenerative disease prevention.

• Toxicological Research
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• v.33 no.1
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• pp.43-48
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• 2017

With ultraviolet and visible light exposure, some pharmaceutical substances applied systemically or topically may cause phototoxic skin irritation. The major factor in phototoxicity is the generation of reactive oxygen species (ROS) such as singlet oxygen and superoxide anion that cause oxidative damage to DNA, lipids and proteins. Thus, measuring the generation of ROS can predict the phototoxic potential of a given substance indirectly. For this reason, a standard ROS assay (ROS assay) was developed and validated and provides an alternative method for phototoxicity evaluation. However, negative substances are over-predicted by the assay. Except for ultraviolet A (UVA), other UV ranges are not a major factor in causing phototoxicity and may lead to incorrect labeling of some non-phototoxic substances as being phototoxic in the ROS assay when using a solar simulator. A UVA stimulator is also widely used to evaluate phototoxicity in various test substances. Consequently, we identified the applicability of a UVA simulator to the ROS assay for photoreactivity. In this study, we tested 60 pharmaceutical substances including 50 phototoxins and 10 non-phototoxins to predict their phototoxic potential via the ROS assay with a UVA simulator. Following the ROS protocol, all test substances were dissolved in dimethyl sulfoxide or sodium phosphate buffer. The final concentration of the test solutions in the reaction mixture was 20 to $200{\mu}M$. The exposure was with $2.0{\sim}2.2mW/cm^2$ irradiance and optimization for a relevant dose of UVA was performed. The generation of ROS was compared before and after UVA exposure and was measured by a microplate spectrophotometer. Sensitivity and specificity values were 85.7% and 100.0% respectively, and the accuracy was 88.1%. From this analysis, the ROS assay with a UVA simulator is suitable for testing the photoreactivity and estimating the phototoxic potential of various test pharmaceutical substances.

• The Journal of Korean Medicine
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• v.30 no.1
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• pp.51-63
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• 2009

Objectives: Peroxynitrite ($ONOO^-$), superoxide anion radical (${\cdot}{O_2}^-$ and nitric oxide (NO) are cytotoxic because they can oxidize several cellular components such as proteins, lipids and DNA. They have been implicated in the aging processes, and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer, diabetes, obesity and atherosclerosis. The aim of this study was to investigate the $ONOO^-$, NO, ${\cdot}{O_2}^-$ scavenging and NF-${\kappa}B$ related anti-inflammatory activities of Sotosaja-hwan in ob/ob mice. Methods: Mice were grouped and treated for 5 weeks as follows. Both the normal lean (C57/BL6J black mice) and control obese (ob/ob mice) groups have received standard chow. The experimental groups were fed with a diet of chow supplemented with 30 and 90 mg Sotosaja-hwan per 1 kg of body weight for 14 days. For this study, the fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Western blotting was performed using anti-phospho-$I{\kappa}B$-${\alpha}$, anti-IKK-${\alpha}$, anti-NF-${\kappa}B$ (p50, p65), anti-COX-2, anti-iNOS, anti-YCAM-1 and anti-MMP-9 antibodies, respectively. Results: Sotosaja-hwan inhibited the generation of $ONOO^-$, NO and ${\cdot}{O_2}^-$ in the lipopolysaccharide (LPS)-treated mouse kidney postmitochondrial fraction in vitro. The generation of $ONOO^-$, NO, ${\cdot}{O_2}^-$ and PGE2 were inhibited in the Sotosaja-hwan-administered ob/ob mice groups. The GSH/GSSG ratio was decreased in the ob/ob mice, whereas the ratio was improved in the Sotosaja-hwan-administered groups. Sotosaja-hwan inhibited the protein expression levels of phospho-$I{\kappa}B$-${\alpha}$, IKK-${\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, YCAM-1 and MMP-9 genes. Conclusions: These results suggest that Sotosaja-hwan is an effective $ONOO^-$, ${\cdot}{O_2}^-$ and NO scavenger and has NF-kB related anti-inflammatory activity in ob/ob mice. Therefore, Sotosaja-hwan might be a potential therapeutic drug against the inflammation process and inflammation-related diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.30 no.4
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• pp.242-249
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• 2016

This study was to investigate the anti-inflammatory effects of Prunellae Herba(PH). The generation of superoxide anion radical (․O₂-), nitric oxide (NO), peroxynitrite (ONOO-) and Prostaglandin E₂(PGE₂) were measured in the H₂O₂-Treated renal epithelial cells(LLC-PK1 cell) of mouse. And the effects of Prunellae Spica on the expression of NF-κB (p50, p65), IKK-α, phospho-IκB-α and inflammation-related proteins, COX-2, iNOS, IL-1β and VCAM-1, were examined by western blot. The fluorescent probes, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihyldrorhodamine 123 (DHR 123) were used to estimate the scavenging effect of Prunellae Spica on ․O₂-, NO, ONOO-. Western blot was conducted to assess the protein expression levels of NF-κB (p50, p65), IKK-α, phospho-IκB-α, inflammation-related proteins, COX-2, iNOS, IL-1β, VCAM-1. PH inhibited H₂O₂-treated cell death dose-dependently. It reduced the generation of ·O₂-, NO, ONOO- and PGE₂ in the H₂O₂-treated renal epitheial cells(LLC-PK₁ cell) of mouse in vitro. PH reduced the expression of NF-κB, IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes through means of decreasing activation of NF-κB signaling as well. According to these results, PH has an antioxidative activity and anti-inflammatory effect by regulating the NF-κB pathway. This suggest that PH is expected to be used to regulating inflammatory process and treating inflammation-related disease.

• The Journal of Internal Korean Medicine
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• v.31 no.1
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• pp.153-165
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• 2010

Objectives : The aims of this study were to investigate the cytoprotective, antioxidative and inflammation genes inhibitory effects of Patriniae Radix on the mouse LLC-$PK_1$ cells (renal epithelial cells). Methods : The cytoprotective effect of Patriniae Radix was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of superoxide anion radical (${\cdot}{O_2}^-$) by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), nitric oxide (NO) by 4,5-diaminofluorescein (DAF-2), peroxynitrite ($ONOO^-$) by dihyldrorhodamine 123 (DHR 123) and prostaglandin $E_2$ ($PGE_2$) by $PGE_2$ immunoassay on $H_2O_2$-treated LLC-$PK_1$ cells. For measuring of inflammation genes inhibitory effects, western blot was performed to detect IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 protein level in cytosol fractions from LLC-$PK_1$ cells. Results : Patriniae Radix extract reduced the $H_2O_2$-induced cell death and inhibited the amount of $H_2O_2$-induced ${\cdot}{O_2}^-$, NO, $ONOO^-$, $PGE_2$ generation dose-dependently on the mouse LLC-$PK_1$ cells in vitro. Also Patriniae Radix extract inhibited the expression of IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, COX-2, iNOS, IL-$1\beta$ and VCAM-1 genes dose-dependently by means of decreasing activation of NF-${\kappa}B$. Conclusions : According to above results, it was identified that Patriniae Radix had the cytoprotective, antioxidative and inflammation genes inhibitory effects. So it was suggested that Patriniae Radix would be effective to the treatment for the inflammatory process and inflammation-related diseases.

• The Journal of the Acoustical Society of Korea
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• v.42 no.1
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• pp.50-56
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• 2023

The hydroxyl radical (·OH) and superoxide anion radical (·O₂^- ) generated by the shock wave generated during ultrasonic cavitation collapse in TiO₂ suspension are highly useful because they can sterilize and disinfect. For practical use as a sterilization method without any chemicals, in this study, we proposed a method for evaluating the generation of radicals generated by high-intensity ultrasound emitted to titanium dioxide suspension. In the proposed method, the sonoluminescence phenomenon, which emits light by ultrasonic cavitation decay energy, was utilized, and the degree of radical generation was evaluated through the amount of light energy by sonoluminescence. As a result, even at a low concentration of titanium dioxide of 0.02 wt%, light energy 5 times higher than in the absence of titanium dioxide was received. After that, as the concentration increased by 0.1 wt%, the luminous intensity of sonoluminescence increased linearly by about 14.8 × 10^-12 lm. Therefore, it was confirmed that the radicals generated by radiating high-intensity ultrasound to the titanium dioxide suspension increased linearly as the concentration of titanium dioxide increased within a given concentration range.

• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1096-1101
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• 2008

Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.