• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.1
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• pp.8-16
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• 2013

We investigated the effect of red cabbage extract (RCE) on cell death in MDA-MB-231 human breast cancer cells. Cells were cultured in the presence 1.0, 1.5, and 2.0 mg/mL concentrations of RCE for 24 hours. MTT assays demonstrated that mitochondrial dehydrogenase activities decreased in a dose-dependent manner in cells (p<0.05). In contrast, the proportion of dual staining with Hoechst 33342/ethidium bromide (EtBr) for cell death increased in a dose-dependent manner in cells (p<0.05). Flow cytometry assays revealed that cell death caused by an apoptotic program increased in a dose-dependent (p<0.05). Also, increased ROS accumulation in cells, as revealed by DCF-DA staining, was observed in a dose-dependent fashion (p<0.05). The apoptosis suppressor gene Bcl-2 decreased significantly at the mRNA level. Pro-apoptotic genes Bax and caspase-3, genes that are related to the last stage of apoptosis significantly increased. The Bcl-2/Bax ratio which is an important indicator of apoptosis, was found to have significantly decreased dose dependence. These results taken together indicate that the effect of red cabbage extract induces cell death in MDA-MB-231 human breast cancer cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.1
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• pp.87-94
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• 2016

Skin damage is mainly caused by environmental factors such as ultraviolet light, heat, and smoking. It is known that reactive oxygen species production is commonly involved in the pathogenesis of skin damage induced by these factors, causing skin aging. Pyrus pyrifolia Nakai continues to be a popular and highly consumed fruit in many countries with known beneficial effects including antitumor, antioxidative, and anti-inflammatory effects. However, there is no evidence of a therapeutic effect of Pyrus pyrifolia extract (PPE) against skin aging via inhibition of mitochondria-mediated apoptosis. In this study, we investigated PPE protective effect against photoaging induced by UVB ($50mJ/cm^2$) in HS68 human dermal fibroblasts. Lactate dehydrogenase assay showed that PPE significantly protected HS68 cells against UVB-induced damage in a dose-dependent manner. Other assays using DCF-DA demonstrated that PPE protected HS68 cells by regulating reactive oxygen species production. PPE also regulated mitochondrial dysfunction and mitochondrial membrane potential induced by UVB, and inhibited UVB-induced caspase-3 activity. These results indicate that PPE protects human dermal fibroblasts from UVB-induced damage by regulating the oxidative defense system.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.46 no.3
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• pp.253-263
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• 2020

Reactive oxygen species (ROS) plays an important role in maintaining homeostasis. However, excessive ROS production damages cellular components such as proteins, lipids, and nucleic acids and promotes skin aging. In this study, we confirmed the antioxidant effect of CSYJE to prevent excessive oxidative stress. First, DPPH and ABTS assays were performed to confirm the antioxidant effect of CSYJE and the radical scavenging activity was confirmed depending on the concentration. As a result of performing the MTT assay to confirm the cell viability, it was confirmed that there was no cytotoxicity at a concentration of 1,000 ㎍/mL. As a result of western blotting to confirm the expression levels of the antioxidant-related proteins nuclear-E2-related factor 2 (Nrf2) and Heme oxygenase-1 (HO-1), it was confirmed that the expression was increased in a concentration-dependent manner. After inducing ROS with lipopolysaccharide (LPS), an intracellular ROS-causing substance, DCF-DA was performed to confirm the inhibitory effect of ROS production, and the inhibition of ROS production was confirmed to concentration-dependent. Real-time RT-PCR was performed to confirm the mRNA expression level of inflammatory cytokines and inflammatory mediator caused by ROS generation, mRNA expression was reduced in a dose dependent manner. Therefore, this study confirmed the antioxidant effect of CSYJE through the Nrf2/HO-1 signaling pathway, which suggests that CSYJE can be used as an antioxidant cosmetic material by inhibiting free radicals.

• The Journal of Internal Korean Medicine
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• v.31 no.1
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• pp.54-65
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• 2010

Objective : The water extract of Chungsimyeonja-eum (CSYJE) has traditionally been used in treatments of heart diseases and brain diseases in Oriental medicine. However, little is known about the mechanism by which CSYJE protects neuronal cells from injury damages. Therefore, in this study we attempted to elucidate the mechanism of the cytoprotective effect of the CSYJE extract on glutamate-induced C6 glial cell death. Methods : Cultured cells were pretreated with CSYJE and exposed to glutamate, cell damage was assessed by using MTT assay and propidium iodide (PI), probe 2',7'-dichlorofluorescein diacetate (DCF-DA) staining. Western blotting was performed using anti-procaspase-3 and anti-PARP, respectively. Result : We determined the elevated cell viability by CSYJE extract on glutamate-induced C6 glial cell death. Glutamate induced DNA fragmentation on C6 glial cells but pre-treatment with CSYJE inhibited DNA fragmentation. One of the main mediators of glutamate-induced cytotoxicity was known to generation of reactive oxygen species (ROS). Pre-treatment with CSYJE inhibited this ROS generation from glutamate-stimulated C6 glial cells. Also, we identified that the ROS-induced DCF-DA green fluorescence was reduced by CSYJE pre-treatment. The critical markers of apoptotic cell death are the cleavages of procaspase-3 protease and PARP proteins, so we checked the expression level and cleavages of procaspase-3 protease and PARP proteins. Glutamate-treated C6 glial cells showed the cleavages of procaspase-3 protease and PARP proteins and followed the reduction of expression of these proteins. Conclusion : These findings indicate that CSYJE may prevent cell death from glutamate-induced C6 glial cell death by inhibiting the ROS generation and procaspase-3 and PARP expression.

• Journal of the Korean Applied Science and Technology
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• v.37 no.1
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• pp.56-65
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• 2020

This study attempted to review the possibility of Psidium guajava leaf extract as a cosmetics ingredient by measuring antioxidant activity through ABTS radical scavenging activity, cytotoxicity in RAW 264.7 macrophages, ROS generation-inhibiting effects through DCF-DA assay and antimicrobial activity, and the results found the followings: The Psidium guajava leaf extract revealed excellent ABTS radical scavenging activity. In RAW 264.7 macrophages, no cytotoxicity was found. The ROS generation in the cells was reduced in a dose-dependent manner. The antimicrobial activities were observed in the following strains: S. aureus, E. coli, C. albicans and P. acnes. In terms of minimum inhibitory concentration (MIC) on each strain, which ranged from 0.25 to 1 mg/mL, C. albican was the lowest, followed by E. coli, S. aureus and P. acnes. The above results confirmed the effects of Psidium guajava leaf extract: antioxidant activity, inhibition of ROS generation in the cells, antimicrobial effects on skin flora which causes inflammation. Therefore, it appears that the extract would be available as a cosmetics ingredient which is free of toxins and side effects.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.48 no.3
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• pp.213-223
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• 2022

In this study, the anti-inflammatory and antioxidant effects of aerial parts of Rumex acetosa L. extract were confirmed to prevent various inflammatory diseases and skin aging caused by excessive oxidative stress. As a result of ABTS assay, it was confirmed that the radical scavenging ability increased in a concentration-dependent manner. ROS inhibitory ability was confirmed through DCF-DA assay, and concentration-dependent inhibition of ROS production was confirmed. The effect of inhibiting cell nuclear damage according to ROS was confirmed through DAPI staining. In addition, it was confirmed that the mRNA expression levels of iNOS and COX-2 were inhibited in a concentration-dependent manner through qPCR. As a result of confirming the protein levels of iNOS and COX-2 by western blotting, iNOS was significantly decreased at all concentrations, and COX-2 was significantly decreased at 800 ㎍/mL. The inhibitory effect on the production of NO generated by iNOS was confirmed by NO assay, and NO was decreased in a concentration-dependent manner. In addition, phosphorylation of ERK and JNK in the MAPKs signaling pathway were inhibited. Therefore, Rumex acetosa L. has the potential to be used as an anti-inflammatory and antioxidant cosmetic raw material by showing anti-inflammatory and antioxidant effects through the MAPKs pathway.

• Journal of the Korean Applied Science and Technology
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• v.39 no.5
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• pp.644-651
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• 2022

The skin is the largest organ of the human body and protects the inside of the body. Ultraviolet rays cause various inflammatory reactions in the skin, including photoaging and oxidative damage. The purpose of this study is to investigate the protective effect of Saponaria extract by irradiating UVB on fibroblasts. In this study, the effectiveness of Saponaria showing protective activity against UVB-induced cytotoxicity, oxidative cell death, and NO and PGE₂ production was evaluated. HS68 cells were irradiated with UVB(120 mJ/cm²) and treated with Saponaria extract at various concentrations of 100, 200, and 400 ㎍/mL for 24 hours. Intracellular reactive oxygen species (ROS) generated by ultraviolet B were detected using a spectrofluorometer after DCF-DA staining. Lipid peroxidation was also analyzed by measuring the level of 8-isoprostane secreted into the culture medium. As a result, treatment with Saponaria extract effectively inhibited UVB-induced cytotoxicity. Oxidative cell damage was mediated by PGE₂ in UVB-induced HS68 fibroblasts, which was significantly inhibited by Saponaria extract treatment. In addition, it was evaluated that the protective effect of these extracts was mediated by the inhibition of intracellular ROS production and lipid peroxidation in a concentration-dependent manner. These results suggest that Saponaria extract can be used as an anti-aging functional material because it inhibits skin damage mediated by oxidative stress caused by UVB and exhibits a cellular protective effect.

• Journal of Life Science
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• v.27 no.7
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• pp.796-804
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• 2017

This study was carried out to evaluate the antioxidant effect of methanolic extract of Houttuynia cordata (HCME) and to identify a compound having antioxidant effect. The ethyl acetate fraction of HCME showed the highest antioxidant effect in organic solvent fractions. The fraction was then separated into 12 fractions by open column chromatography. Among these fractions, the fraction 10 (Fr. 10) with the highest antioxidant activity was isolated, and its antioxidant effect was evaluated by DPPH radical scavenging activity, reducing power, TBARS, cell viability, DNA oxidation and DCF fluorescence. The Fr. 10 at a $64{\mu}g/ml$ showed 60% of inhibitory effect similar to that of vitamin C at $10{\mu}g/ml$, compared with blank group. The Fr. 10 at $64{\mu}g/ml$ showed 264% of reducing power, compared with blank group. TBARS assay showed that the Fr. 10 at $64{\mu}g/ml$ had 35.5% of inhibitory effect similar to that of vitamin E at $1,000{\mu}g/ml$, compared with blank group. The Fr. 10 above $32{\mu}g/ml$ displayed cytotoxicity. However, it was observed that the Fr. 10, above $1{\mu}g/ml$ reduced DNA damage. DCF fluorescence assay showed that the Fr. 10 inhibited oxidative stress by $H_2O_2$ in a dose dependent manner. The compound of Fr. 10 was identified to be rutin whose molecular weight is 610 by the IR and LC-MS analyses. Therefore, these results suggest that the rutin of Fr. 10 could use as a natural antioxidant for development of cosmetics and functional foods.

• The Journal of Internal Korean Medicine
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• v.29 no.4
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• pp.835-845
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• 2008

Objectives : The purpose of this study was to investigate the effect of FS for the modulation of ROS and MMP in a hypoxic model of cultured rat cortical cells. Methods : For the effect of FS on the viability, FS was added to culture media (neurobasal supplemented with B27) and cell viability was measured by LDH assay. To investigate the effects of FS on ROS generation and MMP preservation, cells grown in FS-containing media were given a hypoxic shock(2% $O_2/5%$ $CO_2$, $37^{\circ}C$, 3 hrs) on DIV 10, stained with $H_2DCF-DA$(10 nM) and JC-1, respectively, and observed by fluorescent microscope. Results : 1. FS has a protective effect of cortical cells in both normoxia and hypoxia. 2. FS reduced the generation of ROS and this reduction was especially significant at 3 days after hypoxia. 3. FS was effective for the maintenance of MMP in hypoxia, and this efficacy was especially significant at 3 days after hypoxia. Conclusions : Taken together, these results indicate that FS attenuates ROS generation and MMP dissipation, which eventually protects from neuronal cell death in hypoxia.

• Nutrition Research and Practice
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• v.9 no.6
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• pp.586-591
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• 2015

BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount$^{TM}$ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2',7'-dichlorofluoroscein diacetate ($H_2DCF$-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and $10{\mu}M$) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole ($3{\mu}M$)-treated cells in a dose-dependent manner. Rhamnetin (1 and $3{\mu}M$) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.