• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.1 s.49
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• pp.121-125
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• 2005

Most of the cells, specially in the skin, free radicals and reactive oxygen species induced aging, accompanying with diseases. The pinaceae family is very familiar plant with Korean spirit. However, those plants were not estimated for natural products in manufactural process. We employed 4 different plants, Pinus koraienis S et Z. Pinus banksiana LAMBERT. Pinus rigida MILL. Pinus densiflora S. et Z. in pinceae family. The ethanol extracts were prepared for three different parts of plant, wood, bark and leaf, respectively. In this study, we carried out free radical and superoxide radical scavenging assay to investigate the anti-oxidative activity The bark and leaf part of plants showed similar anti-oxidant effect. L-ascorbic acid and $\alpha$-tocopherol at $10{\mu}g/mL$. Also we tried to investigate inhibitory effects of elastase activity in in vitro experiments on pinaceae plant extract as a matters for functional cosmetics. Among those plants, the bark of Pinus rigida MILL. and Pinus densiflora S. et Z. showed inhibitory effect. The cell viability was evaluated with MTT assay. The potential relationship was shown between the cell viability and anti-oxidant effect because the anti-oxidant effects were positively correlated wvith the cell growth in MTT assay. As the results in our experiments, we expect the potential activities of pinaceae as a material of functional cosmetics.

• Nutrition Research and Practice
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• v.17 no.4
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• pp.670-681
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• 2023

BACKGROUND/OBJECTIVES: Oxidative stress is caused by reactive oxygen species and free radicals that accelerate inflammatory responses and exacerbate fatigue. Tormentic acid (TA) has antioxidant and anti-inflammatory properties. Thus, the aim of present study is to determine the fatigue-regulatory effects of TA in H₂O₂-stimulated myoblast cell line, C2C12 cells and treadmill stress test (TST) and forced swimming test (FST) animal models. MATERIALS/METHODS: In the in vitro study, C2C12 cells were pretreated with TA before stimulation with H₂O₂. Then, malondialdehyde (MDA), lactate dehydrogenase (LDH), creatine kinase (CK) activity, tumor necrosis factor (TNF)-α, interleukin (IL)-6, superoxide dismutase (SOD), catalase (CAT), glycogen, and cell viability were analyzed. In the in vivo study, the ICR male mice were administered TA or distilled water orally daily for 28 days. FST and TST were then performed on the last day. In addition, biochemical analysis of the serum, muscle, and liver was performed. RESULTS: TA dose-dependently alleviated the levels of MDA, LDH, CK activity, TNF-α, and IL-6 in H₂O₂-stimulated C2C12 cells without affecting the cytotoxicity. TA increased the SOD and CAT activities and the glycogen levels in H₂O₂-stimulated C2C12 cells. In TST and FST animal models, TA decreased the FST immobility time significantly while increasing the TST exhaustion time without weight fluctuations. The in vivo studies showed that the levels of SOD, CAT, citrate synthase, glycogen, and free fatty acid were increased by TA administration, whereas TA significantly reduced the levels of glucose, MDA, LDH, lactate, CK, inflammatory cytokines, alanine transaminase, aspartate transaminase, blood urea nitrogen, and cortisol compared to the control group. CONCLUSIONS: TA improves fatigue by modulating oxidative stress and energy metabolism in C2C12 cells and animal models. Therefore, we suggest that TA can be a powerful substance in healthy functional foods and therapeutics to improve fatigue.

• Journal of Applied Biological Chemistry
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• v.57 no.3
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• pp.205-210
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• 2014

Persicaria orientalis (L.) Spach (Po) and Potentilla fragarioides var. major Maxim (Pf) extracts were analyzed to investigate anti-inflammation through their suppressing effects on free radicals such as reactive oxygen species (ROS). In addition, with regard to Po and Pf, an analysis was conducted of their inhibitory effect on nitric oxide, which is produced in lipopolysaccharide (LPS)-treated murine macrophage RAW 264.7 cells, and their inhibitory effect on the translocation of the nucleus of nuclear factor-kappa B (NF-${\kappa}B$). The $IC_{50}$ value of ROS, which was induced by $50{\mu}M$ 3-morpholinosydnonimine hydrochloride (SIN-1), was found to be $23.35{\pm}1.27{\mu}g/mL$ due to the effect of the Po extract, and $8.46{\pm}1.22{\mu}g/mL$ due to the effect of the Pf extract. In addition, the $IC_{50}$ value of peroxynitrite treated with the Po extract was $2.19{\pm}0.04{\mu}g/mL$, whereas that of peroxynitrite treated with the Pf extract was $0.80{\pm}0.02{\mu}g/mL$. ROS and peroxynitrite were induced by $50{\mu}M$ 3-morpholinosydnonimine hydrochloride. There was an increase in the amount of nitric oxide in the RAW 264.7 cells treated with LPS ($1{\mu}g/mL$), whereas the level of NO was observed to significantly and dose-dependently decrease in the cells treated with Po and Pf. The amount of nitric oxide produced by the group treated with $10{\mu}g/mL$ of the Pf extract was $11.45{\pm}0.57{\mu}M$. Furthermore, the Po extracts inhibited the translocation of the nucleus of NF-${\kappa}B$ in LPS-treated RAW 264.7 cells. Therefore, it is highly possible that Po and Pf have anti-inflammatory properties.

• Tuberculosis and Respiratory Diseases
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• v.41 no.3
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• pp.215-221
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• 1994

Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.63-71
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• 2004

The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmental facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Juniperus chinensis xylem extract on the UV and SLS-induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. We found that Juniperus chinensis xylem extracts had potent radical scavenging effect by 98％ at 100 $\mu\textrm{g}$/mL. Fluorometric assays of the proteolytic activities of matrix metalloproteinase-l(MMP-1, collagenase) were performed using fluorescent collagen substrates. UV A induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97％ at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79％ at 25 $\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. In this test Juniperus chinensis decreased expression of interleukin 6 about 30％. Expression of prostaglandin E$_2$, (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay (EIA) using PGE$_2$ monoclonal antibody. At the concentrations of 5-50 $\mu\textrm{g}$/mL of the extracts, the production of PGE$_2$ by HaCaT keratinocytes (24 hours after 10 mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p〈0.05). The viability of cultured HaCaT keratinocytes was significantly reduced at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB irradiation, but the presence of these extracts improved cell viability comparing to control after UVB irradiation. We also investigated the protective effect of this extract in sodium lauryl sulfate (SLS)-induced irritant skin reactions from 24 hour exposure. Twice a day application of the extract for reducing local inflammation in human skin was done. Irritant reactions were assessed by various aspects of skin condition, that is, erythema (skin color reflectance) and transepidermal water loss (TEWL). After 5 days the extract was found to reduce SLS-induced skin erythema and improve barrier regeneration when compared to untreated symmetrical test site. In conclusion, our results suggest that Juniperus chinensis can be effectively used for the prevention of UV and SLS-induced adverse skin reactions such as radical production, inflammation and skin cell damage.

• Journal of Life Science
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• v.24 no.3
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• pp.274-283
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• 2014

Oxidative stress causes tissue damage and facilitates the progression of metabolic diseases, including diabetes, cardiovascular heart diseases, and obesity. Lipid accumulation and obesity-related complications have been observed in the presence of extensive oxidative stress. As part of an ongoing study to develop therapeutic supplements, Sargassum sp. were tested for their ability to scavenge free radicals and intracellular reactive oxygen species (ROS), as well as to suppress lipid accumulation. Three species, S. hemiphyllum, S. thunbergii, and Sargassum horneri, were shown to scavenge free radicals in a di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH) assay. In addition, Sargassum sp. was shown to scavenge intracellular ROS and to decrease nitric oxide (NO) production in $H_2O_2$ and lipopolysaccharide (LPS)-induced in RAW264.7 mouse macrophages, respectively. Taken together, the results suggest that Sargassum sp. possess huge potential to relieve oxidative stress and related complications, as well as lipid-induced oxidation. They indicate that S. hemiphyllum, S. thunbergii, and S. horneri are potent functional supplements that can produce beneficial health effects through antioxidant and antiobesity activities, with S. hemiphyllum being the most potent among the Sargassum sp. tested. A potential mechanism for the effect of Sargassum sp. on the suppression of lipid accumulation in differentiating 3T3-L1 mouse preadipocytes through deactivation of the peroxisome proliferator-activated receptor ${\gamma}$ (PPAR ${\gamma}$) is presented.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.1
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• pp.11-20
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• 2014

In this work, the antioxidative effects and active component analysis of Gnaphalium affine D. DON. (G. affine) extracts were investigated. All experiments were performed with 70% ethanol extract, ethyl acetate fraction and aglycone fraction of the G. affine extract. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of ethyl acetate fraction ($6.15{\mu}g/mL$) of the G. affine was higher than that of (+)-${\alpha}$-tocopherol ($8.89{\mu}g/mL$), which is known as a reference control. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of the 70% ethanol extract ($1.60{\mu}g/mL$), ethyl acetate fraction ($0.075{\mu}g/mL$) and aglycone fraction ($2.28{\mu}g/mL$) of extract of G. affine on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system using the luminol-dependent chemiluminescence assay were much higher than that of L-ascorbic acid ($6.88{\mu}g/mL$). The cellular protective effects of 70% ethanol extract (${\tau}_{50}$ = 52.0 min) and aglycone fraction of the extract (${\tau}_{50}$ = 60.6 min) on the $^1O_2$-induced cellular damage of human erythrocytes were exhibited the higher protective effect than (+)-${\alpha}$-tocopherol (${\tau}_{50}$ = 38.0 min), known as a lipophilic antioxidant. TLC and HPLC were used to analyse active components in the aglycone fraction of the extract. Results showed that luteolin and apigenin were main components. These results suggest that the G. affine extract can be applied to an effective antioxidant in scavenging ROS including radicals.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.2
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• pp.139-147
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• 2005

The efficacy of extraction from Inonotus obliquus was examined from the points of antioxidative characteristics and some antioxidative compounds. To enhance the efficient extraction for the effective components from Inonotus obliquus, temperature-stepwise water extraction method was applied. Temperature-stepwise water extracts were prepared for 8 hrs as follows: the first extract at 8$0^{\circ}C$, the second extract from the residue of the first extract at 10$0^{\circ}C$, and the third extract from the residue of the second extract at 12$0^{\circ}C$. Antioxidativeactivities were determined by electron-donating ability of DPPR - free radical, scavenging ability of ABTS$.$$^{+}$radical cation, and by inhibiting ability of linoleic acid autoxidation. In results, the first extract showed the least antioxidant capacity, and the third extract showed the highest antioxidant capacity. The third extract also had the greatest amounts of phenolic compounds and flavonoids. Amounts of phenolic compound from each extract were almost proportional to the radical scavenging activities and linoleic acid autoxidation inhibiting ability (r=0.960∼0.980, regression analysis). Furthermore, the effect of the pooled extract of all three extractions of Inonotus obliquus on the lipid peroxidation reacted with active oxygen species (KO$_2$, $H_2O$$_2$, $.$OH) and metals (Fe$^{2+}$, CU$^{2+}$) was evaluated by measuring the formation of thiobarbituric acid reactive substances (TBARS). The pooled Inonotus obliquus extracts lowered the amounts of TBARS formed by all of the active oxygen species and metals. Especially, these lowering effects were pronounced in the reaction with $.$OH and Fe$^{2+}$. These results suggest that the pooled temperature-stepwise extract from Inonotus obliquus could be potential functional materials to reduce the oxidation of lipids and other compounds induced by free radicals.adicals.

• KSBB Journal
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• v.10 no.5
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• pp.561-567
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• 1995

Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxygen toxicity induced by paraquat was studied. In aerobic culture condition, yeasts lacking MnSOD (milochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared with wild type were observed under anaerobic condition. When exposed to paraquat, the growth of yeasts lacking CuZnSOD was severely affected by higher than 0.01mM of paraquat in culture medium. By the analysis of several cellular components ivolved in free radical generating and scavenging system, it was found that, under aerobic condition, the content of lipid peroxides in cell membrane as well as cellular activity of glutathion peroxidase of CuZnSOD-deficient mutants was increased in the presence of paraquat, although significant decrease of catalase activity was observed in those stratns. In MnSOD-deficient yeast, however, increment in cellular activity of glutathion peroxldase and catalase by paraquat was observed without any deterioration of membrane lipid. It implies that the lack of mitochondrial SOD could be compensated by both of glutathion peroxldase and catalase, but that only glutathion peroxidase might act for CuZnSOD in cytoplasm. In contrast, all of SOD-deficient mutants showed a significant decrease in catalase activity, but slight increase in the activities of glutathion peroxidase, when cultivated anaerobically in the medium containing paraquat. Nevertheless, any significant changes of lipid peroxides in cell membranes were not observed during anaerobic cultivation of SOD-deficient mutants. It suggests that a little amount of free radicals generated by paraquat under anaerobic condition could be sufficiently overcome by glutathion peroxidase but not by catalase.

• Journal of Life Science
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• v.31 no.6
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• pp.559-567
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• 2021

This study compared the nutritional characteristics and antioxidant effects of sea mustards sourced from five different areas (Barammaegi, Gultongmeori, Chanmulgae, Johongtaek, and Goraedeung) in Taejongdae, Youngdo, Busan. The contents of total flavonoids and phenols and fatty acid composition were measured. To evaluate their antioxidant effects, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays were used. Acetone/methylene chloride (A+M) extracts from all the sea mustards contained higher amounts of total flavonoids and phenols than methanol (MeOH) extracts. Among the sea mustards obtained from the different areas, the total flavonoid and total phenolic content of the A+M extract of the sea mustard from Gultongmeori was 1.44±0.04 mg/g and 1.72±0.06 mg/g, respectively. In terms of the fatty acid composition, the Gultongmeori sea mustard had higher percentages of total n-6, total n-3, eicosapentaenoic acid (EPA, 20:5n-3), and docosahexaenoic acid (DHA, 22:6n-3) than the sea mustards from the other areas. The A+M extract of the sea mustard from Gultongmeori was more effective in terms of scavenging free radicals as compared with that of the other sea mustards, as assessed by the DPPH and ABTS assays (p<0.05). In a 120-minute reactive oxygen species (ROS) production assay, all the extracts tested decreased cellular ROS production induced by H₂O₂ compared to that produced by exposure to an extract-free control (p<0.05). The extracts from Barammaegi and Gultongmeori had a greater inhibitory effect on cellular ROS production. These results indicated that the antioxidant effects of sea mustards might be associated with a higher amount of flavonoids and phenols. This study suggests that food-processed products from sea mustard can be developed as functional foods for promoting health in the local population.