• Journal of Life Science
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• v.23 no.1
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• pp.31-37
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• 2013

The present study aimed to investigate effects of ethanol extracts from Hydropsyche kozhantschikovi on cell and DNA damage caused by oxidative stress. In a radical scavenging assay, compared with ascorbic acid used as a control, the level of DPPH (1,1-diphenyl-2-picrylhydrazyl) and that of hydroxyl radicals in H. kozhantschikovi extracts were 60.0% and 43.7%, respectively. The ferrous iron chelating level was 37.5% compared to the chelating value of EDTA (ethylenediaminetetraacetic acid) as a positive control at the same concentration. To verify inhibitory effects of oxidative cell damage induced by reactive oxygen species (ROS), the relative level of lipid peroxidation and the expression level of the p21 protein were compared in extracts-treated and untreated groups. Lipid peroxidation was completely inhibited in the extracts-treated group compared with the radical-only treated group. The level of p21 protein expression was restored to 92.2% of p21 protein expression in the control sample. In addition, DNA cleavage inhibition in the H. kozhantschikovi extracts was 74.1% compared with that of the control group, suggesting that H. kozhantschikovi extracts repress DNA cleavage induced by ROS. Moreover, the phosphorylation ratio of the H2AX protein was 16.7% in the radical-treated group, indicating that the ethanol extracts inhibited 83.3% of DNA damage. Our findings suggest that ethanol extracts from H. kozhantschikovi are effective not only in repressing the oxidation of free radicals and highly toxic hydroxyl radicals, but also in decreasing cell and DNA damage caused by oxidative stress.

• Journal of Life Science
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• v.27 no.7
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• pp.783-789
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• 2017

Malus melliana (Hand.-Mazz.) Rehder (M. melliana) is a Chinese plant that belongs to the Rosaceae family. There have been no previous reports regarding its bioactivity. In this study, the anti-oxidative and anti-inflammatory activities of M. melliana ethanol extract (MMEE) were evaluated using a 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity assay, reactive oxygen species (ROS) scavenging activity assay, nitric oxide (NO) inhibitory activity assay, and the analysis of related protein expressions through Western blot hybridization. MMEE showed potent scavenging activity against DPPH, similar to ascorbic acid, a well-known anti-oxidative agent, which was used as a positive control. MMEE also inhibited hydrogen peroxide-induced ROS in RAW 264.7 cells. Moreover, MMEE induced the expression of an anti-oxidative enzyme, heme oxygenase 1, and its upstream transcription factor, nuclear factor E2-related factor-2, in a dose-dependent manner. On the other hand, MMEE was associated with a reduction in NO production, which was induced by the lipopolysaccharide treatment of RAW 264.7 cells. The expression of inducible nitric oxide synthase, which is the upstream regulator of NO production, was also inhibited. Taken together, these results suggest that MMEE has anti-oxidative and anti-inflammatory properties, thus appearing to be a potential anti-oxidant and anti-inflammatory agent. The further identification of active compounds that confer the biological activities of MMEE may be necessary.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.7
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• pp.801-808
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• 2017

The present study investigated the protective effects of Bifidobacterium bifidum culture supernatants (BbSC) and intracellular cell-free extracts (BbICFE) on human dermal fibroblasts (HDFs) against ultraviolet-B (UV-B) irradiation. HDFs were treated with UV-B, UV-B+BbCS, and UV-B+BbICFE. Treatment of UV-B-irradiated HDFs with BbCS and BbICFE significantly increased cell viability compared to UV-B-irradiated HDFs. BbCS treatment reduced senescence in HDFs by approximately 40.0%. Moreover, sub-G1 phase was significantly reduced in BbCS- and BbICFE-treated HDFs (3.3% and 4.5%, respectively). The effect of UV-B on oxidative damage of HDFs was measured by dichlorofluorescin diacetate. Fluorescence intensity significantly increased in UV-B-irradiated HDFs. Inhibition of cellular reactive oxygen species in HDFs treated with 0.01% BbCS was the highest at 34.1%. Levels of p21 and p53 protein expression induced by UV-B irradiation were reduced by treatment with BbCS and BbICFE (47.0% and 35.6%, respectively). These results show that BbCS and BbICFE reduce UV-B-induced cellular senescence and apoptosis in HDFs. Thus, BbCS and BbICFE can be used as potential agents for protection of UV-B-induced skin cell damage.

• Journal of Korean Medicine Rehabilitation
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• v.25 no.2
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• pp.15-35
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• 2015

Objectives This study was performed to investigate the effects of Bujasasim-tang ethanol extract (BST) on oxidative stress, inflammation and osteoarthritic rat model. Methods To ensure safety of BST, heavy metal levels were measured and cytotoxicity test was done. In vitro, To evaluate antioxidative effects of BST, total phenolic contents, 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, reactive oxygen species (ROS) levels were measured. Also, to evaluate anti-inflammatory effects of BST treated group, total nitric oxide (NO) and pro-inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$) levels were measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In vivo, We injected MIA $50{\mu}l$ (60 mg/ml) into knee joints of rats to induce osteoarthritis. Rats were divided into total 3 groups (normal, control, BST treated group, each n=7). Normal group was not treated at all without inducing osteoarthritis and taken normal diet. Control group was induced osteoarthritis by MIA and taken with 2 ml of distilled water once a day for 4 weeks. BST treated group was induced osteoarthritis by MIA and taken BST 2 ml (200 mg/kg/mouse) once a day for 4 weeks. We evaluated dynamic weight bearing with the Incapacitance Test Meter. At the end of experiment, the rats were sacrificed to observe the functions of liver and kidney, changes of WBC, neutrophil, lymphocyte, monocyte levels in blood, to evaluate the levels of pro-inflammatory cytokines, tissue inhibitor of metallopreteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), prostaglandin $E_2$ ($PGE_2$), leukotriene $B_4$ ($LTB_4$) within serum. We observed change of articular structures by Hematoxylin & Eosin (H&E), safranin-O staining method and measured amount of cartilage by micro CT-arthrography. Statistical analysis was done by unpaired student's t-test with significance level at p<0.05 in SPSS 11.0 for windows. Results 1. Safety of the BST was identified. 2. AST, ALT, BUN, creatinine levels of BST treated group were within normal limit. In vitro, 1. DPPH and ABTS free radical scavenging activities of BST showed dose-dependent increase. 2. ROS production were significantly decreased. 3. Total nitric oxide (NO) and IL-$1{\beta}$ production were decreased. 4. IL-6 and TNF-${\alpha}$ production were significantly decreased. In vivo, 1. Weight bearing ability was significantly increased. 2. WBC, neutrophil, lymphocyte, monocyte levels in blood were decreased. 3. IL-$1{\beta}$ and TNF-${\alpha}$ levels in serum were significantly decreased. and the IL-6 level was decreased. 4. TIMP-1, MMP-9, $LTB_4$, $PGE_2$ levels in serum were significantly decreased. 5. Cartilage volume of BST treated group was significantly increased. Also changes of cartilage, synovial membrane, fibrous tissue were suppressed. Conclusions The results obtained in this study Bujasasim-tang have effects of antioxidative, anti-inflammatory, relieve pain and protection of cartilage. Therefore we expect that Bujasasim-tang is effective treatment for osteoarthritis.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.4
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• pp.193-198
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• 2016

Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzyme defect. G6PD plays a key role in the pentose phosphate pathway, which is a major source of nicotinamide adenine dinucleotide phosphate (NADPH). NADPH provides the reducing equivalents for oxidation-reduction reductions involved in protecting against the toxicity of reactive oxygen species such as $H_2O_2$. We hypothesized that G6PD deficiency may reduce the amount of NADPH in sperms, thereby inhibiting the detoxification of $H_2O_2$, which could potentially affect their motility and viability, resulting in an increased susceptibility to infertility. Methods: Semen samples were obtained from four males with G6PD deficiency and eight healthy males as a control. In both groups, motile sperms were isolated from the seminal fluid and incubated with 0, 10, 20, 40, 60, 80, and $120{\mu}M$ concentrations of $H_2O_2$. After 1 hour incubation at $37^{\circ}C$, sperms were evaluated for motility and viability. Results: Incubation of sperms with 10 and $20{\mu}M\;H_2O_2$ led to very little decrease in motility and viability, but motility decreased notably in both groups in 40, 60, and $80{\mu}M\;H_2O_2$, and viability decreased in both groups in 40, 60, 80, and $120{\mu}M\;H_2O_2$. However, no statistically significant differences were found between the G6PD-deficient group and controls. Conclusion: G6PD deficiency does not increase the susceptibility of sperm to oxidative stress induced by $H_2O_2$, and the reducing equivalents necessary for protection against $H_2O_2$ are most likely produced by other pathways. Therefore, G6PD deficiency cannot be considered as major risk factor for male infertility.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.3 s.52
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• pp.265-271
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• 2005

This study is performed to investigate the effects of citrus essential oils on melanin production in B16 melanoma cells and reactive oxygen species (ROS) generation in RBL 2H3 cells. Five kinds of citrus essential oil (bergamot, grapefruit, lemmon, mandarin, petigrain) did not have any influence on DPPH radical scavenger activity, cell growth and cytotoxicity in B16 melanoma cells. In purified tyrosinase assay, both mandarin and petigrain essential oils dose-dependently inhibited its activity, but bergamot did not. In $1{\mu}M\;{\alpha}-MSH-stimulated$ B16 melanoma cells, all of 5 citrus essential oils inhibited melanin production in $\underline{a}$ dose dependent manner. On the other hand, four kinds of citrus essential oil dose-dependently increased ROS generation in RBL 2H3 mast cells, but mandarin did not. From the above results, it is possible that citrus essential oils nay be developed to be anti-melanogenic agent on the basis of their inhibitory effect on MSH-induced melanin production. Hut we can not rule out the possibility of the induction of allergy and inflammation since citrus essential oils caused ROS generation in RBL 2H3 mast cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.32 no.2 s.57
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• pp.75-79
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• 2006

In this study. we evaluated anti-aging activity of medical plants that protect the skin cell damage induced by UV irradiation. We have investigated diverse biological activities of Selaginella tamariscina as an anti-aging ingredient of cosmetics. S. tamariscina was found to show scavenging activities of radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $65.1{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $40.9 {\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. For testing intracellular ROS scavenging activity, the cultured human dermal fibroblasts were analyzed by increase in dichlorofluorescein (DCF) fluorescence upon exposure to UVB $20 mJ/cm^2$ after treatment of S. tamariscina. UVA-induced MMP-1 protein and mRNA expression in human dermal fibroblasts were reduced in a dose-dependent manner by S. tamariscina. Moreover, S. tamariscina inhibited MMP-2 (gelatinase) activity in UVA-irradiated human dermal fibroblasts assayed by zymography and semi-quantitative RT-PCR. Taken together, these results suggest that S. tamariscina may act as an anti-aging agent by Increasing collagen and preventing the skin cell damage induced by UV irradiation, and imply that S. tamariscina nay be useful as a new ingredient for anti-aging cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.2
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• pp.93-102
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• 2017

In this study, various plant extracts were screened for the effective and safe skin care ingredient according to the antioxidant activity assay guidelines. We selected an optimized combination herbal extract, Charmzone extract (CZE), which composed of Nelumbo nucifera leaves, Saururus chinensis and Orostachys japonica. CZE exerted free radical scavenging activity. It reduced reactive oxygen species formation and increased total antioxidant capacities in human keratinocyte (HaCaT) and normal human dermal fibroblasts (NHDF). CZE also leaded procollagen type 1 secretion in NHDF and decreased cellular melanin contents in B16F10. The production of nitric oxide was decreased by CZE in lipopolysaccharide-stimulated RAW264.7 in a dose dependent manner. Therefore, it is concluded that CZE can be applied for naturally derived anti-aging functional skin care ingredient for anti-oxidation, wrinkle enhancement, whitening, anti-inflammation and wound healing of skin.

• Korean Journal of Plant Resources
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• v.31 no.2
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• pp.102-108
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• 2018

Rubiae radix is root of Runia akane Nakai, it has been used to hemostasis and blood stasis in Korean and China. This study investigated that anti-oxidant and anti-colorectal cancer effect of ERA (ethanol extract of Rubiae radix) and WRA (water extract of Rubiae radix) using RAW 264.7 (murine macrophage from blood) and HCT-116 cells (human colorectal cancer cell line). ERA contained polyphenol ($45.77{\pm}2.03mg/g$) and flavonoid ($22.82{\pm}1.33mg/g$). $500{\mu}M$ $H_2O_2$-induced ROS generation was diminished by $500{\mu}g/m{\ell}$ ERA treatment in RAW 264.7 cells, but not WRA (125, 250, and $500{\mu}g/m{\ell}$). Moreover, caspase-3 activity and DNA fragmentation increased by $500{\mu}g/m{\ell}$ ERA treatment during apoptotic cell death in HCT-116. Results demonstrated that anti-cancer effect of ERA against human colorectal cancer cells is mediated apoptotic cell death and DNA fragmentation through caspase-3 activation. However, further study is required to what active ingredient of ERA are important for anti-oxidant and anti-colorectal cancer effect in vivo.

• Journal of dental hygiene science
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• v.13 no.3
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• pp.321-329
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• 2013

Although it has been reported that lysyl oxidase (LOX) is involved in odontoblastic differentiation, the role of LOX on odontoblastic differentiation by hydrogen peroxide ($H_2O_2$) have not been clarified. In the present study, we investigated whether $H_2O_2$, reactive oxygen species (ROS), is modulated the messenger RNA (mRNA) expression and activity of LOX during odontoblastic differentiation of human dental pulp (HDP) cells. The mRNA expression was quantified by reverse transcriptase polymerase chain reaction (RT-PCR) analysis, and LOX enzyme activity was measured by high sensitive fluorescent assay. Expression of the odontoblastic differentiation marker genes were assessed in the presence and absence of specific small interfering RNAs (siRNAs) of the LOX and LOXL. The $H_2O_2$-induced mRNA expression of LOX family was significant reduction of LOX, LOXL, and LOXL3 mRNA levels in HDP cells. LOX enzyme activity was increased at $H_2O_2$ 0.3 mM for 24 hours. The mRNA expression of alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) was inhibited by LOX- and LOXL-specific siRNAs whereas the mRNA expression of dentin matrix protein1 (DMP1), and dentin sialophosphoprotein (DSPP) was inhibited by LOX-specific siRNA. In LOX enzyme activity, siRNA-induced knockdown of both LOX and LOXL inhibited the total amine oxidase activity in HDP cells, as in the case of mRNA expression. In conclusion, the essential role of $H_2O_2$ on odontoblastic differentiation suggests that its regulation by LOX may have pharmacologic importance in HDP cells.