• International Journal of Stem Cells
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• v.15 no.4
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• pp.359-371
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• 2022

Background and Objectives: The goal of this study was to investigate the mechanism of mesenchymal stem cell (MSC)-derived microRNA (miR)-150-5p-expressing exosomes in promoting skin wound healing through activating PI3K/AKT pathway by PTEN. Methods and Results: Human umbilical cord (HUC)-MSCs were infected with miR-150-5p overexpression and its control lentivirus, and HUC-MSCs-derived exosomes (MSCs-Exos) with stable expression of miR-150-5p were obtained. HaCaT cells were induced by H₂O₂ to establish a cellular model of skin injury, in which the expression of miR-150-5p and PTEN and the phosphorylation of PI3K and AKT were evaluated. HaCaT cells were transfected with pcDNA3.1-PTEN or pcDNA3.1 and then cultured with normal exosomes or exosomes stably expressing miR-150-5p. Cell proliferation was inspected by CCK-8. Cell migration was detected by scratch test and cell apoptosis by flow cytometry. The starBase tool was used to predict the binding site of miR-150-5p to PTEN. Dual-luciferase reporter assay and RIP assay were applied to assess the interaction between miR-150-5p and PTEN. In H₂O₂-induced HaCaT cells, the miR-150-5p expression decreased, and PTEN expression increased in a concentration-dependent manner. MSCs-Exos promoted the growth and migration of H₂O₂-induced HaCaT cells and inhibited their apoptosis. In addition, overexpression of exosomal miR-150-5p enhanced the protective effect of MSCs-Exos on H₂O₂-induced HaCaT cells; PTEN overexpression in HaCaT cells partially restrained miR-150-5p-mediated inhibition on H₂O₂-induced injury in HaCaT cells. PTEN was a target gene of miR-150-5p. MiR-150-5p regulated PI3K/AKT pathway through PTEN. Conclusions: MSCs-derived miR-150-5p-expressing exosomes promote skin wound healing by activating PI3K/AKT pathway through PTEN.

• Journal of Life Science
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• v.19 no.5
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• pp.633-638
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• 2009

We investigated the inhibitory effects of solvent extracts from dried tuna on the growth of cancer cell lines (HT1080 human fibrosarcoma and HT-29 human colon cancer cells) and $H_2O_2$-induced oxidative stress. Inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts on the growth of HT1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was more significant on the growth of HT1080 cells, and A+M extracts had a higher inhibitory effect compared to MeOH extracts. The treatments of hexane, 85% aq. methanol, butanol and water fractions significantly inhibited the growth of both cancer cells (p<0.05). Among the fractions, hexane and 85% aq. methanol fractions showed higher inhibitory effects. In order to determine the protective effect on $H_2O_2$-induced oxidative stress, a DCHF-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including crude extracts of dried tuna, appeared to significantly reduce the levels of intracellular reactive oxygen species (ROS) with dose responses (p<0.05). Among the fractions, BuOH and 85% methanol fractions showed a higher protective effect on the production of lipid peroxides. These results indicate that the consumption of tuna may be recommended as a potent functional food for preventing cellular oxidation and cancer.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.4
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• pp.357-363
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• 2011

In this study, the antioxidative effects and inhibitory activities on tyrosinase of Rheum undulatum (R.undulatum) L. extracts were investigated. 50 % ethanol extract, ethyl acetate and aglycone fractions of R. undulatum L. were used in experiments. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activities ($FSC_{50}$) of R. undulatum L. extracts was lower than (+)-${\alpha}$-tocopherol, known as a typical antioxidant. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of aglycone fraction on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system using the luminol-dependent chemiluminescence assay showed the most prominent effect at a concentration of $0.265\;{\mu}g/mL$. The cellular protective effects of extract/fractions of R. undulatum L. on the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim}50\;{\mu}g/mL$). Especially, aglycone fraction in $10\;{\mu}g/mL$ concentration showed the most protective effect among extracts (${\tau}_{50}$, 757.0 min). The inhibitory effects ($IC_{50}$, $11.20\;{\mu}g/mL$) on tyrosinase of aglycone fraction was much higher than arbutin ($226.88\;{\mu}g/mL$), known as a whitening agent. These results indicate that R. undulatum L. extracts can be used as antioxidant. Particularly, aglycone fraction of R. undulatum L. showed superior antioxdative activity and high inhibitory effect on tyrosinase. Therefore, aglycone fraction of R. undulatum L. could be applicable to new functional cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.3
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• pp.275-282
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• 2012

In this study, the antioxidative effects of extracts from different parts of Juncus effusus L. were investigated. The three parts (above-ground part, below-ground part, medulla part) were selected. 50 % ethanol extract, ethyl acetate and aglycone fractions of J. effusus L. were used in experiments. The highest DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activities ($FSC_{50}$) was shown by medulla part (42.9 ${\mu}g/mL$) in 50 % ethanol extracts, below-ground part (12.1 ${\mu}g/mL$) in ethyl acetate fractions, and below-ground part (12.1 ${\mu}g/mL$) in aglycone fractions. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system using the luminol-dependent chemiluminescence assay showed the most prominent effect of medulla part (0.29 ${\mu}g/mL$) in 50 % ethanol extracts, below-ground part (0.25 ${\mu}g/mL$) in ethyl acetate fractions, and medulla part (0.20 ${\mu}g/mL$) in aglycone fractions. The cellular protective effects of extract/fractions of J. effusus L. on the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner (0.5 ~ 10 ${\mu}g/mL$). Especially, aglycone fraction of medulla part at a concentration of 10 ${\mu}g/mL$ showed the most prominent protective effect among all extracts (${\tau}_{50}$, 321.0 min). These results indicate that extracts from below-ground part and medulla part of J. effusus L. extracts can be used as an natural antioxidant. Particularly, J. effusus L. can protect suggesting a high ${\tau}_{50}$ skin where many $^1O_2$ was generated by sunlight exposure.

• Applied Chemistry for Engineering
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• v.26 no.1
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• pp.29-34
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• 2015

N,N'-Diferuloyl-putrescine (DFP) present in plants such as Sophora japonica has been reported to have skin depigmentative and antioxidative activities. In this study, DFP, usually presents in nature a very little amount and its derivative (DFP-D) were synthesized in a large quantity for the use as functional cosmetical materials. The antioxidative activities of synthesized DFP and DFP-D were evaluated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, chemiluminescence assay, and cell protective effect induced by $^1O_2$, stress. DFP and DFP-D showed DPPH radical scavenging activities ($FSC_{50}$) at $61.25{\pm}2.25{\mu}M$ and $12.92{\pm}0.72{\mu}M$, respectively. ROS (reactive oxygen species) scavenging activities ($OSC_{50}$) in the $Fe^{3+}-EDTA/H_2O_2$ system of DFP and DFP-D were 2 times ($1.84{\pm}0.12{\mu}M$) and 13 times ($0.174{\pm}0.01{\mu}M$), respectively higher than that of L-ascorbic acid. $^1O_2$, one of ROS playing a key role in the skin photo-aging, induces cellular membrane damages. DFP-D ($50{\mu}M$) showed good cell protective effects (${\tau}_{50}=80.2min$) about 2 times more than that of (+)-${\alpha}$-tocopherol (${\tau}_{50}=43.6min$). These results suggest that the great antioxidative activities of DFP and DFP-D could be applied to cosmetic industries as functional cosmetic materials.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.4
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• pp.297-304
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• 2012

In this study, the antioxidative effects and inhibitory activities on tyrosinase of Lindera obtusiloba Blume (L. obtusiloba Blume) extracts were investigated. 50 % ethanol extract, ethyl acetate and aglycone fractions of L. obtusiloba Blume were used in experiments. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activities of ethyl acetate fraction of L. obtusiloba Blume was higher than (+)-${\alpha}$-tocopherol, known as a typical antioxidant. Reactive oxygen species (ROS) scavenging activities of extract and fractions of L. obtusiloba Blume on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were similar to L-ascorbic acid, well known as a strong antioxidant. The cellular protective effects of 50 % ethanol extract and ethyl acetate of L. obtusiloba Blume on the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim}25{\mu}g/mL$). Ethyl acetate fraction in $10{\mu}g/mL$ concentration showed the most protective effect among extracts (${\tau}_{50}$ = 361.0 min). The inhibitory effects on tyrosinase of ethyl acetate and aglycone fractions were higher than arbutin, known as a whitening agent. These results indicate that L. obtusiloba Blume extracts can be used as antioxidant, and could be applicable to functional cosmetic ingredient.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.4
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• pp.259-268
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• 2008

In this study, the antioxidative effects, inhibitory effects on tyrosinase and elastase and components of Castanea crenata leaf were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl radical, DPPH) scavenging activity ($FSC_{50}$) of extract / fractions of Castanea crenata left was in the order: 50% ethanol extract ($13.6{\mu}g/mL$) < ethyl acetate fraction (6.2) < aglycone fraction (2.1). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$ of extract / fractions from Castanea crenata leaf extract / fractions on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was in the order: aglycone fraction (0.8) < 50% ethanol extract (0.5) < ethyl acetate fraction (0.3). The scavenging activity ($IC_{50}$ for ${O_2}^{{\cdot}\;-}$ (superoxide anion radical) generated by NBT method was in the order: ethyl acetate fraction (145.5) < aglycone fraction (65.5). The protective effects on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction exhibited the most prominent cellular protective effect (${\tau}_{50}$, $191.9{\pm}12.2\;min$ at $10{\mu}g/mL$). The inhibitory effect of aglycone fraction ($9.1{\mu}g/mL$) on elastase was higher than oleanolic and ($13.7{\mu}g/mL$). And the inhibitory effect of aglycone fraction ($21.6{\mu}g/mL$) on tyrosinase was higher than arbutin ($226.2{\mu}g/mL$). But 50% ethanol extract rarely exhibited the inhibitory activity on tryosinase and elastase. Flavonoids were contained in Castanea crenata left (96.3 mg / 100 g dried Castanea crenata leaf). And flavonoids contained in ethyl acetate fraction were kaempferol, quercetin, quercitrin, and so on. Quercitrin is the most abundant component. These results indicate that extract / fractions of Castanea crenata can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging free radical and ROS, Castanea crenata leaf extract/ fractions could be used as new cosmeceutical for whitening and anti-wrinkle products.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.109-115
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• 2012

Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

• 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 the Society of Cosmetic Scientists of Korea
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• v.35 no.3
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• pp.209-217
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• 2009

In this study, the antibacterial activity, antioxidative effects, inhibitory effects on tyrosinase of Inula britannica flower extracts were investigated. MIC values of ethyl acetate fraction from Inula britannica flower on P. acnes 0.25 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction was the highest in the P. acnes. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of ethyl acetate fraction of Inula britannica flower was $8.55{\mu}g$/mL. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some fInula britannica flower extracts on ROS generated in $Fe^{3+}$- EDTA/$H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were ethyl acetate fraction $0.24{\mu}g$/mL. Ethyl acetate fraction showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of Inula britannica flower on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Inula britannica flower extracts suppressed photohemolysis in a concentration dependent manner ($5{\sim}100{\mu}g$/mL), particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect ($\tau_{50}$, 164.15 min at $25{\mu}g$/mL). The inhibitory effect of Inula britannica flower extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase of some Inula britannica flower extracts were high. Ethyl acetate fraction has $IC_{50}$ of $87.03{\mu}g$/mL. These results indicate that extract/fractions of Inula britannica flower can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And inhibitory activity on tyrosinase of the ethyl acetate fraction and high potential as bactericide against the skin pathogenic bacteria could be applicable to new functional cosmetics for antioxidant, antiaging, antibacterial activity.