• Herbal Formula Science
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• v.31 no.4
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• pp.265-281
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• 2023

Objectives : Ukgan-san plus Citri Pericarpium and Pinelliae Rhizoma (UCP) is used as a traditional herbal formula in Korea and Japan for treatment of fever, fever-induced convulsions, and liver dysfunction and so on. In this study, we investigated the cytoprotective effect and underlying mechanism of UCP against oxidative stress induced by cotreatment of arachidonic acid (AA) and iron. Methods : To evaluate the hepatoprotective effects of UCP against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The changes in intracellular reactive oxygen species (ROS), glutathione (GSH), and mitochondrial membrane permeability (MMP) were investigated against to the oxidative stress. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) and its downstream target genes were examined by immunoblot analysis. Results : Treatment of UCP increased the cell viability and altered the expression levels of apoptosis-related proteins such as PARP, caspase-9, caspase-3, Bcl-2. UCP also inhibited the GSH depletion, excessive ROS production and mitochondrial dysfunction induced by AA + iron. In addition, the Nrf2 and the Nrf2 target genes activation were increased by UCP. Conclusions : These results indicated that UCP has the ability to protect against oxidative stress-induced hepatocyte damage, which may be mediated with Nrf2 pathway.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.311-319
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• 2023

Caveolin-1 (Cav-1) is the main structural component of the caveolae on the plasma membrane, which regulates various cellular processes, including cell growth, differentiation, and endocytosis. Although a recent study demonstrated that Cav-1 might be involved in diabetes-associated inflammation, its exact role in the intestine was unclear. In this study, we examined the intestinal expression of Cav-1 in diabetic conditions. We also investigated its effect on lipopolysaccharide (LPS)-induced inflammation by expressing this protein in human intestinal Caco-2 cells lacking Cav-1. We observed that increased Cav-1 levels and decreased expression of tight junction proteins affected intestinal permeability in high-fat diet-induced diabetic mice. When Caco-2 cells were treated with LPS, Cav-1 enhanced the NF-κB signaling. Moreover, LPS reduced the expression of tight junction proteins while it increased cell-cell permeability and reactive oxygen species generation in Caco-2 cells and this effect was amplified by cav-1 overexpression. LPS treatment promoted phosphorylation of tyrosine-14 (Y14) on Cav-1, and the LPS-induced NF-κB signaling was suppressed in cells expressing non-phosphorylatable Cav-1 (tyrosine-14 to phenylalanine mutant), which reduced intestinal barrier permeability. These results suggest that Cav-1 expression promotes LPS-induced inflammation in Caco-2 cells, and phosphorylation of Y14 on Cav-1 might contribute to the anti-inflammatory response in LPS-induced NF-κB signaling and cell permeability.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.379-386
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• 2024

Basophils and mast cells are specialized effector cells in allergic reactions. Haliotis discus hannai (abalone), is valuable seafood. Abalone male viscera, which has a brownish color and has not been previously reported to show anti-allergic activities, was extracted with acetone. Six different acetone/hexane fractions (0, 10, 20, 30, 40, and 100%) were obtained using a silica column via β-hexosaminidase release inhibitory activity-guided selection in phorbol myristate acetate and a calcium ionophore, A23187 (PMACI)-induced human basophils, KU812F cells. The 40% acetone/hexane fraction (A40) exhibited the strongest inhibition of PMACI-induced-β-hexosaminidase release. This fraction dose-dependently inhibited reactive oxygen species (ROS) production and calcium mobilization without cytotoxicity. Western blot analysis revealed that A40 down-regulated PMACI-induced MAPK (ERK 1/2, p-38, and JNK) phosphorylation, and the NF-κB translocation from the cytosol to membrane. Moreover, A40 inhibited PMACI-induced interleukin (IL)-1β, IL-6, and IL-8 production. Anti-allergic activities of A40 were confirmed based on inhibitory effects on IL-4 and tumor necrosis factor alpha (TNF-α) production in compound (com) 48/80-induced rat basophilic leukemia (RBL)-2H3 cells. A40 inhibited β-hexosaminidase release and cytokine production such as IL-4 and TNF-α produced by com 48/80-stimulated RBL-2H3 cells. Furthermore, it's fraction attenuated the IgE/DNP-induced passive cutaneous anaphylaxis (PCA) reaction in the ears of BALB/c mice. Our results suggest that abalone contains the active fraction, A40 is a potent therapeutic and functional material to treat allergic diseases.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.596-605
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• 2024

Anthocyanins belong to phenolic pigments and are known to have various pharmacological activities. This study aimed to investigate whether anthocyanins could inhibit hydrogen peroxide (H₂O₂)-induced oxidative damage in human retinal pigment epithelial ARPE-19 cells. Our results indicated that anthocyanins suppressed H₂O₂-induced genotoxicity, while inhibiting reactive oxygen species (ROS) production and preserving diminished glutathione. Anthocyanins also suppressed H₂O₂-induced apoptosis by reversing the Bcl-2/Bax ratio and inhibiting caspase-3 activation. Additionally, anthocyanins attenuated the release of cytochrome c into the cytosol, which was achieved by interfering with mitochondrial membrane disruption. Moreover, anthocyanins increased the expression of heme oxygenase-1 (HO-1) as well as its activity, which was correlated with the phosphorylation and nuclear translocation of nuclear factor-erythroid-2 related factor 2 (Nrf2). However, the cytoprotective and anti-apoptotic effects of anthocyanins were significantly attenuated by the HO-1 inhibitor, demonstrating that anthocyanins promoted Nrf2-induced HO-1 activity to prevent ARPE-19 cells from oxidative stress. Therefore, our findings suggest that anthocyanins, as Nrf2 activators, have potent ROS scavenging activity and may have the potential to protect ocular injury caused by oxidative stress.

• Journal of the Korean Electrochemical Society
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• v.6 no.1
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• pp.59-64
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• 2003

We fabricated mixed ionic-electronic conducting membranes, $CH_4\;Using\;{0.7}Sr_{0.3}Ga_{0.6}Fe_{0.4}O_{3-\delta}$, by solid state reaction method for solid oxide fuel cell. The membranes consisted of single perovskite phase and exhibited high relative density, $>95\%$. We coated $La_{0.6}Sr_{0.4}CoO_{3-\delta}$ layer using screen printing method in order to improve surface reactivity of the $La_{0.7}Sr_{0.3}Ga_{0.6}Fe_{0.4}O_{3-\delta}$. As a result, the oxygen permeation flux of the coated $La_{0.7}Sr_{0.3}Ga_{0.6}Fe_{0.4}O_{3-\delta}$ showed higher value, $0.5ml/min{\cdot}cm^2\;at\;950^{\circ}C$ than the uncoated one. Higher oxygen permeation was observed in the porously coated Lao $La_{0.7}Sr_{0.3}Ga_{0.6}Fe_{0.4}O_{3-\delta}$membranes with larger grain sizes. Syngas, $CO+H_2$, was successfully obtained from methane gas, $CH_4$, using the $La_{0.6}Sr_{0.4}CoO_{3-\delta}$ coated $La_{0.7}Sr_{0.3}Ga_{0.6}Fe_{0.4}O_{3-\delta}$, with over $40\%\;of\;CH_4$ conversion and syngas yield. $La_{0.7}Sr_{0.3}Ga_{0.6}Fe_{0.4}O_{3-\delta}$ membrane was stable even when it was exposed to the reducing environment, methane, for 600 hrs at $950^{\circ}C$.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.36 no.4
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• pp.303-314
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• 2010

In this study, the antibacterial, antioxidative effect and component analysis of Pinus koraiensis leaf extracts were investigated. MIC values of the ethyl acetate fraction from P. koraiensis leaf extracts on P. acnes, S. aureus, P. ovale, and E. coli were 0.06 %, 0.25 %, 0.13 % and 0.50 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction on P. acnes, P. ovale. and S. aureus was more prominent. The aglycone fraction of P. koraiensis leaf extracts ($22.93\;{\mu}g/mL$) showed more higher free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. koraiensis leaf extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The 50 % ethanol extract ($0.70\;{\mu}g/mL$) showed the most prominent ROS scavenging activity. Also the ethyl acetate ($1.04\;{\mu}g/mL$) and the aglycone fraction ($1.43\;{\mu}g/mL$) showed very high antoxidant activity. The protective effects of extract/fractions of P. koraiensis leaf extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The P. koraiensis leaf extracts showed cellular membrane protective effects in a concentration dependent manner ($5{\sim}50\;{\mu}g/mL$). TLC and HPLC chromatogram of the ethyl acetate fraction obtained from hydrolysis of P. koraiensis leaf extracts revealed 2 main bands (PK-4, PK-6) and peaks (peak 1, peak 2), which were identified as kaempferol-3-O-glucoside (PK-6, peak 1) and kaempferol-3-O-arabinoside (PK-4, peak 2) by LC/ESI-MS/MS, respectively. These results indicate that extract/fractions of P. koraiensis can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ROS, and protect cellular membrane against ROS. Extract/fractions of P. koraiensis can be applicable to new cosmeceuticals for antioxidant, antiaging, and antibacterial activity.

• 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.

• 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 Biomedical Engineering Research
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• v.19 no.3
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• pp.279-284
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• 1998

Deep hypothermic circulatory arrest(DHCA), in which systemic temperatures of 2$0^{\circ}C$ or less are used to allow temporary cessation of the circulation, is an useful adjunct in cardiac surgery. Because man in natural circumstances is never exposed to the extreme hypothermic condition, however, one of the controversial aspects is appropriate blood gas management($\alpha$STAT versus PH-STAT) during DHCA. This study aims to compare $\alpha$STAT with PH-STAT management for control of blood gases in experimental cardiopulmonary bypass(CPB) circuits with a membrane oxygenator. Fourteen young pigs were assigned to one of two strategies of gas manipulation. After a median sternotomy, CPB was established. Core cooling was initiated and continued until nasopharyngeal temperature fell below 2$0^{\circ}C$. The flow rate was set at 2,500 ml/min. Once their temperatures were below 2$0^{\circ}C$, the animals were subjected to circulatory arrest for 40mins. During cooling, blood gas was maintained according to either $\alpha$$\alpha$STAT or pH-STAT strategies. After DHCA, the body was rewarmed to normal body temperature. Arterial blood gases were measured before the onset of CPB, before cooling, before DHCA, at the point of 27$^{\circ}C$ during re-warming, on completion of re-warming. Cooling time was significantly shorter in $\alpha$-STAT than PH-STAT strategy, while there was no significant differences in rewarming time between two groups. Carbon dioxide was added between 5.5 and 3.0% in PH-STAT, while no carbon dioxide was added in $\alpha$STAT management. Amounts of oxygen administration were gradually lowered as temperature decreased. In this way, criteria of PH, PaCO, and PaO adjustments were satisfied in both $\alpha$STAT and PH-STAT management groups.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.3
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• pp.225-236
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• 2012

In this study, the cellular protective effects on HaCaT cells and human erythrocytes and antioxidative effects of P. tricuspidata stem extracts were investigated. The ethyl acetate ($50{\mu}g/mL$) and aglycone fraction ($25{\mu}g/mL$) of P. tricuspidata stem extracts doesn't show any characteristics of cytotoxicity. When HaCaT cells were treated with 10 mM $H_2O_2$ and $30{\mu}M$ rose bengal, the ethyl acetate ($6.25{\sim}50{\mu}g/mL$) and aglycone ($6.25{\sim}25{\mu}g/mL$) fraction protected the cells against the oxidative damage in a concentration dependent manner. The P. tricuspidata stem extracts showed more prominent cellular protective effect than (+)-${\alpha}$-tocopherol, known as lipid antioxidant at $10{\mu}g/mL$. The ethylacetate fraction of P. tricuspidata stem extracts ($18.5{\mu}g/mL$) showed more free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC5_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. tricuspidata stem extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate ($1.72{\mu}g/mL$) and the aglycone fraction ($1.53{\mu}g/mL$) showed similar ROS scavenging activity of L-ascorbic acid ($1.50{\mu}g/mL$). These results indicate that extract/fractions of P. tricuspidata stem extracts can function as natural cytoprotective agents and antioxidants in biological systems, particularly skin exposed to UV radiation by protecting cellular membrane against ROS.