• Herbal Formula Science
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• v.26 no.3
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• pp.207-221
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• 2018

Objectives : Present study investigated hepatoprotective effect of Haegan-jeon extract (HE) and tried to elucidate molecular mechanism involved. According to molecular mechanism, present study optimized herbal composition of HE (op-HE) and compared in vitro and in vivo hepatoprotective effects of op-HE to HE. Methods : For in vitro experiments, HepG2 cells were exposed to arachidonic acid (AA, $10{\mu}M$) and iron ($5{\mu}M$) for inducing oxidative stress. Cell viability, GSH contents, $H_2O_2$ production, mitochondrial membrane potential, immunoblot and reporter gene assay were performed to investigate cytoprotective effects and responsible molecular mechanisms. For in vivo experiments, hepatoprotective effect of HE and op-HE were assessed on $CCl_4-induced$ liver injury mice model. Results : HE pretreatment prevented AA+iron-mediated hepatocytes apoptosis. In addition, AA+iron-induced mitochondrial dysfunction, $H_2O_2$ production, glutathione depletion were reduced by HE pretreatment. In addition, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation, antioxidant response element (ARE)-driven reporter gene activity, and antioxidant genes expression were increased by HE. Based on reporter gene and MTT assays, we found that op-HE consisting three medicinal herbs also significantly increased transactivation of Nrf2 and reduced the AA+iron-mediated cytotoxicity. Moreover, in $CCl_4-induced$ liver injury mice model, HE-op had an ability to ameliorate $CCl_4-mediated$ increases in serum alanine transferase and aspartate aminotransferase activity, hepatic degeneration, inflammatory cell infiltration, and collagen deposition. Hepatoprotective effects of op-HE were comparable to those of HE. Conclusions : Present study suggests that op-HE as well as HE exhibit hepatoprotective effect against oxidative stress-mediated liver injury via Nrf2 activation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.10
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• pp.1317-1323
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• 2009

In the present study, we investigated the anti-proliferation activity of Citrus grandis Osbeck peel (CGP) in HL60 (human promyelocytic leukemia) cells. It was found that 80% ethanol extract of CGP could inhibit the cell growth in a dose-dependent manner ($250{\sim}1,000{\mu}g/mL$), which was associated with morphological changes and apoptotic cell death such as depolarized mitochondrial membrane, formation of apoptotic bodies and increased populations of apoptotic sub-G1 phase. The results indicate that CGP extract inhibits the growth of HL60 cancer cells by the induction of apoptosis, which may be mediated by its ability to change the Bcl family proteins and increase the activation of caspase-3 and PARP. Therefore, it is suggested that CGP has the potential to provide a remarkable natural defense against the proliferation of HL60 cells.

• The Journal of Internal Korean Medicine
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• v.23 no.2
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• pp.181-190
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• 2002

The water extract of Omija-tang(OMJT) has been traditionally used for treatment of ischemic heart and brain damage in oriental medicine. However, little is known about the mechanism by which the water extract of OMJT protects cells from such damage. Therefore, this study was conducted to investigate the protective mechanisms of OMJT on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Treatment of $H_2O_2$ markedly induced death of H9c2 cardiomyoblast cells in a dose-dependent manner. The characteristics of $H_2O_2$-induced death of H9c2 showed apparent apoptotic features, such as DNA fragmentation. However, OMJT significantly reduced both $H_2O_2$-induced cell death and chromatin fragmentation. The decrease of Bcl-XL expression by $H_2O_2$ was inhibited by OMJT. In addition, the increase of Bcl-XS and Bax expression were also inhibited by OMJT. In particular, Fas expression, which is generally recognized as cell death inducing signal by Fas/FasL interaction, was markedly increased by $H_2O_2$ in a time-dependent manner, whereas this increase was completely prevented by OMJT. The combined treatment of OMJT and $H_2O_2$ in H9c2 cells also reduced activation of caspase-9 and caspase-3 like protease. Taken together, this study indicates that the protective effects of the water extract of OMJT against oxidative damage may be mediated by the modulation of BcI-XL/S and Bax expression by way of the regulation of mitochondrial membrane potential and caspase cascades.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.633-643
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• 2017

To examine the pro-apoptotic role of the human ortholog (YPEL5) of the Drosophila Yippee protein, the cell viability of Saccharomyces cerevisiae mutant strain with deleted MOH1, the yeast ortholog, was compared with that of the wild-type (WT)-MOH1 strain after exposure to different apoptogenic stimulants, including UV irradiation, methyl methanesulfonate (MMS), camptothecin (CPT), heat shock, and hyperosmotic shock. The $moh1{\Delta}$ mutant exhibited enhanced cell viability compared with the WT-MOH1 strain when treated with lethal UV irradiation, 1.8 mM MMS, $100{\mu}M$ CPT, heat shock at $50^{\circ}C$, or 1.2 M KCl. At the same time, the level of Moh1 protein was commonly up-regulated in the WT-MOH1 strain as was that of Ynk1 protein, which is known as a marker for DNA damage. Although the enhanced UV resistance of the $moh1{\Delta}$ mutant largely disappeared following transformation with the yeast MOH1 gene or one of the human YPEL1-YPEL5 genes, the transformant bearing pYES2-YPEL5 was more sensitive to lethal UV irradiation and its UV sensitivity was similar to that of the WT-MOH1 strain. Under these conditions, the UV irradiation-induced apoptotic events, such as FITC-Annexin V stainability, mitochondrial membrane potential (${\Delta}{\psi}m$) loss, and metacaspase activation, occurred to a much lesser extent in the $moh1{\Delta}$ mutant compared with the WT-MOH1 strain and the mutant strain bearing pYES2-MOH1 or pYES2-YPEL5. These results demonstrate the functional conservation between yeast Moh1 and human YPEL5, and their involvement in mitochondria-dependent apoptosis induced by DNA damage.

• Journal of Life Science
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• v.25 no.11
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• pp.1255-1264
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• 2015

Cnidium officinale, a traditional herb, has diverse beneficial pharmacological activities, such as anti-inflammatory, antioxidant, anticancer, and antiangiogenesis effects. However, the cellular and molecular mechanisms of apoptosis by C. officinale are poorly defined. The present study investigated the proapoptotic effects of water, ethanol, and methanol extract of C. officinale (WECO, EECO, and MECO, respectively) in human leukemia U937 cells. The antiproliferative activity of EECO was higher than that of WECO and MECO. The antiproliferative effect of EECO treatment in U937 cells was associated with the induction of apoptotic cell death, including increased populations of annexin-V positive cells, the formation of apoptotic bodies, DNA fragmentation, and increased numbers of cells with a loss of mitochondrial membrane potential (MMP, Δψm). EECO-induced apoptotic cell death was associated with upregulation of death receptor 4 (DR4) and down-regulation of cellular inhibitor of apoptosis protein-1 (cIAP-1), Bcl-2, and total Bid. The EECO treatment also induced the proteolytic activation of caspases (-3, -8, and -9), and degradation of caspase-3 substrate proteins, such as poly(ADP-ribose) polymerase (PARP), β-catenin, and phospholipase C-γ1 (PLCγ1). In addition, the EECO treatment effectively activated the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway. However, compound C, a specific inhibitor of AMPK, significantly reduced EECO-induced apoptosis. These results indicate that AMPK is a key regulator of apoptosis in response to EECO in human leukemia U937 cells.

• Herbal Formula Science
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• v.25 no.1
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• pp.51-68
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• 2017

Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.

• KSBB Journal
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• v.21 no.4
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• pp.255-259
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• 2006

The biodegradable hyaluronic acid(HA) membranes cross-linked with lactide using the crosslinking agent, 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide(EDC) were prepared as a potential biocompatible material for tissue engineering. HA membranes having different mechanical properties were synthesised by varying degree of the mole ratio of lactide to HA, EDC concentration, and crosslinking temperature. HA membranes were degradable in water solution and the degradation became slower with the increasing mole ratio of lactide to HA. HA membranes were sterilized using ethylene oxide gas and extracted with cell culture medium for 24 h at $37^{\circ}C$ and 200 rpm. Cytotoxicity of the extract was tested using NIH/3T3 mouse embryo fibroblast as a model cell. Growth inhibition was not observed in the extracts of HA membranes with the mole ratios of lactide to HA, 5 or 10, and 10% EDC concentration, however 11% of growth inhibition was observed in the extract with the mole ratio of 13. Growth inhibition was not observed in the extracts of HA membranes prepared with 5% EDC or 10% EDC and the mole ratio of lactide to HA, 10, however 12% of growth inhibition was observed in the extract with 20% EDC. Cytotoxicity was not observed in the extracts of HA membranes prepared at varying crosslinking temperatures, $15^{\circ}C,\;25^{\circ}C,\;and\;28^{\circ}C$ with the mole ratio of lactide to HA, 10 and 10% EDC.

• Journal of Korean Society of Forest Science
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• v.94 no.1 s.158
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• pp.16-20
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• 2005

Tricholoma matsutake ectomycorrhizas are unique in their morphology: not bifurcated broom shaped roots with not easily wettable brilliant and profuse white hyphae. To understand these characteristics the ectomycorrhizas were investigated with electron microscopy. T. matsutake ectomycorrhiza have thin mantle and typical Hartig net development in the epidermis and cortex, but no fungal mantle on the root apex. There were no penetrating hyphae inside of the cells of either epidermis, cortex or stele. Inside of the walls of epidermis and cortex cells are lined with ca. $2{\mu}m$ hemispherical amyloplasts. The brilliant hyphal surface was covered with various fine amorphous granules. The hyphal cell wall was thin membrane less than $0.3{\mu}m$ thick. There is no clamp connection on the hyphae. This thin membraneous cell wall with high elasticity can be related to survival strategy of the species without plasmolysis under frequent soil water stress environment. And the coarse hyphal surface with some water repellency can control sudden inrush of water of the hyphae with an extremely low osmotic potential. It is concluded that no mantle on the tip can induce mycorrhizas not bifurcated and that finely coarse surface of T. matsutake hyphae can make the hyphae brilliantly white but less wetted.

• Journal of Microbiology
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• v.45 no.5
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• pp.460-465
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• 2007

This study was carried out in order to investigate the potential of using plant oils derived from Leptospermum petersonii Bailey and Syzygium aromaticum L. Merr. Et Perry as natural antifungal agents. The antifungal effects of essential oils at concentrations of 0.05, 0.1, 0.15, and 0.2 mg/ml on the dermatophytes Microsporum canis (KCTC 6591), Trichophyton mentagrophytes (KCTC 6077), Trichophyton rubrum (KCCM 60443), Epidermophyton floccosum (KCCM 11667), and Microsporum gypseum were evaluated using the agar diffusion method. The major constituents of the active fraction against the dermatophytes were identified by gas chromatography-mass spectrometry and high-performance liquid chromatography analysis. The antifungal activities of S. aromaticum oil (clove oil) against the dermatophytes tested were highest at a concentration of 0.2mg/ml, with an effectiveness of more than 60%. Hyphal growth was completely inhibited in T. mentagrophytes, T. rubrum, and M. gypseum by treatment with clove oil at a concentration of 0.2 mg/ml. Eugenol was the most effective antifungal constituent of clove oil against the dermatophytes T. mentagrophytes and M. canis. Morphological changes in the hyphae of T. mentagrophytes, such as damage to the cell wall and cell membrane and the expansion of the endoplasmic reticulum, after treatment with 0.11 mg/ml eugenol were observed by transmission electron microscopy (TEM). At a concentration of 0.2 mg/ml, L. petersonii oil (LPO) was more than 90% effective against all of the dermatophytes tested, with the exception of T. rubrum. Geranial was determined to be the most active antifungal constituent of L. petersonii oil. Taken together, the results of this study demonstrate that clove and tea tree oils exhibited significant antifungal activities against the dermatophytes tested in this study.

• Food Science of Animal Resources
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• v.26 no.3
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• pp.402-409
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• 2006

Although microorganisms are, in fact, the most diverse and abundant type of organism on Earth, the ecological functions of microbial populations remains poorly understood. A variety of bacteria including marine Vibrios encounter numerous ecological challenges, such as UV light, predation, competition, and seasonal variations in seawater including pH, salinity, nutrient levels, temperature and so forth. In order to survive and proliferate under variable conditions, they have to develop elaborate means of communication to meet the challenges to which they are exposed. In bacteria, a range of biological functions have recently been found to be regulated by a population density-dependent cell-cell signaling mechanism known as quorum-sensing (QS). In other words, bacterial cells sense population density by monitoring the presence of self-produced extracellular autoinducers (AI). N-acylhomoserine lactone (AHL)-dependent quorum-sensing was first discovered in two luminescent marine bacteria, Vibrio fischeri and Vibrio harveyi. The LuxI/R system of V. fischeriis the paradigm of Gram-negative quorum-sensing systems. At high population density, the accumulated signalstrigger the expression of target genes and thereby initiate a new set of biological activities. Several QS systems have been identified so far. Among them, an AHL-dependent QS system has been found to control biofilm formation in several bacterial species, including Pseudomonas aeruginosa, Aeromonas hydrophila, Burkholderia cepacia, and Serratia liquefaciens. Bacterial biofilm is a structured community of bacterial cells enclosed in a self-produced polymeric matrix that adheres to an inert or living surface. Extracellular signal molecules have been implicated in biofilm formation. Agrobacterium tumefaciens strain NT1(traR, tra::lacZ749) and Chromobacterium violaceum strain CV026 are used as biosensors to detect AHL signals. Quorum sensing in lactic acid bacteria involves peptides that are directly sensed by membrane-located histidine kinases, after which the signal is transmitted to an intracellular regulator. In the nisin autoregulation process in Lactococcus lactis, the NisK protein acts as the sensor for nisin, and NisR protein as the response regulator activatingthe transcription of target genes. For control over growth and survival in bacterial communities, various strategies need to be developed by which receptors of the signal molecules are interfered with or the synthesis and release of the molecules is controlled. However, much is still unknown about the metabolic processes involved in such signal transduction and whether or not various foods and food ingredients may affect communication between spoilage or pathogenic bacteria. In five to ten years, we will be able to discover new signal molecules, some of which may have applications in food preservation to inhibit the growth of pathogens on foods.