• Korean Journal of Microbiology
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• v.42 no.4
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• pp.319-325
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• 2006

[ $\beta$ ]-Glucans (AG) were prepared from Agaricus blazei cultured in the medium fortified with the roots of Pueraria spp. by repeated extraction with hot water, gel filtration chromatography and DEAE ion exchange chromatography. Oligosaccharides (AO) were derived from the hydrolysis of AG by an endo-$\beta$-(1$\rightarrow$6)-glucanase from Bacillus megaterium. The anti-HT-29 human colon cancer activity of AG or AO was investigated using MTT assay, apoptosis assay, cell cycle analysis, and cDNA microairay. AG and AO both inhibited proliferation and growth of HT-29 cells, and stimulated apoptosis of the cells in a dose-dependent manner. In cell cycle analysis, treating HT-29 cells with AG or AO resulted in the increase of cells in the G0 (sub-G1) and G1 phase. Especially, AO was more effective in inducing G0/G1 cell cycle arrest than AG. To screen the genes involved in the increase of apoptosis, the gene expression profile of the HT-29 cells treated with AO was examined by cDNA microarray. While several genes involved in cell cycle progression (CCND2 and CDK2) were down-regulated, many genes involved in apoptosis (TNFSF9, TNFRSF9, FADD, CASP8, BAD, CRADD, CASP9 etc), cell cycle inhibitor (CDKN2A), immune response (IL6, IL18, IL6R etc), and tumor suppressor (CEACAM1, TP53BP2, IRF1, and PHB) were up-regulated. These results suggest that AO could inhibit the proliferation and growth of HT-29 cells by G0/G1 cell cycle arrest and induction of apoptosis.

• Korean Journal of Plant Resources
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• v.31 no.2
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• pp.124-135
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• 2018

The study was performed to explore the molecular changes in the vegetative stage (3-and 5-leaf) of sorghum under waterlogging stress. A total of 74 differentially expressed protein spots were analyzed using LTQ-FT-ICR MS. Among them, 12 proteins were up-regulated and 3 proteins were down-regulated. Mass spectrometry (MS) results showed that about 50% of the proteins involved in various metabolic processes. The level of protein expression of malate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase related to carbohydrate metabolic process increased in both 3 and 5-leaf stage under waterlogging stress. These proteins are known to function as antistress agents against waterlogging stress. The expression of oxygen-evolving enhancer protein 1 protein related to photosynthesis was slightly increased in the treated group than in the control group, however the expression level was increased in the 5-leaf stage compared to the 3-leaf stage. Probable phospholipid hydroperoxide glutathione peroxidase protein and superoxide dismutase protein related to response to oxidative stress showed the highest expression level in 5-leaf stage treatment. This suggests that the production of reactive oxygen species by the waterlogging stress was the most abundant in the 5-leaf treatment group, and the expression of the antioxidant defense protein was increased.

• Restorative Dentistry and Endodontics
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• v.31 no.6
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• pp.437-444
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• 2006

This study was aimed to elucidate the biological function of OD314 (Apin protein), which is related to ameloblast differentiation and amelogenesis. Apin protein, calcifying epithelial odontogenic (pindborg) tumors (CEOTs)-associated amyloid, were isolated from CEOTs, and has similar nucleotide sequences to OD314. We examined expression of the OD314 mRNA using in-situ hybridization during tooth development in mice. Expression of OD314 and several enamel matrix proteins were examined in the cultured ameloblast cell line up to 28 days by reverse transcription-polymerase chain reaction (RT-PCR) amplification. After inactivation and over-expression of the OD314 gene in ameloblast cell lines using U6 vectordriven RNA interference and CMV-OD314 construct, RT-PCR were performed to evaluate the effect of the OD314 during amelogenesis. The results were as follows: 1. In in-situ hybridization, OD314 mRNAs were more strongly expressed in ameloblast than odontoblast. 2. When ameloblast cells were cultured in the diffcrentiation and mineralization medium for 28 days, the tuftelin mRNA expression was maintained from the beginning to day 14, and then gradually decreased to day 28. The expressions of amelogenin and enamelin were gradually decreased according to the ameloblast differentiation. 3. Inactivation of OD314 by U6-OD314 siRNA construct down-regulated the expression of OD314, MMP-20, and tuftelin, whereas over-expression of OD314 by CMV-OD314 construct up-regulated the expression of OD314 and MMP-20 without change in tuftelin. These results suggest that OD314 is considered as an ameloblast-enriched gene and may play the important roles in ameloblast differentiation and mineralization.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.257-267
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• 2017

Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.5
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• pp.655-662
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• 2013

The antioxidant and anticancer effects of the edible mushrooms Lentinus edodes (LE, Pyogo mushroom) and Agaricus blazei (AB, Agaricus mushroom), and the medicinal mushrooms Cordyceps militaris (CM, Dong chunghacho), Ganoderma lucidum (GL, Youngji mushroom), Inonotus obliquus (IO, Chaga mushroom), and Phellinus linteus (PL, Sangwhang mushroom) were studied in vitro. The bioactive components were extracted by methanol. The antioxidant effects were evaluated using the DPPH and hydroxyl radical scavenging assays. The antioxidant activities of medicinal mushrooms (35~90%) were higher than edible mushrooms (4~23%). The in vitro anticancer effects of the mushrooms were evaluated using the MTT assay in AGS gastric adenocarcinoma cells, HCT-116 colon carcinoma cells, and HepG2 hepatoma cells. The medicinal mushrooms CM, GL, IO, and PL showed 28~91% inhibition, while the edible mushrooms LE and AB exhibited 5~40% inhibition. The medicinal mushrooms, compared to edible mushrooms, effectively down-regulated the gene expression of the anti-apoptosis related gene Bcl-2 and inflammation-related genes iNOS and COX-2, and up-regulated the pro-apoptosis gene Bax (p<0.05). Total polyphenol and flavonoids contents of the medicinal mushrooms were 9.1~35.7 mg/g, while the edible mushrooms showed 0~13.3 mg/g. This study showed that antioxidant activities and anticancer activities in vitro increased in the order LE, AB, GL, CM, IO and PL. LE and AB showed the lowest effects among the samples, GL and CM had medium effects, and IO and PL exhibited the highest effects in the antioxidant and anticancer effect for three different human cancer cells. Taken together, PL resulted in the highest and LE the lowest effects in this study.

• Journal of Life Science
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• v.31 no.2
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• pp.126-136
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• 2021

Oxidative stress causes injury to and degeneration of retinal pigment epithelial (RPE) cells. It is involved in several retinal disorders and leads to vision loss. In the present study, we investigated the effect of 14 kinds of natural compounds and two kinds of synthetic compounds on oxidative stress-induced cellular damage in human PRE cell lines (ARPE-19). From among them, we selected five kinds of compounds, including auranofin, FK-509, hemistepsin A, honokiol, and spermidine, which have inhibitory effects against hydrogen peroxide (H2O2)-mediated cytotoxicity. In addition, we found that four kinds of compounds (excluding auranofin) have protective effects on H2O2-induced mitochondrial dysfunction. Furthermore, the expression of phosphorylation of histone H2AX, a sensitive marker of DNA damage, was markedly up-regulated by H2O2, whereas it was notably down-regulated by FK-506, honokiol, and spermidine treatment. Meanwhile, five kinds of candidate compounds had no effect on H2O2-induced intracellular reactive oxygen species (ROS) levels, suggesting that the five candidate compounds have protective effects on oxidative stress-induced cellular damage through the ROS-independent pathway. Taken together, according to the results of H2O2-mediated cellular damage―such as cytotoxicity, apoptosis, mitochondrial dysfunction, and DNA damage―spermidine and FK-506 are the natural and synthetic compounds with the most protective effects against oxidative stress in RPE. Although further studies on the identification of the mechanism responsible are required, the results of the present study suggest the possibility of using spermidine and FK-506 to suppress the risk of retinal disorders.

• Archives of Pharmacal Research
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• v.25 no.6
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• pp.910-916
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• 2002

Endotoxin tolerance reduces the capacity of monocytes to produce proinflammatory cytokines, results in cellular immune paralysis, and down-regulates the production of helper T (Th)1 type cytokines with a shift toward a Th2 cytokine response. Prostaglandin (PG)E$_2$ in the immune system also results in macrophage inactivation and the suppression of Th1 activation and the enhancement of Th2 activation. However, the inhibitory effects of PGE$_2$ on the altered polarization of the Th cell and macrophage interleukin (IL)-6 production characterized in part by cellular immune paralysis in a state of endotoxin tolerance is unclear. This study was undertaken, using indomethacin, to investigate the role of endogenous PGE$_2$ on the Th cytokines and macrophage IL-6 production in a state of endotoxin tolerance compared to those with endotoxemia mice, wherein, in this latter case, the increased production of proinflammatory cytokines and PGE$_2$ is exhibited. Endotoxemia was induced by injection of lipopolysaccharide (LPS; 10 mg/kg in saline) i.p. once in BALB/c mice, and endotoxin tolerance was induced by pretreatment with LPS (1 mg/kg in saline) injected i.p. daily for two consecutive days and then with LPS 10 mg/kg on day 4. Splenocytes or macrophages were obtained from endotoxemia and endotoxin tolerance models pretreated with indomethacin, and then cytokine production was induced by Con A-stimulated splenocytes for the Th cytokine assays and LPS-stimulated macrophages for the IL-6 assay. Our results showed that endotoxemia led to significantly reduced IL-2 and IL-4 production, to significantly increased IL-6 production, whereas interferon $(IFN)-{\gamma}$ production was not affected. Indomethacin in the case of endotoxemia markedly attenuated $IFN-{\gamma}$ and IL-6 production and didnt reverse IL-2 and IL-4 production. Endotoxin tolerance resulted in the significantly reduced production of IL-2 and $IFN-{\gamma}$ and the significantly increased production of IL-4 and IL-6. Indomethacin in endotoxin tolerance greatly augmented IL-2 production, significantly decreased IL-4 production, and slightly attenuated IL-6 production. These findings indicate that endogenous PGE$_2$ may mediate the suppressed Th1 type immune response, with a shift toward a Th2 cytokine response in a state of endotoxin tolerance, whereas endotoxemia may be regulated differentially. Also, endogenous PGE$_2$ may mediate macrophage IL-6 production in the case of endotoxemia to a greater extent than in the case of endotoxin tolerance.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.532-539
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• 2015

Inhibition of osteoclast differentiation is the most important target for prevention of inflammatory bone resorption and bone diseases. Here, we investigated the effect of spinach ethanol extract on osteoclast differentiation in RAW264.7 cells. Spinach was extracted with ethanol at a concentration ranging from 0 to 100% (0, 25, 50, 75, and 100% ethanol). Inhibitory effects of receptor activator of NF-${\kappa}B$ ligan (RANKL)-induced osteoclast differentiation were evaluated using tartrate-resistant acid phosphatase (TRAP) stain assay. The most effective eanol concentration for osteoclast differentiation was 100%. Spinach extract (100% ethanol) suppressed RANKL-induced osteoclast differentiation and TRAP activity. Spinach extract (100% ethanol) also suppressed expression of osteoclast differentiation-related marker genes (NFATc1, c-FOS, cathepsin K, and TRAP) and down-regulated RANKL-induced NF-${\kappa}B$ and ERK phosphorylation during osteoclast differentiation. Taken together, our results suggest that spinach extract is effective against reducing osteoclast differentiation through the NF-${\kappa}B$-mediated pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.12
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• pp.1708-1714
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• 2011

This study investigated the possible anti-obesity effects of Hypsizigus marmoreus on high fat-fed mice. Thirty male C57BL/6 mice were randomly divided into 3 groups: a normal diet group (N), a high-fat diet group (HF), and a high-fat diet with 5% Hypsizigus marmoreus group (HF-H). After 8 weeks, the body weights in the HF group significantly increased, while those of the HF-H group decreased. Also, liver and adipose tissue weights in the HF-H group significantly decreased. Total serum cholesterol, leptin, and insulin levels were significantly higher in the HF group than those of the N group, but lower than those of the HF-H group. Accumulation of hepatic lipids was apparent in the HF group, as indicated by HE staining and hepatic lipid analysis, while these effects were improved by supplements with Hypsizigus marmoreus in the HF-H group. Also, a reduction in adipocyte size of the epididymal adipose tissue was observed in the HF-H group. $PPAR{\gamma}$, SREBP-1c, and SCD-1 protein expressions were down-regulated in the epididymal adipose tissue of the HF-F group compared to the HF group. Taken together, these results suggest that Hypsizigus marmoreus may an effective anti-obesity treatment.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.20 no.3
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• pp.168-174
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• 2010

South Korea's industrial injuries are decreasing overall in the last 32 years. Nevertheless, the fatal occupational injury rate is still higher than in developed countries. This study was conducted to help prevention strategies of occupational injuries for the Republic of Korea. Fatal occupational injury rates were obtained from "Industrial Accident Analysis"of the Korean Ministry of Labor. Poisson regression was used to assess time trends. Socioeconomic indicators were obtained from the Korea Labor Institute and the Statistics Korea. Fatal occupational injury rates were adjusted by year, and Pearson correlation analysis was used to assess the relationship between the socio-economic indicators and occupational injuries. In 1975, fatal occupational injury rate was 54.8 per 100,000 workers. With somewhat up and down, it was decreased to 21.0 in 2006. An annual rate of change for the years 1975-2006 was - 1.83%, and for the years 2002-2006 was -5.02%. As economic growth rate, paricipation rate for the age less than 25 and hours of work per week or year increased, fatal occupational injury rate also increased. Conversely, as GDP per capita, paricipation rate or employment rate for female, paricipation rate for the age 25 or more, hourly compensation costs for production workers and services output as percent of GDP increased, fatal occupational injury rate decreased. By the development of safety techniques and the adoption of more legislative constraints, developed economy reduce occupational injuries. Conversely, economic growth may raise occupational injuries. Therefore, prevention strategies are needed to manage both of them. We need to make an effort to prevent occupational injuries due to not only sexual differences, but also job differences between male and female. Preventive strategies are needed to consider the characteristics of younger workers. Addition to wage, other appropriate variables for work condition should be considered together. Extending work hours is need to be regulated with systemic methods.