• Journal of Environmental Health Sciences
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• v.30 no.3
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• pp.230-238
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• 2004

The adverse health effects of a number of environment pollutions are related to the formation of free radicals. Induction of antioxidant defensive system in the response to an oxidative attack is an essential element of the cell to survive. CYP2E1 is easily induced by organic solvents and induces continuous formation of reactive oxygen species (ROS). ${\gamma}$-Glutamyltranspeptidase (${\gamma}$GT) plays an important role in glutathione metabolism and xenobiotic detoxification. To evaluate the characteristic of oxidative stress which induces GGT expression and to understand human antioxidant defensive response against oxidative stress induced by CYP2E1, we studied regulation of ${\gamma}$GT enzyme expression in response to various oxidative stresses in human HepG2 cells. The ${\gamma}$GT activity was not modified after exposure of acute oxidative stress inducing agents (ferric nitrilotriacetate, cumene hydroperoxide, ADP-Fe, O-tetradecanoylphorbol-13-acetate, tumor necrosis factor-alpha). To induce continuous exposure of cells to ROS, HepG2 cells were transfected by human CYP2E1 gene transiently. The CYP2E1 activity was verified with chlorzoxazone hydroxylation. Transfection of CYP2E1 showed continuous 60% increase in intracellular ROS and 240 % increase in microsomal ROS. CYP2E1 overexpressing cells showed increased ${\gamma}$GT activity (2.5-fold). The observed enhancement of ${\gamma}$GT activity correlated with a significant increase of ${\gamma}$GT mRNA (2.1-fold). Treatment with antioxidant strongly prevented the increase in ${\gamma}$GT activity. The CYP2E1 overexpression did not modify toxicity index and increased glutathione levels. These results show that continuous exposure of cells to ROS produced by CYP2E1 up-regulates ${\gamma}$GT; This may be one of the adaptive antioxidant responses of cells to oxidative insult. Present study also suggests that the induction of ${\gamma}$GT could be used as a marker of oxidative stress induced by exposure to organic solvents.

• Journal of Acupuncture Research
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• v.23 no.3
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• pp.177-190
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• 2006

Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.

• Journal of Ginseng Research
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• v.37 no.4
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• pp.442-450
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• 2013

The aim of this study was to determine and compare the preventive effect of Korean White Ginseng and Red Ginseng on oxidative stress in $H_2O_2$-treated HepG2 cells. The roots of ginseng were extracted with 70% methanol and partitioned with butanol to obtain saponin fractions, which have been known as bioactive constituents of ginseng. 2',7'-Dichlorofluorescein diacetate (DCF-DA) assay and malondialdehyde (MDA) content were measured for evaluating intracellular reactive oxygen species (ROS) generation. Also, mRNA expressions and activities of antioxidant enzymes were analyzed to determine the antioxidant activity of saponin or non-saponin fractions of ginsengs. According to DCF-DA assay, $H_2O_2$-induced MDA release and ROS generation were significantly reduced by treatment with saponin fractions of white and red ginseng roots. Also, saponin fractions increased effectively intracellular antioxidant enzyme activities including catalase, glutathione peroxidase and superoxide dismutase in $H_2O_2$-treated HepG2 hepatoma cells. In general, red ginseng was more effective than white ginseng for reducing oxidative stress. These results indicate that administration of red ginseng may certainly contribute relatively stronger than white ginseng to prevent from damaging liver function by oxidative stress.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.9
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• pp.1121-1126
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• 2006

In the present study, the potential hepatoprotective effects of poly herbal formulation, Hepa-1000, against oxidative damages induced by t-BHP were evaluated in HepG2 cells in order to relate in vitro antioxidant activity with cytoprotective effects. The t-BHP induced considerable cell damage in HepG2 cells was shown by significant glutamic oxaloacetic transaminase (GOT) and lactate dehydrogenase (LDH) leakage, and increased lipid peroxidation. Hepa-1000-treated cells showed an increased resistance to oxidative challenge, as revealed by higher survival capacity than the one of control cells against t-BHP induced oxidative stress and hepatotoxicity. In addition, the Hepa-1000 had hepatoprotective effects lowering the activity of GOT and LDH, simultaneously. That is, it could inhibit the cell membrane damages resulting in the increased activities of GOT and LDH in the cell culture media. Furthermore, the Hepa-1000 could reduce t-BHP enhanced lipid peroxidation, which was evaluated by measuring the production of malonedialdehyde. Based on the data described above, it could be suggested that the Hepa-1000 has significant hepatoprotective effects and plays a protective role against lipid peroxidation by free radicals.

• The Journal of Korean Medicine
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• v.21 no.1
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• pp.53-61
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• 2000

Objectives: This study was camed out to examine the effect of five fractions of aqueous extract from Artemisia capillaris THUNB(ACT), on TGF, 1-induced apoptosis, cell viability, cell cycle progression and mRNA expression of apoptosis-related genes in human hepatocyte cell line HepG2. Methods: This study employed Tryphan blue exclusion assay, DNA fragmentation assay, Cpp32 protease activity assay and Quantitative RT-PCR analysis. Results: In the Tryphan blue exclusion assay, the butanol fraction of ACT with $TGF{\beta}$, l showed magnificent (Nice word, ut is it appropriate in a medical abstract\ulcorner) viability and the H2O fraction of ACT with $TGF{\beta}$, l also showed higher viability than only $TGF{\beta}$, l-treated group. DNA fragmentation assay showed that the butanol fraction and the H2O fraction carried inhibitory effects on apoptosis induction, with the butanol fraction displaying greater effects. The Cpp32 protease activity assay showed that the butanol fraction decreased Cpp32 protease activity. The H2O fraction of ACT had no significant effect on the Cpp32 protease activity. Quantitative RT-PCR showed that the butanol fraction suppressed Bax, p 15/INK4B, p21/Waf1, PAI-1 and increased Bcl-2 gene. Conclusions: The data shows that butanol fraction of ACT increases the hepatocyte viability and has the hepatocellular protective effect by the suppression of $TGF{\beta}$, l induced-apoptosis through gene regulation.

• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.362-367
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• 2003

Coriolus versicolor was grown in a defined synthetic liquid medium and citrus extracts, and the culture extracts were examined for antioxidant activity, nitrite scavenging activity, and in vitro anticancer activity against HeLa, PC-3, HepG2, and A-549 cells. Whereas the culture extracts obtained from the synthetic medium and the un-inoculated citrus extract showed 60 and 22％ of the 1,1-diphenyl-2-picrylhydrazyl radical scavenger activity, the culture extracts obtained from the citrus extracts medium exhibited antioxidant activity up to 89％. The nitrite scavenging activity of the culture extracts obtained from the citrus extracts medium and the synthetic liquid medium, and the un-inoculated citrus extract at pH 1.2 were up to 67, 55, and 34％, respectively. The culture extract obtained from the synthetic liquid medium inhibited the growth of HeLa, PC-3, HepG2, and A-549 up to 66, 23, 18, 10％ at 48 h of incubation, respectively; however, the culture extract obtained from the citrus extracts medium inhibited the growth of HeLa, PC-3, HepG2, and A-549 up to 75, 82, 55, and 82％, respectively. As a negative control, the un-inoculated citrus extract was examined in the same way and inhibited the growth of HeLa, PC-3, and HepG2 cells 20, 6, and 15％ at 48 h incubation, respectively; the inhibition of A-549 cell growth was negligible. These results clearly showed that the fermentation of C. versicolor in the citrus extracts rather than in the defined synthetic medium significantly enhanced the anticancer activity, antioxidant activity, and nitrite scavenging activity.

• Molecular & Cellular Toxicology
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• v.1 no.4
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• pp.224-229
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• 2005

Cirsii Japonici Herba (CJH) extract has been used for hundreds of years in Asian countries as a treatment for pollutant, radiation, and alcohol-induced liver damage. The reducing effect of CJH on hydrogen peroxide-induced reactive oxygen species (ROS) production, the main cause of cell damage or death, was evaluated using the HepG2 cell line. Cell survival was determined using MTS assay. The viability of cells treated with CJH was not significantly different from oxidative-stressed HepG2 cells. A dose-dependent inhibitory effect by CJH on ROS production was shown in oxidative-stressed cells using the $H_{2}DCFDA$ assay. To identify candidate genes responsible for the anti-oxidative effects of CJH on HepG2 cells, an oligonucleotide microarray analysis was performed. The expressions of five genes were decreased, whereas nineteen genes were up-regulated in CJH plus hydrogen peroxide treated cells, compared to only hydrogen peroxide treated cells. Among them, the expression of 5 genes was decreased in hydrogen peroxide treatment when compared to control. These genes are known to regulate cell survival and progression. On the other hand, it was shown that its main compounds were not a sylimarin or its analogs. The list of differentially expressed genes may provide further insight on the action and mechanism behind the anti-oxidative effects of Cirsii Japonici Herba.

• Food Science and Preservation
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• v.24 no.5
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• pp.673-679
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• 2017

In this study, the antimicrobial, antioxidant activities and ${\alpha}$-glucosidase inhibitory activities of Amphidinium carterae ethanol extract (AE) was evaluated for using as a functional food ingredient. Chlorella ethanol extract (CE) was used to the comparison as a control. Anticancer activities of the AE and CE were analyzed by HepG2 and HT-29 human cancer cell. The AE showed antimicrobial activities for all tested bacterial strains. Whereas, CE showed antimicrobial activities for several tested bacterial strains only. The CE showed higher total phenolics contents, DPPH and ABTS radical-scavenging activities (47.36 mg/g, 22.42% and 28.58%, respectively) than those of AE (8.88 mg/g, 20.16% and 17.69%, respectively). AE showed anti-diabetic effect on ${\alpha}$-glucosidase inhibitory activity with dose-dependantly manner. The cell viability of AE ($125{\mu}g/mL$) on HepG2 and HT-29 human cancer cells were 38.12% and 11.27%, respectively. It was demonstrated that ethanol was efficient solvent for extracting functional components from A. carterae. These results indicated that AE can be described as a good candidate for using as a functional food ingredient.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.2
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• pp.113-119
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• 2007

In this study, we investigated antimicrobial and cytotoxicity effects of Gelidium amansii L., which using methanol, dichloromethane and ethanol were extracted and fractionated into four different types : methanol (GAMM), hexane (GAMH), butanol (GAME) and aqueous (GAMA). The antimicrobial activity was increased in proportion to its concentration by the paper disc method. Among the solvent fractions, The methanol partition layer (GAMM) showed the strongest antimicrobial activities and cytotoxic effects on all cancer cell lines. We also observed quinone reductase (QR) induced effects in all fraction layers of GA on HepG2 cells. The QR induced effects of GAMM on HepG2 cells at $40{\mu}g/mL$ concentration indicated 2.5 with a control value of 1.0.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.10
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• pp.1238-1243
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• 2008

The root of Adenophora triphylla is widely used as traditional herbal medicine in Korea. We studied its effects on sodium nitroprusside (SNP) cytotoxicity and antioxidant genes expression in HepG2 cells. To study whether Adenophora triphylla ethylacetate extract (ATea) inhibited NO-induced cell death, HepG2 cells were preincubated for 24 hr with 50 and 100 $\mu$g/mL ATea followed by 24-hr exposure to 0.5 mM SNP (exogenous NO donor). No-induced cytotoxicity was inhibited by pretreatment of ATea, as assessed by mitochondrial dehydrogenase activity (MTT assay). We further investigated the effects of ATea on mRNA levels of various enzymes of the antioxidant system such as Cu, Zn superoxide dismutase (SOD 1), Mn SOD (SOD 2), glutathione peroxidase (GPx), catalase and several enzymes of the glutathione metabolism [glutathione reductase (GR), $\gamma$-glutamyl-cystein synthetase (GCS), glutathione-S-transferase (GST), $\gamma$-glutamyltranspeptidase ($\gamma$-GT), glucose-6-phosphate dehydrogenase (G6PD)] by RT-PCR. CAT, GCS, GR and G6PD mRNA levels were increased after treatment with ATea. The SOD 1, SOD 2, GPx, GST and $\gamma$-GT mRNA levels were not affected in ATea-treated HepG2 cells. We concluded that ATea have an indirect antioxidant effects, perhaps via induction of CAT, GCS, GR and G6PD.