• Title/Summary/Keyword: Hep-G2 cell

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Antioxidant Activity of Novel Casein-Derived Peptides with Microbial Proteases as Characterized via Keap1-Nrf2 Pathway in HepG2 Cells

  • Zhao, Xiao;Cui, Ya-Juan;Bai, Sha-Sha;Yang, Zhi-Jie;Cai, Miao;Megrous, Sarah;Aziz, Tariq;Sarwar, Abid;Li, Dong;Yang, Zhen-Nai
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1163-1174
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    • 2021
  • Casein-derived antioxidant peptides by using microbial proteases have gained increasing attention. Combination of two microbial proteases, Protin SD-NY10 and Protease A "Amano" 2SD, was employed to hydrolyze casein to obtain potential antioxidant peptides that were identified by LC-MS/MS, chemically synthesized and characterized in a oxidatively damaged HepG2 cell model. Four peptides, YQLD, FSDIPNPIGSEN, FSDIPNPIGSE, YFYP were found to possess high 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability. Evaluation with HepG2 cells showed that the 4 peptides at low concentrations (< 1.0 mg/ml) protected the cells against oxidative damage. The 4 peptides exhibited different levels of antioxidant activity by stimulating mRNA and protein expression of the antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), as well as nuclear factor erythroid-2-related factor 2 (Nrf2), but decreasing the mRNA expression of Kelch-like ECH-associated protein 1 (Keap1). Furthermore, these peptides decreased production of reactive oxygen species (ROS) and malondialdehyde (MDA), but increased glutathione (GSH) production in HepG2 cells. Therefore, the 4 casein-derived peptides obtained by using microbial proteases exhibited different antioxidant activity by activating the Keap1-Nrf2 signaling pathway, and they could serve as potential antioxidant agents in functional foods or pharmaceutic preparation.

N-Terminal Amino Acid Sequences of Receptor-Like Proteins that Bind to preS1 of HBV in HepG2 Cells

  • Lee, Dong-Gun;Liu, Ming-Zhu;Kim, Kil-Lyong;Hahm, Kyung-Soo
    • BMB Reports
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    • v.29 no.2
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    • pp.180-182
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    • 1996
  • One of the essential functions of virus surface proteins is the recognition of specific receptors on target cell membranes, and cellular receptors play an important role in viral pathogenesis. But the earliest steps of hepatitis B virus (HBV) infection, such as hepatocyte receptor interaction with the virus, are poorly understood. Previous work has suggested an important role of the preS1 region of HBV envelope protein in mediating viral binding to hepatocytes. Although hepatitis B virus (HBV) infection appears to be initiated by specific binding of virions to cell membrane structures via one or potentially several viral surface proteins, data showing the identification or isolation of the HBV receptor (s) are not yet available. The receptor-like proteins on the plasma membrane surface of HepG2 cells that bind to PreS1 were separated and identified using affinity chromatography, and the amino-terminal amino acid sequences of the receptor-like proteins were determined.

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Cytolytic Effects of an Adenoviral Vector Containing L-Plastin Promoter Regulated E1A in Hepatocellular Carcinoma Cells

  • Chung, In-Jae
    • Biomolecules & Therapeutics
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    • v.14 no.3
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    • pp.148-151
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    • 2006
  • We have previously reported that 2.4 kb of L-plastin promoter (LP) could regulate the expression of adenoviral vector (AV) exogenous genes in a tumor cell specific manner. In the present study, we tested if the replication competent AdLPE1A vector results in a direct cytotoxic effect in hepatocelluar carcinoma (HCC) cells. In vitro cytotoxicity tests were carried out with replication-competent (AdLPE1A) and -incompetent (AdLPCD) LP-driven vectors. AdLPE1A is an AV in which LP was inserted 5' to the E1A and E1B genes. The AdLPCD vector contains LP and the E. coli cytosine deaminase (CD) gene in transcription unit. Exposure of cells to AdLPE1A generated a significant cytotoxic effect as compared to the control. Almost 90% of the cell had manifested the characteristic cytopatic effect on day 9 after infection of cells with 10 MOI of AdLPE1A. On the other hand, almost 35% of the cells were left when the cells had been treated with 100 MOI of AdLPCD together with 5-FC on day 9 when compared with the cells which had never been exposed neither 5-FC nor AdLPCD. These results showed that the replication competent AdLPE1A vector could kill the HepG2 cells directly by the oncolytic effect of the virus. The replication competent AV vector carrying viral E1A generated greater cytotoxic effect than the replication incompetent AV, which contains the CD prodrug activation transcription unit without E1A, in HepG2 cells.

Isolation and quantitative analysis of metabolites from Scrophularia buergeriana and their hepatoprotective effects against HepG2 Cells (현삼 (Scrophularia buergeriana)에서 분리한 화합물의 함량분석 및 간세포 보호 효과)

  • Na, Hyeon Seon;Oh, Seon Min;Shin, Woo Cheol;Bo, Jeon Hwang;Kim, Hyoung-Geun;Yoon, Dahye;Yang, Seung Hwan;Lee, Young-Seob;Kim, Geum-Soog;Baek, Nam-In;Lee, Moon-Soon;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.399-406
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    • 2019
  • The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H2O fractions. The repeated silica gel or octadecyl SiO2column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

Effect of the Geijibokryunghwan on human hepatocarcinoma cells

  • Lee Soo Kyung;Kim Han Geu;Ahan Jong Chan;Chung Tae Wook;Moon Jin Young;Park Sun Dong;Kim June Ki;Choi Dall Yeong;Kim Cherl Ho;Park Won Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.568-573
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    • 2003
  • We invesgated the GBH water extracts can be used as a potential cancer chemopreventive agent in humans, especially in hepatological cancer cell lines. The GBH was found to act as an potent inhibitor of COX-I only, but not as COX-2 inhibitor. Furthermore, the extract mediated anti-inflammatory effects and inhibited COX-associated hydroperoxidase functions(antipromotion activity). Inhibitory effect of the GBH water extracts on the growth of cancer cell lines such as HepG2 cell and Hep3B cell was shown.

Inhibitory effects of Sargassum horneri extract against endoplasmic reticulum stress in HepG2 cells (괭생이 모자반 추출물의 소포체 스트레스 억제 효능)

  • Park, Sora;Thomas, Shalom Sara;Cha, Youn-Soo;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.53 no.6
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    • pp.583-595
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    • 2020
  • Purpose: This study examined the effects of Sargassum horneri extracts on palmitic acid (PA)-induced endoplasmic reticulum (ER) stress in HepG2 cells. Methods: HepG2 cells were treated with varying concentrations of S. horneri extract or PA, and the cell viability was measured by water soluble tetrazolium salts analysis. The effective induction of ER stress and the effects of S. horneri were investigated through an examination of the ER stress-related genes, such as activating transcription factor 4 (ATF4), X-box binding protein (XBP1s), C/EBP homologous protein (CHOP), and 78-kDa glucose-regulated protein (GRP78) by quantitative reverse transcription polymerase chain reaction. The expression and activation levels of unfolded protein response (UPR) associated proteins, such as inositol-requiring enzyme-1α (IRE1α), eukaryotic translation initiation factor 2 alpha submit (eIF2α), and CHOP were examined by western blot analysis. Results: The treatment with PA increased the expression of UPR associated genes significantly and induced ER stress in a 12-hour treatment. Subsequent treatment with S. horneri reduced mRNA expression of ATF4, GRP78, and XBP1s. In addition, the protein levels of phosphate (p)-IRE1α, p-elF2α, and CHOP were also reduced by a treatment with S. horneri. An analysis of sirtuin (SIRT) mRNA expression in the S. horneri and PA-treated HepG2 cells showed that S. horneri increased the levels of SIRT2, SIRT6, and SIRT7, which indicates a possible role in reducing the expression of ER stress-related genes. Conclusion: These data indicate that S. horneri can exert an inhibitory effect on ER stress caused by PA and highlight its potential as an agent for managing various ER stress-related diseases.

Alteration in miRNA Expression Profiling with Response to Nonylphenol in Human Cell Lines

  • Paul, Saswati;Kim, Seung-Jun;Park, Hye-Won;Lee, Seung-Yong;An, Yu-Ri;Oh, Moon-Ju;Jung, Jin-Wook;Hwang, Seung-Yong
    • Molecular & Cellular Toxicology
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    • v.5 no.1
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    • pp.67-74
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    • 2009
  • Exposures to environmental chemicals that mimic endogenous hormones are proposed for a number of adverse health effects, including infertility, abnormal prenatal and childhood development and above all cancers. In addition, recently miRNA (micro RNA) has been recognized to play an important role in various diseases and in cellular and molecular responses to toxicants. In this study, endocrine disrupting environmental toxicant, nonylphenol (NP) was treated to MCF-7 (Human breast cancer cell) and HepG2 (Human hepatocellular liver carcinoma) cell line at 3 hrs and 48 hrs time point and miRNA analysis using $mirVana^{TM}$ miRNA bioarray was performed and compared with total mRNA microarray data for the same cell line and treatment. Robust data quality was achieved through the use of dye-swap. Analysis of microarray data identifies a total of 20 and 11 miRNA expressions at 3 hrs and 48 hrs exposure to NP in MCF-7 cell line and a total of 14 and 47 miRNA expression at 3 hrs and 48 hrs exposure respectively to NP in HepG2 cell line. Expression profiling of the selected miRNA (let-7c, miR-16, miR-195, miR-200b, miR200c, miR-205, and miR-589) reveals changes in the expression of target genes related to metabolism, immune response, apoptosis, and cell differentiation. The present study can be informative and helpful to understand the role of miRNA in molecular mechanism of chemical toxicity and their influence on hormone dependent disease. Also this study may prove to be a valuable tool for screening potential estrogen mimicking pollutants in the environment.

Comparison of Antioxidant Activities of Water Extract from Dandelion (Taraxacum officinale) Aerial Parts, Roots, and Their Mixtures (서양민들레(Taraxacum officinale) 지상부, 지하부 및 혼합 추출물의 항산화 활성 비교)

  • Jung, Hyun Jung;Sung, Hea Mi;Kim, Kyung Mi;Shin, Yu-Rim;Wee, Ji-Hyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1157-1164
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    • 2015
  • The present investigation evaluated the antioxidant activities of water extracts from dandelion (Taraxacum officinale) aerial parts, roots, and mixed extracts. Mixed extract of T. officinale was a mixture of aerial parts and roots at 9:1 and 8:2 weight ratios. Extracts from aerial parts (DAE), roots (DRE), and mixture of aerial parts and roots (DME) were measured for cell viability and catalase activity in HepG2 cells, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, and lipid peroxidation inhibitory activity. Cell viabilities of HepG2 cells treated with DAE, DRE, DME 8:2, and DME 9:1 against $H_2O_2$-induced oxidative damage were 63.4%, 54.6%, 76.7% and 83.4% at a concentration of $400{\mu}g/mL$, respectively. Catalase activity was highest in DME 9:1 (12.2 mU/min/mg protein) compared with DAE (9.0 mU/min/mg protein) and DRE (9.7 mU/min/mg protein). DPPH radical scavenging activity of DME showed a significantly lower $EC_{50}$ value than DAE ($EC_{50}$ value of DME $9:1=163.3{\mu}g/mL$, DME $8:2=172.4{\mu}g/mL$, and $DAE=173.7{\mu}g/mL$). Lipid peroxidation inhibitory activity of DME showed a significantly lower $EC_{50}$ value than DAE [$EC_{50}$ values of DME $(9:1)=454.4{\mu}g/mL$, DME $(8:2)=426.6{\mu}g/mL$, and $DAE=654.7{\mu}g/mL$]. The results indicate that a small amount of T. officinale roots increased antioxidant activity of aerial parts. Especially, a 9:1 mixture was more valuable than 8:2 mixture for industry.