• Title/Summary/Keyword: HepG2-ARE-C8

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Antioxidant and Antigenotoxic Effects of Shiitake Mushrooms Affected by Different Drying Methods (건조 방법에 따른 표고버섯의 항산화능과 항유전독성 효과)

  • Kim, Min-Jung;Chu, Won-Mi;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1041-1048
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    • 2012
  • Shiitake mushroom (SM; Lentinus edodes) are cultivated and consumed in many Asian countries including Vietnam, China, Japan, Korea, and Thailand. In Asia, SM are mainly dried and used as flavoring. The aim of this study was to compare the effects of SM created with different drying processes, such as oven-dried and sun-dried, on the antioxidative and antigenotoxic effects. Raw and dried SM were extracted with acetone, ethanol, methanol, and hot water. The antioxidant effects of SM were evaluated by determining total phenolic content, DPPH radical scavenging activity (RSA), an ORAC assay, and a cellular antioxidant capacity (CAC) assay. The inhibitory effect of SM on oxidative stress-induced DNA damage in human leukocytes was evaluated by a Comet assay. The total phenolic content of raw SM extracted with methanol and of that extracted with water were significantly higher than the dried SM. Among the water extracts, the $IC_{50}$ for DPPH RSA of raw and sun-dried SM were significantly higher than that of oven-dried SM. Sun-dried SM showed the most potent ORAC value at 50 g/mL. The CAC against $AAPH^-$ induced oxidative stress in HepG2 cells, and $H_2O_2$ induced DNA damage were effectively protected against by all SM extracts. These results suggest that unprocessed SM are the best antioxidants, and that the sun-dried method would be the best option to use in terms of antioxidant activity and the antigenotoxic effect.

Generation and Characterization of a Neutralizing Human Monoclonal Antibody to Hepatitis B Virus PreS1 from a Phage-Displayed Human Synthetic Fab Library

  • Jo, Gyunghee;Jeong, Mun Sik;Wi, Jimin;Kim, Doo Hyun;Kim, Sangkyu;Kim, Dain;Yoon, Jun-Yeol;Chae, Heesu;Kim, Kyun-Hwan;Hong, Hyo Jeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1376-1383
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    • 2018
  • The hepatitis B virus (HBV) envelope contains small (S), middle (M), and large (L) proteins. PreS1 of the L protein contains a receptor-binding motif crucial for HBV infection. This motif is highly conserved among 10 HBV genotypes (A-J), making it a potential target for the prevention of HBV infection. In this study, we successfully generated a neutralizing human monoclonal antibody (mAb), 1A8 (IgG1), that recognizes the receptor-binding motif of preS1 using a phage-displayed human synthetic Fab library. Analysis of the antigen-binding activity of 1A8 for different genotypes indicated that it can specifically bind to the preS1 of major HBV genotypes (A-D). Based on Bio-Layer interferometry, the affinity ($K_D$) of 1A8 for the preS1 of genotype C was 3.55 nM. 1A8 immunoprecipitated the hepatitis B virions of genotypes C and D. In an in vitro neutralization assay using HepG2 cells overexpressing the cellular receptor sodium taurocholate cotransporting polypeptide, 1A8 effectively neutralized HBV infection with genotype D. Taken together, the results suggest that 1A8 may neutralize the four HBV genotypes. Considering that genotypes A-D are most prevalent, 1A8 may be a neutralizing human mAb with promising potential in the prevention and treatment of HBV infection.

Protection by Chrysanthemum zawadskii extract from liver damage of mice caused by carbon tetrachloride is maybe mediated by modulation of QR activity

  • Seo, Ji-Yeon;Lim, Soon-Sung;Park, Ji-A;Lim, Ji-Sun;Kim, Hyo-Jung;Kang, Hui-Jung;YoonPark, Jung-Han;Kim, Jong-Sang
    • Nutrition Research and Practice
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    • v.4 no.2
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    • pp.93-98
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    • 2010
  • Our previous study demonstrated that methanolic extract of Chrysanthemum zawadskii Herbich var. latilobum Kitamura (Compositae) has the potential to induce detoxifying enzymes such as NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2) (NQO1, QR) and glutathione S-transferase (GST). In this study we further fractionated methanolic extract of Chrysanthemum zawadskii and investigated the detoxifying enzyme-inducing potential of each fraction. The fraction (CZ-6) shown the highest QR-inducing activity was found to contain (+)-(3S,4S,5R,8S)-(E)-8-acetoxy-4-hydroxy-3-isovaleroyloxy-2-(hexa-2,4-diynyliden)-1,6-dioxaspiro [4,5] decane and increased QR enzyme activity in a dose-dependent manner. Furthermore, CZ-6 fraction caused a dose-dependent enhancement of luciferase activity in HepG2-C8 cells generated by stably transfecting antioxidant response element-luciferase gene construct, suggesting that it induces antioxidant/detoxifying enzymes through antioxidant response element (ARE)-mediated transcriptional activation of the relevant genes. Although CZ-6 fraction failed to induce hepatic QR in mice over the control, it restored QR activity suppressed by $CCl_4$ treatment to the control level. Hepatic injury induced by $CCl_4$ was also slightly protected by pretreatment with CZ-6. In conclusion, although CZ-6 fractionated from methanolic extract of Chrysanthemum zawadskii did not cause a significant QR induction in mice organs such as liver, kidney, and stomach, it showed protective effect from liver damage caused by $CCl_4$.

A Study on the Labeling Efficiency and Cytotoxicity of Hepatocyte-targeting Galactosylated Chitosan Compounds (간세포 지향성 Galactosylated Chitosan 화합물의 표지 수율 향상 및 세포 독성에 대한 연구)

  • Kim, Dae-Weung;Jeong, Hwan-Jeong;Kim, Eun-Mi;Kim, Se-Lim;Kang, Yun-Hee;Kim, Min-Woo;Kim, Chang-Guhn;Sohn, Myung-Hee
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.5
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    • pp.278-283
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    • 2005
  • Purpose: In prior study, we synthesized $^{99m}Tc$-galactosylated chitosan (GC) and performed in vivo biodistribution study, showed specific targeting to hepatocyte. The aim of this study is to evaluate the labeling efficiency and cytotoxicity of modified galactosylated chitosan compounds, galactosyl methylated chitosan (GMC) and HYNIC-galactosylated chitosan (GCH). Materials and Methods: GC, GMC and GCH were synthesized and radiolabeled with $^{99m}Tc$. Then, they were incubated for 6 hours at room temperature and human serum at $37^{\circ}C$. Labeling efficiencies were determined at 15, 30 m, 1, 2, 3 and 6 h after radiolabeling. To evaluate cytotoxicity, MTT assay was performed in HeLa and HepG2 cells. Results: In comparison with them of $^{99m}Tc$-GC labeling efficiencies of $^{99m}Tc$-GMC were significantly improved (100, 97 and 89%) in acetone and 96.3, 95.8 and 75.6% in saline at 15 m, 1 and 6 h, respectively). Moreover, $^{99m}Tc$-GCH showed more improved labeling efficiencies (>95% in acetone and human serum and >90% in saline at 6 h). In MTT assay, cytotoxicity was very low and not different from that of controls. Conclusion: These results represent that these compounds are radiochemically compatible radiopharmaceuticals, can be used in hepatocyte specific imaging study and in vivo gene or drug delivery monitoring.