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Antioxidant Activity and Safety Evaluation of Juice Containing Protaetia brevitarsis (흰점박이꽃무지(Protaetia brevitarsis) 함유 음료의 in vitro 항산화 관련 생리활성효능 및 안전성 검증)

  • Park, Jae-Hee;Kim, So-Yun;Kang, Min-Gu;Yoon, Min-Soo;Lee, Yang-Il;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.1
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    • pp.41-48
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    • 2012
  • The purpose of this study was to assess the antioxidant activity of vegetable extracts (pumpkin, aloe, and artichoke) containing Protaetia brevitarsis (PB) and the clinical and pathological changes in ICR mice after a single oral administration. The total polyphenol (TP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability, total radical trapping antioxidant potential (TRAP), oxygen radical absorbance activity (ORAC), and single cell gel electrophoresis assay were done to measure their antioxidant activities. The effect of vegetable extracts containing PB in TP and the ORAC value was significantly higher than those without PB. In addition, all extracts had effective $DPPH{\cdot}$ scavenging and $ABTS{\cdot}+$ scavenging activities. The protective effect of vegetable extracts with/without PB on $H_2O_2$-induced DNA damage was found. In a single-dose toxicity study, mortality, body weight, physiological signs, and biochemical analysis were analyzed. Seventy mice were randomly assigned to 7 experimental groups and were administered three vegetable extracts with and without PB (2 g/kg). A full 14 days after administration, no mice mortality was observed in any group. Body weight, physiological signs, and biochemical analysis were never significantly different from those of the control group. Taken together, these findings indicate that vegetable extracts containing PB with antioxidant activities and safety could be applied as medicinal and edible resources in an industrial area.

Cytotoxicity and DNA Damage Induced by Magnetic Nanoparticle Silica in L5178Y Cell

  • Kang, Jin-Seok;Yum, Young-Na;Park, Sue-Nie
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.261-266
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    • 2011
  • As recent reports suggest that nanoparticles may penetrate into cell membrane and effect DNA condition, it is necessary to assay possible cytotoxic and genotoxic risk. Three different sizes of magnetic nanoparticle silica (MNP@$SiO_2$) (50, 100 and 200 nm diameter) were tested for cytotoxicity and DNA damage using L5178Y cell. MNP@$SiO_2$ had constant physicochemical characteristics confirmed by transmission electron microscope, electron spin resonance spectrometer and inductively coupled plasma-atomic emission spectrometer for 48 h. Treatment of MNP@$SiO_2$ induced dose and time dependent cytotoxicity. At 6 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$ compared to vehicle control (p<0.05 or p<0.01). Moreover, at 24 h, 50 or 100 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$(p<0.01). And treatment of 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Furthermore, at 48 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Cellular location detected by confocal microscope represented they were existed in cytoplasm, mainly around cell membrane at 2 h after treatment of MNP@$SiO_2$. Treatment of 50 nm MNP@$SiO_2$ significantly increased DNA damage at middle and high dose (p<0.01), and treatment of 100 nm or 200 nm significantly increased DNA damage in all dose compared to control (p<0.01). Taken together, treatment of MNP@$SiO_2$ induced cytotoxicity and enhanced DNA damage in L5178Y cell.

No Relationship between the Amount of DNA Damage and the Level of hMLH1 and RASSF1A Gene Expression in Bladder Cancer Cells Treated with Cisplatin and Gemcitabine

  • de Camargo, Elaine Aparecida;da Silva, Glenda Nicioli;Gobette, Camila Pereira;de Castro Marcondes, Joao Paulo;Salvadori, Daisy Maria Favero
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5941-5948
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    • 2013
  • Tumor response to antineoplastic drugs is not always predictable. This is also true for bladder carcinoma, a highly recurrent neoplasia. Currently, the combination of cisplatin and gemcitabine is well accepted as a standard protocol for treating bladder carcinoma. However, in some cases, this treatment protocol causes harmful side effects. Therefore, we investigated the roles of the genes TP53, RASSF1A (a tumor suppressor gene) and hMLH1 (a gene involved in the mismatch repair pathway) in cell susceptibility to cisplatin/gemcitabine treatment. Two bladder transitional carcinoma cell (TCC) lines, RT4 (wild-type TP53) and 5637 (mutated TP53), were used in this study. First, we evaluated whether the genotoxic potential of cisplatin/gemcitabine was dependent on TP53 status. Then, we evaluated whether the two antineoplastic drugs modulated RASSF1A and hMLH1 expression in the two cell lines. Increased DNA damage was observed in both cell lines after treatment with cisplatin or gemcitabine and with the two drugs simultaneously, as depicted by the comet assay. A lack of RASSF1A expression and hypermethylation of its promoter were observed before and after treatment in both cell lines. On the other hand, hMLH1 downregulation, unrelated to methylation status, was observed in RT4 cells after treatment with cisplatin or with cisplatin and gemcitabine simultaneously (wild-type TP53); in 5637 cells, hMLH1 was upregulated only after treatment with gemcitabine. In conclusion, the three treatment protocols were genotoxic, independent of TP53 status. However, cisplatin was the most effective, causing the highest level of DNA damage in both wild-type and mutated TP53 cells. Gemcitabine was the least genotoxic agent in both cell lines. Furthermore, no relationship was observed between the amount of DNA damage and the level of hMLH1 and RASSF1A expression. Therefore, other alternative pathways might be involved in cisplatin and gemcitabine genotoxicity in these two bladder cancer cell lines.

Neuroprotective Effects of Cheongnoemyeongsin-hwan against Hydrogen Peroxide-induced DNA Damage and Apoptosis in Human Neuronal-Derived SH-SY5Y Cells (인체 신경세포에서 청뇌명신환(淸腦明神丸)의 산화적 스트레스에 대한 세포보호 효과)

  • Pi, Guk Hyun;Hwang, Won Deuk
    • 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.

Lymphocyte DNA Damage and Anti-Oxidative Parameters are Affected by the Glutathione S-Transferase (GST) M1 and T1 Polymorphism and Smoking Status in Korean Young Adults (흡연 여부에 따른 Glutathione S-transferase (GST) M1 및 T1 유전자 다형성이 우리나라 젊은 성인의 임파구 DNA 손상과 항산화 영양상태 지표들 간의 관련성에 미치는 영향)

  • Han, Jeong-Hwa;Lee, Hye-Jin;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.44 no.5
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    • pp.366-377
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    • 2011
  • Glutathione S-transferase (GST) is a multigene family of phase II detoxifying enzymes that metabolize a wide range of exogenous and endogenous electrophilic compounds. GSTM1 and GSTT1 gene polymorphisms may account for inter-individual variability in coping with oxidative stress. We investigated the relationships between the level of lymphocyte DNA and antioxidative parameters and the effect on GST genotypes. GSTM1 and GSTT1 were characterized in 301 young healthy Korean adults and compared with oxidative stress parameters such as the level of lymphocyte DNA, plasma antioxidant vitamins, and erythrocyte antioxidant enzymes in smokers and non smokers. GST genotype, degree of DNA damage in lymphocytes, erythrocyte activities of superoxide dismutase, catalase, and glutathione peroxidase (GSH-Px), and plasma concentrations of total radical-trapping antioxidant potential (TRAP), vitamin C, ${\alpha}$- and ${\gamma}$-tocopherol, ${\alpha}$- and ${\beta}$-carotene, and cryptoxanthin were analyzed. Lymphocyte DNA damage assessed by the comet assay was higher in smokers than that in non-smokers, but the levels of plasma vitamin C, ${\beta}$-carotene, TRAP, erythrocyte catalase, and GSH-Px were lower than those of non-smokers (p < 0.05). Lymphocyte DNA damage was higher in subjects with the GSTM1- or GSTT1-present genotype than those with the GSTM1-present or GSTT1- genotype. No difference in erythrocyte antioxidant enzyme activities, plasma TRAP, or vitamin levels was observed in subjects with the GSTM1 or GSTT1 genotypes, except ${\beta}$-carotene. Significant negative correlations were observed between lymphocyte DNA damage and plasma levels of TRAP and erythrocyte activities of catalase and GSH-Px after adjusting for smoking pack-years. Negative correlations were observed between plasma vitamin C and lymphocyte DNA damage only in individuals with the GSTM1-present or GSTT1- genotype. The interesting finding was the significant positive correlations between lymphocyte DNA damage and plasma levels of ${\alpha}$-carotene, ${\beta}$-carotene, and cryptoxanthin. In conclusion, the GSTM1- and GSTT1-present genotypes as well as smoking aggravated antioxidant status through lymphocyte DNA damage. This finding confirms that GST polymorphisms could be important determinants of antioxidant status in young smoking and non-smoking adults. Consequently, the protective effect of supplemental antioxidants on DNA damage in individuals carrying the GSTM1- or GSTT1-present genotypes might show significantly higher values than expected.

Gender-Specific Changes of Plasma MDA, SOD, and Lymphocyte DNA Damage during High Intensity Exercise (고강도 운동 시 성별에 따른 혈장 MDA, SOD 및 임파구 DNA 손상 변화)

  • Cho, Su-Youn;Chung, Young-Soo;Kwak, Yi-Sub;Roh, Hee-Tae
    • Journal of Life Science
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    • v.21 no.6
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    • pp.838-844
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    • 2011
  • The purpose of this study was to investigate gender-specific changes of plasma MDA, SOD, and lymphocyte DNA damage during high intensity exercise. In this study, 17 healthy male and 18 healthy female college students ran on a treadmill at 85%$VO_{2max}$ until the point of all-out. Blood-collecting was carried out five times (Rest, Ex-Exha, R0.5h, R4h and R24h), and with the collected blood, plasma malondialdehyde (MDA), superoxide dismutase (SOD), and lymphocyte DNA damage were analyzed. Plasma MDA and SOD concentration increased significantly at the Ex-Exha (p<0.05), and there were no significant differences in gender. For the degree of lymphocyte DNA damage, all %DNA in the tail, tail length and tail moment increased significantly at the Ex-Exha (p<0.05), and %DNA in the tail and tail length were significantly higher in the male group than in the female group (p<0.05). These results suggest that acute high intensity exercise not only causes oxidative stress but also brings about lymphocyte DNA damage. In addition, it was found that males showed higher DNA damage than females in terms of oxidative stress subject to high intensity exercise. Nevertheless, further subsequent studies are required in order to better understand the mechanism behind DNA damage varying with gender, in a way that takes into consideration physical fitness, hormonal level, exercise intensity and duration - additional factors which might affect DNA damage.

Antioxidant Activity and DNA Damage Protective Effect of a Robinia pseudoacacia L. Flower Extract (아까시 꽃 추출물의 항산화 활성 및 DNA 손상 억제 효과)

  • Kim, Su-Jung;Seo, Go-Un;Seo, Bo-Young;Park, Eun-Ju;Lee, Seung-Cheol
    • Korean journal of food and cookery science
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    • v.27 no.4
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    • pp.99-106
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    • 2011
  • The antioxidant properties of Robinia pseudoacacia L. water and 70% ethanol extracts were evaluated by determining total phenolic content (TPC), DPPH radical scavenging activity (RSA), and reducing power (RP). The water extract showed higher TPC (9.07 mg/g gallic acid equivalents) and RP than those of ethanol extract, whereas ethanol extract had greater DPPH RSA. The R. pseudoacacia L. extracts also showed antigenotoxic effects for 200 ${\mu}M$ $H_2O_2$-induced DNA damage in human leukocytes. The 200 ${\mu}M$ $H_2O_2$-induced DNA damage decreased following treatment with the water extract. Reductions in DNA damage with 50 ${\mu}g/ml$ of the water and ethanol extracts were 46.5 and 32.4%, respectively.

Changes in the Antioxidative and Antigenotoxic Effects After the Cooking Process of Sulgidduk Containing Pine Needle Juice (솔잎착즙액을 첨가한 설기떡의 가열조리에 따른 항산화 및 항유전독성 효능 검증)

  • Lee, Hyun-Jung;Kim, So-Yun;Park, Jae-Hee;Kim, Rae-Young;Jeong, Hyeon-Suk;Park, Eunju
    • Korean journal of food and cookery science
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    • v.29 no.5
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    • pp.453-462
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    • 2013
  • Recently, two formulas of sulgidduk added to pine needle juice (PNJ) with various physiological activities were developed for metabolic syndrome patients in our lab. According to previous studies, cooking may alter antioxidant properties by initiating destruction, release or transformation of antioxidant compounds contained in food. Therefore, the aim of this study was to compare the antioxidant activities and antigenotixic effects of sulgidduk with/without PNJ and to note changes in these activities after cooking. The ingredients of sulgidduk was added on the basis of 100% rice flour as follows: conventional sulgidduk (S): 1.5% salt, 30.0% sugar; PNJ added to sulgidduk A (PS-A): 1.4% salt, 30.0% sugar, and 1.0% PNJ; PNJ added to sulgidduk B (PS-B): 1.5% salt, 21.4% sugar, and 1.4% PNJ. Ethanol and water extracts of sulgidduk were analyzed for the total phenolic content (TPC), DPPH radical scavenging activity (DPPH RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and antigenotoxic effect by comet assay. The ethanol extracts PS-A and PS-B showed higher TPC and antioxidant activities (DPPH RSA, TRAP, and ORAC) than did the S ethanol extract before cooking. The more PNJ was added, the higher TPC and anitoxidant activities were observed in sulgidduk (PS-A$200{\mu}M$ of $H_2O_2$. Taken together, this study suggests that sulgidduk added to 1.44% of pine needle juice may be a good option antioxidant and antigenotoxic source.

Evaluation of DNA Damage and Repair Kinetics in the Earthworm (Eisenia fetida) Exposed to Radiation and Mercury (방사선과 수은에 의해 유도된 Eisenia fetida 체강세포의 DNA 손상 및 수복 평가)

  • Ryu, Tae-Ho;Nili, Mohammad;An, Kwang-Guk;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.68-73
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    • 2011
  • The single cell gel electrophoresis (SCGE) assay is a microelectrophoretic technique for assessments of DNA damage at the level of the individual eukaryotic cell. The SCGE assay, due to its simplicity, sensitivity and need of a few cells, has advantages compared to other genomic damage assays such as sister chromatid exchange, chromosomal aberration and micronucleus test. In this study, investigated were the levels of DNA damage and the repair kinetics in the coelomocytes of Eisenia fetida treated with HgCl2 and ionizing radiation by means of the SCGE assay. For detecting DNA damage and repair in coelomocytes, earthworms (E. fetida) were irradiated with six doses of ${\gamma}$-rays (0, 2.5, 5, 10, 20 and 50 Gy) and in vivo exposed to mercuric chloride at 0, 80 and 160 mg $kg^{-1}$ for 48 hours. Then the Olive tail moments were measured during 0~12 hours after irradiation and 0~72 hours after Hg treatment. The results showed that the more the oxidative stress was induced by mercury and radiation, the longer the repair time was required. Also, the results suggest that the SCGE assay may be used as an important tool for comparison of the sensitivity of different species to oxidative stresses.

Antioxidative Activity of Extracts from Wisteria floribunda Flowers (등나무 꽃 추출물의 항산화 활성)

  • Oh, Won-Gyeong;Jang, In-Cheol;Jeon, Gyeong-Im;Park, Eun-Ju;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.677-683
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    • 2008
  • The antioxidant activities of Wisteria floribunda flowers (WFF) were evaluated. The samples were prepared by extracting separately two different colored flowers (purple and white) with four different solvents (methanol, ethanol, acetone, and water). The antioxidant properties were evaluated by determining total phenolic contents (TPC), radical scavenging activity (RSA), and reducing power (RP). Water extract from purple WFF and ethanol extract of white WFF showed the highest total phenol contents (491 and 787 ${\mu}M$ gallic acid equivalents), respectively. Water extracts of purple and white WFF also showed higher RSA. In the case of RP, ethanol extract of purple WFF, methanol and water extracts of white WFF showed relatively higher values. The 200 ${\mu}M$ $H_2O_2$ induced oxidative DNA damage in human leukocytes was significantly inhibited with WFF extracts excluding ethanol and acetone extracts of purple flowers. These results suggest that W. floribunda flowers have significant antioxidative activity and protective effect against oxidative DNA damage.