• Title/Summary/Keyword: Oxidative damage

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Evaluation of DNA Damage by Mercury Chloride (II) and Ionizing Radiation in HeLa Cells (이온화 방사선 및 염화수은(II)에 의한 자궁경부암 세포의 DNA 손상 평가)

  • Woo Hyun-Jung;Kim Ji-Hyang;Antonina Cebulska-Wasilewska;Kim Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.24 no.1 s.61
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    • pp.46-52
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    • 2006
  • The mercury is among the most highly bioconcentrated toxic trace metals. Many national and international agencies and organisations have targeted mercury for the possible emission control. The mercury toxicity depends on its chemical form, among which alkylmercury compounds are the most toxic. A human cervix uterus cancer cell line HeLa cells was employed to investigate the effect of the toxic heavy metal mercury (Hg) and ionizing radiation. In the in vitro comet assays for the genotoxicity in the HeLa cells, the group of Hg treatment after irradiation showed higher DNA breakage than the other groups. The tail extent moment and olive tail moment of the control group were $4.88{\pm}1.00\;and\;3.50{\pm}0.52$ while the values of the only Hg treatment group were $26.90{\pm}2.67\;and\;13.16{\pm}1.82$, respectively. The tail extent moment and olive tail moment of the only 0.001, 0.005, 0.01 Hg group were $12.24{\pm}1.82,\;8.20{\pm}2.15,\;20.30{\pm}1.30,\;12.26{\pm}0.52,\;40.65{\pm}2.94\;and \;20.38{\pm}1.49$, respectively. In the case of Hg treatment after irradiation, the tail extent moment and olive tail moment of the 0.001, 0.005, 0.01 Hg group were $56.50{\pm}3.93,\;32.69{\pm}2.48,\;62.03{\pm}5.14,\;31.56{\pm}1.97,\;72.73{\pm}3.70\;and \;39.44{\pm}3.23$, respectively. The results showed that Hg induced DNA single-strand breaks or alkali labile sites as assessed by the Comet assay. It is in good agreement with the reported results. The mercury inhibits the repair of DNA. The bacterial formamidopyrimidine-DNA glycosylase (Epg protein) recognizes and removes some oxidative DNA base modifications. Enzyme inactivation by Hg (II) may therefore be due either to interactions with rysteine residues outside the metal binding domain or to very high-affinity binding of Hg (II) which readily removes Zn (II) from the zinc finger.

Antioxidant and Cell Proliferation Effects of Acanthopanax senticosus for Extract in Human Osteoblast-like MG-63 Cell Line (가시오가피 추출물의 항산화활성 및 MG-63 조골세포 증식과 alkaline phosphatase 활성에 미치는 효과)

  • Lim, So-Young;Leem, Jae-Yoon;Lee, Choong-Soo;Jang, Yu-Jung;Park, Jeong-Woo;Yoon, Sun
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.694-700
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    • 2007
  • Acanthopanax senticosus is a common Asian herb also known as "Siberian Ginseng". It is often used as a traditional herbal medicine for reducing damage in the liver, kidney, bone and muscle. In the present study we investigated the ferric reducing/antioxidant power and total phenolic contents of the ethanol-/water-extracts obtained from the stems and leaves of Acanthopanax senticosus. Osteoblast cellular proliferation was evaluated using the MTT and alkaline phosphatase activity assays in the human osteoblast-like MG-63 cell line. Acanthopanax senticosus extracts exerted remarkable ferric reducing/antioxidant power and contained high amount of phenolics. Among the extracts the stem-/ethanol-extract showed the highest antioxidant activity and total phenol content. Interestingly a highly positive correlation was found between antioxidant activity and total phenol content (p < 0.01). Proliferation of MG-63 osteoblast cells was highest in the stem-/ethanol-extract and alkaline phosphatase activity significantly increased in the water-extract of the stems (p < 0.05). These findings suggest that Acanthopanax senticosus extracts have antioxidant activity for preventing oxidative stress-related diseases and may have beneficial effects on bone health through the proliferation of osteoblast cells.

The effect of Allium sativum L. extract on hepatic function in rats with CCl4-induced (hepatic) injury (마늘 열수추출물의 CCl4 유발된 간 손상 랫드에서 기능학적 변화에 관한 연구)

  • Lee, Tae-Jong;Min, Kyung-Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.5
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    • pp.1936-1942
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    • 2010
  • To examine the effect of overdosed functional food in liver-injured patients, we tried to investigate the dose effect of Allium sativum L. (also called garlic) extract on rats with CCl4-induced liver damage, by comparatively examinating the hepatic function of each group. 42 male Sprague-Dawley rats were selected and divided equally (n=7) into normal, control, positive control, and three experimental subgroups (E1, E2 and E3), respectively. For each kg of body weight, hepatic injury was induced by intraperitoneal administration of $0.5\;m{\ell}$ (0.20 g/kg/day) $CCl_4$, which was diluted in equal amount of olive oil, once a day on every other day for total 5 times. Hot water extraction was performed using domestically cultivated organic garlics, and the obtained extract was injected by sonde, once daily during 4 weeks, to each experimental subgroups by different dosage of 0.35 g/kg(E1), 0.70 g/kg(E2), 1.40 g/kg(E3), respectively. Results showed that the injection of garlic extract positively influenced the physiological activation which lowered the oxidative stress, changed the toxicity, and functionally improved the hepatic condition. Comparing the dose effect between the three experimental subgroups (E1, E2, E3), results of the maximal-dosed subgroup (E3) showed less significance compared with the lower-dosed subgroups(E1 and E2), which seems to resulted from the (increased) toxicity of garlic.

Effect of Whey Protein Isolate and Lactobacillus spp. Cell Extracts on Intracellular Antioxidative Activities in Human Prostate Epitherial Cells (유청단백질 및 Lactobacillus spp. 추출물이 전립선 세포 내 항산화 활성에 미치는 영향)

  • 변정열;윤영호
    • Journal of Animal Science and Technology
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    • v.48 no.5
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    • pp.719-726
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    • 2006
  • Bovine whey protein are rich in cysteine, which is the rate limiting amino acid for synthesis of antioxidant glutathione(GSH). Some strains of Lactobacillus caseihas been reported to contain high level of GSH in cell extracts. The objective ofthis study was to determine whether enzymatically hydrolyzed whey protein isolate(WPI) and cell extract of Lb. casei HY2782 could increase intracellular GSH concentrations and protect against oxidant induced cell death in human prostate epithelial cell line (designated as RWPE1, and PC3MMM2 cells). Treatment of RWPE1 cellsandPC3MMM2 cells with hydrolyzed WPI (500g/ml) significantly increased GSH by28.2% and38.4% respectively. Compared with control cells receiving no hydrolyzed WPI(P<0.05). hydrolyzed WPI and Lb casei HY2782 cell extracts significantly protected RWPE1 and PC3MMM2 cellsfrom oxidant induced cell death compared with controls receiving no WPI. DNA damage associated with oxidant treatment was demonstrated by single cell gel (SCG) electrophoresis.

The Effects of Aflatoxin $B_1$Co-administrated with Antioxidant Vitamins on Lipid Contents and Fatty Acids Composition of Liver in Mice (항산화 비타민과 Aflatoxin$B_1$의 혼합 투여가 마우스간의 지질 함량 및 지방산 조성에 미치는 영향)

  • 박선자;박정현;강말순;정덕화
    • Journal of Food Hygiene and Safety
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    • v.15 no.1
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    • pp.5-12
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    • 2000
  • Lipid peroxidation is one of the main manifestations of oxidative damage and has been found play an important role in the toxicity and carcinogenesis of many carcinogens. This study was carried out to investigate the effects of aflatoxin B$_1$co-administrated with antioxidant vitamins on lipid contents and fatty acids components of liver in mice. For this work, vita-min C and vitamin E, the major antioxidants, were administrated with 10 mg/kg and 63.8 mg/kg respectively, through intraperitoneal(i.p) injection to male ICR mice, and 0.4 mg/kg of the AFB$_1$injected by i.p. 1hr later. The results were as follows: two fold amounts of free cholesterol, triglyceride, and total cholesterol in serum and liver of mice treated with only AFB$_1$were observed, when compared to those of mice co-administrated with antioxidant vitamins. However, the levels of phospholipids in serum and liver of mice treated with only AFB$_1$were decreased. Concerning to fatty acids composition of liver from AFB$_1$-treated mice, P/S ratio was shown more low level in cholesteryl ester, triglyceride, total cholesterol and phospholipid than those of mice co-administrated with antioxidant vitamins. In these data which provide with a reliable evidence on their antioxidantal effects to aflatoxicosis.

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Functionality Analysis of Rhus javanica Fermented by Lactobacillus spp. (Lactobacillus spp. 이용 발효 붉나무의 기능성물질 검색에 대한 연구)

  • Lee, Dong-Sung;Kang, Min-Su;Kim, Youn-Chul;Im, Nam-Kyung;Kim, Hyun-Su;Jeong, Gil-Saeng
    • Journal of Life Science
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    • v.23 no.1
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    • pp.44-54
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    • 2013
  • To determine the potential function of Rhus javanica in Korean medicine, it was fermented with each strain of Lactobacillus spp. Each strain of Lactobacillus spp. was inoculated in lactobacilli MRS broth, and 5 mg/ml of methanol extract of Rhus javanica was added. In mouse hippocampal HT22 cells, ethyl acetate extract of R. javanica fermented with L. brevis KCTC 3498 induced heme oxygenase-1 expression and showed a significant cytoprotective effect on glutamate-induced oxidative damage. The cytoprotective effect was related to the transcription of the nuclear factor E2-related factor2 (Nrf2), which is responsible for the induction of heme oxygenase-1 within the nucleus. The antimicrobial, antioxidant, and heme oxygenase-1 expression activities of fermented R. javanica were measured after extraction with ethyl acetate. R. javanica fermented with L. plantarum subsp. plantarum KCTC 3108, L. fermentum KCTC 3112, and L. brevis KCTC 3498 had higher antioxidant activity than nonfermented R. javanica. The fermented R. javanica with L. plantarum subsp. plantarum KCTC 3108, L. casei KCTC 3109 after ethyl acetate extraction showed antibacterial activity against Bacillus subtilis PCI 219, Escherichia coli KCTC 1682, Shigella flexneri KCTC 2517, Vibrio parahaemolyticus KCTC 7471, and Pseudomonas aeruginosa KCTC 2004. An ethyl acetate extract of the fermented R. javanica with Lactobacillus brevis KCTC 3498 exhibited stronger antibacterial activity than a nonfermented one against strains of B. subtilis PCI 219, E. coli KCTC 1682, S. flexneri KCTC 2517, and V. parahaemolyticus KCTC 7471.

Effects of Tumor Microenvironmental Factors on DNA Methylation and Radiation Sensitivity in A549 Human Lung Adenocarcinoma

  • Oh, Jung-Min;Kim, Young-Eun;Hong, Beom-Ju;Bok, Seoyeon;Jeon, Seong-Uk;Lee, Chan-Ju;Park, Dong-Young;Kim, Il Han;Kim, Hak Jae;Ahn, G-One
    • Journal of Radiation Protection and Research
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    • v.43 no.2
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    • pp.66-74
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    • 2018
  • Background: Tumor response to anticancer therapies can much be influenced by microenvironmental factors. In this study, we determined the effect of these microenvironmental factors on DNA methylation using A549 human lung adenocarcinoma cell line. Materials and Methods: We subjected A549 cells to various conditions mimicking tumor microenvironment including hypoxia, acidosis (sodium lactate), oxidative stress ($H_2O_2$), bystander effect (supernatant from doxorubicin (Dox)-treated or irradiated cells), and immune cell infiltration (supernatant from THP-1 or Jurkat T cells). Genomic DNA was isolated from these cells and analyzed for DNA methylation. Clonogenic cell survival, gene expression, and metabolism were analyzed in cells treated with some of these conditions. Results and Discussion: We found that DNA methylation level was significantly decreased in A549 cells treated with conditioned media from Dox-treated cells or Jurkat T cells, or sodium lactate, indicating an active transcription. To determine whether the decreased DNA methylation affects radiation sensitivity, we exposed cells to these conditions followed by 6 Gy irradiation and found that cell survival was significantly increased by sodium lactate while it was decreased by conditioned media from Dox-treated cells. We further observed that cells treated with conditioned media from Dox-treated cells exhibited significant changes in expression of genes including BAX and FAS (involved in apoptosis), NADPH dehydrogenase (mitochondria), EGFR (cellular survival) and RAD51 (DNA damage repair) while sodium lactate increased cellular metabolism rather than changing the gene expression. Conclusion: Our results suggest that various tumor microenvironmental factors can differentially influence DNA methylation and hence radiosensitivity and gene expression in A549 cancer cells.

Induction of Metallothionein Gene by Laminin in Normal and Malignant Human Prostate Epithelial Cells (악성 단계별 인간 전립선 암세포에서 라미닌에 의한 metallothionein 유전자 발현유도 현상 연구)

  • Ock, Mee-Sun;Cha, Hee-Jae
    • Journal of Life Science
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    • v.21 no.4
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    • pp.529-533
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    • 2011
  • Metallothioneins (MT) are a group of low-molecular weight, cysteine-rich, intracellular proteins that are encoded by a family of genes containing at least 10 functional isoforms in human. The expression and induction of these proteins is associated with protection against DNA damage, oxidative stress, and apoptosis. Many studies have shown increased expression of MT in various human tumors, whereas MT is down-regulated in certain tumors such as hepatocellular carcinoma and liver adenocarcinoma. Hence, the expression of MT is not universal to all human tumors but may depend on the differentiation status and proliferative index of tumors, along with other tissue factors and gene mutations. Using Northern blot analysis, we found that laminin induced expression of MT-1 in HSG and PC12 cells, which can be differentiated by laminin, but had no effect on MB-231, MDA-435, and PC-3 cells, which cannot be differentiated by laminin. In addition, we analyzed the expression level of the MT-1 gene in five prostate cancer cell lines possessing different metastatic potential. The expression of MT-1 in normal and less malignant cells (RWPE-1 and WPE1-NA22) was high and up-regulated by laminin, whereas the expression of MT-1 in WPE1-NB14, WPE1-NB11, and WPE1-NB26 cells (malignant) was extremely low and not elevated by laminin. These results suggest that the MT-1 gene is involved in laminin-mediated differentiation and affects the metastatic potential of tumor cells.

Antioxidative Activity of Cornus officianalis Extracts Obtained by Four Different Extraction Techniques (산수유 추출방법에 따른 항산화 기능 분석)

  • Park, Eun-Bi;Kim, Hye-Sun;Shin, So-Yun;Ji, In-Ae;Kim, Ji-Hyun;Kim, Sung-Goo;Yoo, Byung Hong;Kim, Byung-Woo;Kwak, Inseok;Kim, Moon-Moo;Chung, Kyung Tae
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1507-1514
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    • 2012
  • Oxidative stress leads to damage in all components of the cell, including proteins, lipids, and DNA due to imbalance between reactive oxygen species production and cellular detoxification. Phytochemicals are well-known to contain antioxidants, and their physiological role has been intensively studied. The fruit of Cornus officianalis has been used in oriental medicine and has been reported to have many functions. In this study, four different extraction techniques were applied to extract functional components from the fruit of Cornus officianalis, and the content of loganin, which is an antioxidant having DPPH radical and hydrogen peroxide scavenging activity and reducing power, was analyzed in each extract. Extraction techniques employed in this study were heat extraction by water, 70% ethanol extraction, enzyme treatment, and combination of enzyme treatment and heat extraction by water. All extracts contained 11.8-18.0 mg/g loganin and showed antioxidation function assayed by measuring DPPH radical and hydrogen peroxide scavenging activity and reducing power. Among them, heat extraction was the most effective technique, showing a maximum amount of loganin (18.0 mg/g) and antioxidative activity at 100 mg/ml concentration. Each extract showed very low cytotoxicity up to at 500 mg/ml but 10-20% cytotoxicity at 1,000 mg/ml by in vitro MTT assay.

Effect of Cordycepin-increased Cordyceps militaris Powder on Tissues Lipid Peroxidation and Antioxidative Activity in Carbon Tetrachloride-induced Hepatic Damage in Rats (Cordycepin이 사염화탄소 유발 간손상 흰쥐의 조직 과산화 지질 농도 및 항산화 활성에 미치는 영향)

  • Ahn, Hee-Young;Park, Kyu-Rim;Kim, Yu-Ra;Cha, Jae-Young;Cho, Young-Su
    • Journal of Life Science
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    • v.23 no.7
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    • pp.904-912
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    • 2013
  • This study aimed to evaluate the protective effect of cordycepin-increased Cordyceps militaris strain on carbon tetrachloride ($CCl_4$)-induced hepatotoxicity and oxidative stress in rats. Male Sprague-Dawley rats were randomly divided into five groups (n=6) based on six dietary categories: normal (N), $CCl_4$ control (C), $CCl_4$ plus Paecilomyces japonica (CPJ) (3%, w/w), $CCl_4$ plus C. militaris (CCM) (3%, w/w), and $CCl_4$ plus cordycepin-increased C. militaris ($CCM{\alpha}$) (3%, w/w). The activities of the liver marker enzymes ALT, AST, and LDH and the levels of lipid peroxidation were increased in the $CCl_4$-treated groups, but these parameters were significantly decreased in the $CCM{\alpha}$ group. The TBARS content in the liver homogenate, microsome, and mitochondrial fractions of the C group was significantly elevated compared with the N group. However, in the $CCl_4$-treated groups, $CCM{\alpha}$ group was significantly lowered in the TBARS levels of hepatic homogenate and microsomal fractions. The C group showed a significant decrease in the levels of plasma and hepatic glutathione, whereas they were significantly increased in the $CCM{\alpha}$ group. Accordingly, cordycepin-increased C. militaris may be an ideal animal model for studying hepatoprotective effects.