• Title/Summary/Keyword: effect on DNA

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Antioxidative and Probiotic Properties of Lactobacillus gasseri NLRI-312 Isolated from Korean Infant Feces

  • Kim, H.S.;Jeong, S.G.;Ham, J.S.;Chae, H.S.;Lee, J.M.;Ahn, C.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1335-1341
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    • 2006
  • We selected a Lactobacillus spp. from Korean healthy infant feces based upon their antioxidant activity. This strain was identified as Lactobacillus gasseri by 16S rDNA sequencing, and named Lactobacillus gasseri NLRI-312. In the present study, we investigate the protective effect of this strain on the $H_2O_2$ induced damage to cellular membrane lipid and DNA in Jurkat cells. To estimate the extent of cellular lipid peroxidation inhibition, MDA (malondialdehyde) was measured, and DNA damage was tested by the comet assay. We also examined probiotic properties including tolerance to acid and bile, antibiotic resistance. From the results obtained, the supplementation of Jurkat cells with NLRI-312 decreased in DNA damage, while no effect was shown on MDA decrease. In probiotic properties, this strain was resistance to both acid and bile, showed considerably higher survival when incubated in pH 2 or 1% bile salts (w/v). We concluded that the NLRI-312 could be used as potential probiotic bacteria, with the effect of reducing DNA damage induced by $H_2O_2$.

Protective Effects of Ginkgo Biloba Leaf Extract(GBE) against 1,2,4-benzenetriol Induced Toxicity in Vitro (Ginkgo biloga 잎 추출물의 1,2,4-benzenetriol에 대한 항산화 효과에 대한 연구)

  • 이영준;김태연;정해원
    • Journal of Environmental Health Sciences
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    • v.27 no.1
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    • pp.124-130
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    • 2001
  • Ginkgo biliba has been used for bronchitis and asthma in oriental countries and its leaf extract(GBE) contains 24% ginkgoflavone glycoside and 6% terpenoid. Flavonoids and terpenoids are known to have various antioxidant effects such as scavenging of free radicals and chelation of transtional metals. Antioxidant effect of GBE against 1,2,4-benzenetriol(BT), one of toxic metabolites of benzene, was demonstrated throughbsister chromatid exchange(SCE) analysis, single cell gel electrophoresis(SCGE) analysis, DNA cleavage assay and lipid peroxidation production analysis. The means of SCE frequencies at 10, 25 and 50$\mu$M concentration of BT were 7.72, 8.02, 9.22 respectively. In addition of GBE with concentration of 50, 200 and 500$\mu\textrm{g}$/$m\ell$, SCE frequencies were decreased significantly.(p<0.05) According to SCGE analysis, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 $\mu$m and the DNA damage induced by BT was significantly protected by GBE(p<0.001). No genotoxicity was observed by GBE treatment alone on DNA cleavage. The effect of BT on lipid peroxidation product, Malondiadehyde(MDA), was increased with concentration of BT(10 and 50 $\mu$M) and reduction in MDA was noted when GBE was added. From above results it is suggested that GBE could protect the cell and DNA from pro-oxidant effect by reactive oxigen species induced by BT.

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Non-histone protein HMGB1 inhibits the repair of damaged DNA by cisplatin in NIH-3T3 murine fibroblasts

  • Yusein-Myashkova, Shazie;Ugrinova, Iva;Pasheva, Evdokia
    • BMB Reports
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    • v.49 no.2
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    • pp.99-104
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    • 2016
  • The nuclear non-histone protein high mobility group box (HMGB) 1 is known to having an inhibitory effect on the repair of DNA damaged by the antitumor drug cisplatin in vitro. To investigate the role of HMGB1 in living cells, we studied the DNA repair of cisplatin damages in mouse fibroblast cell line, NIH-3T3. We evaluated the effect of the post-synthetic acetylation and C-terminal domain of the protein by overexpression of the parental and mutant GFP fused forms of HMGB1. The results revealed that HMGB1 had also an inhibitory effect on the repair of cisplatin damaged DNA in vivo. The silencing of HMGB1 in NIH-3T3 cells increased the cellular DNA repair potential. The increased levels of repair synthesis could be "rescued" and returned to less than normal levels if the knockdown cells were transfected with plasmids encoding HMGB1 and HMGB1 K2A. In this case, the truncated form of HMGB1 also exhibited a slight inhibitory effect.

Protective Effect of Flavonoids on Lymphocyte DNA Damage Using Comet Assay (Comet Assay를 이용한 Flavonoids와 항산화 비타민의 인체임파구 세포 DNA 손상 보호 효과)

  • 박유경;전은재;강명희
    • Journal of Nutrition and Health
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    • v.36 no.2
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    • pp.125-132
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    • 2003
  • The present study was attempted to investigate and compare the antioxidant potency of several well-know flavonoids, antioxidant vitamin and commercially available popular beverages. The antioxidant potency was assessed by the effect on reducing oxidative DNA damage of human lymphocytes. Cellular oxidative DNA damage was measured by SCGE (single-cell gel electrophoresis), also known as comet assay. Lymphocytes were pre-treated for 30 minutes with wide ranges of doses of apigenin, kaempferol, luteolin, myricetin, rutin, quercetin, $\alpha$-tocopherol (10,25,50,100,200,500,1000 $\mu$M) ,green tea extract or grape juice (10,50,100,250,500,1000 $\mu$g/mL) followed by a $H_2O$$_2$(100 $\mu$M) treatment for 5 min as an oxidative stimulus. The physiological function of each antioxidant substance on oxidative DNA damage was analyzed as tail moment (tail length $\times$ percentage migrated DNA in tail) and expressed as relative DNA damage score after adjusting by the level of control treatment. Cells treated with $H_2O$$_2$alone (positive control) had an extensive DNA damage compared with cells treated with phosphate buffered saline (PBS, negative control) or pre-treated with all the tested samples. Of all the six flavonoids, quercetin was the most potent antioxidant showing the lowest $ED_{50}$/ of 8.5 $\mu$g/mL (concentration to produce 50% protection of relative DNA damage). The antoxidant potency of individual flavonoids were ranked as follows in a decreasing order; luteolin (18.4 $\mu$g/mL), myricetin (19.0 $\mu$g/mL) , rutin (22.2 $\mu$g/mL) , apigenin (24,3 $\mu$g/mL) , kaempferol (25.5 $\mu$g/mL). The protective effect of $\alpha$-tocopherol was substantially lower (highest $ED_{50}$value of 55.0 $\mu$g/mL) than all the other flavonoids, while the protective effect was highest in green tea and grape juice with low ED5O value of 7.6 and 5.3, respectively. These results suggest that flavonoids, especially quercetin, and natural compounds from food product, green tea and grape juice, produced powerful anti-oxidative activities, even stronger than $\alpha$-tocopherol. Taken together, supplementation of antioxidants to lymphocytes followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.

Effect of Drug Loading on the Physicochemical Properties and Stability of Cationic Lipid-based Plasmid DNA Complexes

  • Jeong, Ui-Hyeon;Jung, Ji-Hye;Davaa, Enkhzaya;Park, Se-Jin;Myung, Chang-Seon;Park, Jeong-Sook
    • Journal of Pharmaceutical Investigation
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    • v.39 no.5
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    • pp.339-343
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    • 2009
  • Recently, co-delivery of drug and gene has been attempted for higher therapeutic effects of anticancer agents. In this study, cationic liposomes were prepared using 1,2-dioleoyl-3-trimethylammoniopropane (DOTAP) as a cationic lipid to investigate the effect of drug loading on the physicochemical characteristics of cationic liposomes/DNA complexes. The complex formation between cationic liposomes and negatively charged plasmid DNA was confirmed and the protection from DNase was observed. Particle size of complexes was reduced not by drug loading, but by the increased ratio of cationic lipid to plasmid DNA. Meanwhile, zeta potential of complex was increased by the addition of cationic liposomes to complexes and the effect of drug loading on the zeta potential was not much higher than on particle size. Gel retardation of complexes was indicated when the complexation weight ratios of cationic lipid to plasmid DNA were higher than 24:1 for drug free complexes and 20:1 for drug loaded ones, respectively. Agarose gel retardation showed the similar complexation between plasmid DNA and drug free liposomes or drug loaded liposomes. Both complexes protected plasmid DNA from DNase independent of complexation temperature. From the results, drug loading may affect not the complex formation of cationic liposomes and plasmid DNA, but the particle size of complex.

Effects of Aronia melanocarpa and Korean Red Ginseng Ethanol Extracts Combination on Cytotoxicity induced by Fludarabine, a DNA Chain Terminating Anti-Cancer Drug (DNA 사슬 종결형 항암제인 플루다라빈에 의해 유도된 세포독성에 대한 아로니아-홍삼 에탄올 혼합 추출물의 효과)

  • Kim, Min Seob;Chung, You Heon;Oh, Hong Geun;Park, Jong Kun
    • The Korean Journal of Food And Nutrition
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    • v.30 no.4
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    • pp.673-680
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    • 2017
  • Fludarabine, a chain terminating anti-cancer drug, is a purine analogue that causes DNA strand breaks in normal cells. In this study, we determined if A. melanocarpa and Korean red ginseng extract mixture reduce cytotoxicity of fludarabine. Treatment of HaCaT cells with $10{\mu}M$ of fludarabine for 24 hours decreased cell viability and increased DNA strand breaks. Treatment of A. melanocarpa and Korean red ginseng extract mixture for 24 hours increased cell viability as compared with single extract treatment. The protective effect of these extracts on cell activity increased in a concentration-dependent manner. DNA strand breaks induced by fludarabine decreased as concentration of extract mixture increased. p-H2AX level, a marker of DNA strand breakage, decreased depending on the concentration of extract mixture. The effect of mixed extract of A. melanocarpa and Korean red ginseng on DNA damage is due to the anti-oxidative effect of A. melanocarpa and signal transmission through glucocorticoid receptor upon binding of saponin of Korean red ginseng.

Antioxidative Activities and Inhibition of DNA Damage of Ethylacetate Fraction from Sophorae Flos and Sophorae fructus (괴화와 괴각 에틸아세테이트 분획물의 항산화 및 산화적 DNA 손상 억제 활성)

  • Jang, Tae Won;Kim, Ye Rang;Lee, Sung Hyeon;Kim, Do-Wan;Park, Jae Ho
    • The Korea Journal of Herbology
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    • v.30 no.2
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    • pp.31-36
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    • 2015
  • Objectives : In this study, we demonstrated the antioxidant activities and the inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Sophorae Flos and Sophorae fructus. Methods : Sophorae Flos and Sophorae fructus were extracted with methanol(MeOH) and divided to Petroleum ether, Ethyl acetate(EtOAC) and Water fraction. The antioxidant activities were conducted by the 1,1-diphenyl-2-picryl hydrazyl(DPPH) radical, 2, 2'-Azine-bis(3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt(ABTS) radical scavenging assay, $Fe^{2+}$ chelating assay and Reducing power assay. The inhibitory effect of DNA damage were characterized on ${\varphi}$ X-174 RF I plasmid DNA cleavage assay. In addition, we analyzed the Total phenol contents and the Vitamin C contents of Sophorae Flos and Sophorae fructus. Results : The results of DPPH were 92.71% and 94.72%, ABTS were 87.16% and 62.44%, and $Fe^{2+}$ chelating were 95.81% and 85.11% at $200{\mu}g/m{\ell}$ of Sophorae Flos and Sophorae fructus respectively. The Sophorae Flos showed stronger effect than Sophorae fructus in Reducing Power assay. Total phenol content was 111.77 mg/g and 122.54 mg/g, and Vitamin C content was 2.59 mg/g and 3.03 mg/g. Also both Sophorae Flos and Sophorae fructus have inhibitory antioxidant effect on ${\varphi}$ X-174 RF I plasmid DNA cleavage assay. Conclusions : Over all, this study suggests that Sophorae Flos and Sophorae fructus can be used as not only effective antioxidant but also natural medicine.

Effect of DNA methylation on the reactivity of DNA alkylating agents

  • Yoo, Ja-Kyung;Park, Hyun-Ju
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.359.1-359.1
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    • 2002
  • In mammalian species, CpG dinucleotides are highly methylated with 60-90% methylation at the 5-position of cytosine. The pattern of DNA methylation in a cell dramatically affects the function of the DNA by switching genes on or off. Abnormal methylation events occur during aging and in the development of many cancers. Methylated CpG was reported recently to affect the reactivity of agents (mitomycin C and benzo [a]pyrenediolepoxide) that can fromguanine adducts in DNA. (omitted)

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Comparison of the Protective Effect of Antioxidant Vitamins and Fruits or Vegetable Juices on DNA Damage in Human Lymphocyte Cells Using the Comet Assay (Comet Assay를 이용한 항산화 비타민과 과일.야채즙의 인체 임파구 세포 DNA 손상 감소 효과 비교)

  • 전은재;박유경;김정신;강명희
    • Journal of Nutrition and Health
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    • v.37 no.6
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    • pp.440-447
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    • 2004
  • In this study the in vitro protective effects of several antioxidant vitamins (vitamin C, $\alpha$-tocopherol, $\beta$-carotene), fruits and vegetables (strawberry, tangerine, orange and 100% orange juice, carrot juice), on the levels of isolated human lymphocyte DNA damage was measured using Comet assay. Comet assay has been used widely to assess the level of the DNA damage in the individual cells. Lymphocytes were pre-treated for 30 minutes with antioxidant vitamins (10, 50, 100, 500 $\mu$M) or fruits$.$vegetables (10, 100, 500, 1000 $\mu$g/ml), an4 then oxidatively challenged with 100 $\mu$M $H_2O$$_2$ for 5 min at 4$^{\circ}C$. The protective effect of antioxidant vitamins against DNA damage at a concentration of 50 $\mu$M were 50% in vitamin C, 32% in $\alpha$-tocopherol, whereas, fJ-carotene showed a 55% protection at a dose as low as 10 $\mu$M. The inhibitory effects of DNA damage by strawberry, tangerine, orange, orange juices, carrot juices were 50 - 60% with wide ranges of doses. The results of the present study indicate that most the antioxidant vitamins and fruits$.$vegetables juices produced a significant reduction in oxidative DNA damage.

Onion Supplementation Inhibits Lipid Peroxidation and Leukocyte DNA Damage due to Oxidative Stress in High Fat-cholesterol Fed Male Rats

  • Park, Jae-Hee;Seo, Bo-Young;Lee, Kyung-Hea;Park, Eun-Ju
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.179-184
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    • 2009
  • The aim of this study was to investigate effects of onion, red onion, or quercetin on plasma antioxidant vitamin, lipid peroxidation, and leukocyte DNA damage in rats fed a high fat-cholesterol diet. Forty SD male rats were assigned to normal control, high fat-cholesterol diet (HF), or HF+5% onion powder, HF+5% red onion powder, or HF+0.0l% quercetin. The HF diet resulted in significantly higher plasma lipid peroxidation which decreased with onion, red onion, or quercetin supplementation. Leukocyte DNA damage induced by HF diet decreased significantly in rats fed onion and red onion, while quercetin supplementation had no effect on preventing leukocyte DNA damage. $H_2O_2$ induced leukocyte DNA damage exhibited a highly significant negative correlation with plasma retinol and tocopherols. These results suggest that onion or red onion powder exerts a protective effect with regard to DNA damage in rats fed HF diet. However, 0.01% quercetin in pure form might not be effective at preventing DNA damage.