• Biomolecules & Therapeutics
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• 제29권1호
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• pp.90-97
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• 2021

Ultraviolet B (UVB) radiation causes DNA base modifications. One of these changes leads to the generation of 8-oxoguanine (8-oxoG) due to oxidative stress. In human skin, this modification may induce sunburn, inflammation, and aging and may ultimately result in cancer. We investigated whether phloroglucinol (1,3,5-trihydroxybenzene), by enhancing the expression and activity of 8-oxoG DNA glycosylase 1 (Ogg1), had an effect on the capacity of UVB-exposed human HaCaT keratinocytes to repair oxidative DNA damage. Here, the effects of phloroglucinol were investigated using a luciferase activity assay, reverse transcription-polymerase chain reactions, western blot analysis, and a chromatin immunoprecipitation assay. Phloroglucinol restored Ogg1 activity and decreased the formation of 8-oxoG in UVB-exposed cells. Moreover, phloroglucinol increased Ogg1 transcription and protein expression, counteracting the UVB-induced reduction in Ogg1 levels. Phloroglucinol also enhanced the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) as well as Nrf2 binding to an antioxidant response element located in the Ogg1 gene promoter. UVB exposure inhibited the phosphorylation of protein kinase B (PKB or Akt) and extracellular signal-regulated kinase (Erk), two major enzymes involved in cell protection against oxidative stress, regulating the activity of Nrf2. Akt and Erk phosphorylation was restored by phloroglucinol in the UVB-exposed keratinocytes. These results indicated that phloroglucinol attenuated UVB-induced 8-oxoG formation in keratinocytes via an Akt/Erk-dependent, Nrf2/Ogg1-mediated signaling pathway.

• Journal of Applied Biological Chemistry
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• 제65권3호
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• pp.221-230
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• 2022

본 연구는 실리마린의 항산화 및 항염증 활성을 알아보고자 하였다. 항산화 활성은 2,2-diphenyl-1-picrylhydrazyl (DPPH)와 2,2'-azino-bis (3-ethylbenzothiazoline5 6-sulfonic acid) (ABTS) 라디칼에 대한 소거능을 측정하여 확인하였다. 실리마린은 1 mg/mL의 농도에서 DPPH 라디칼을 71%, ABTS 라디칼을 78% 소거하여 우수한 항산화 효과를 나타냈다. 실리마린은 DNA의 산화적 손상을 효과적으로 억제하였고 사람 혈청 단백질과 Cu,Zn-SOD의 산화적 변형을 억제하였다. 또한 실리마린은 H₂O₂와 LPS에 의한 세포사멸, ROS 생성 및 DNA fragmentation을 억제하였다. 본 연구 결과들을 통해 실리마린은 효과적인 천연 항산화 및 항염증 소재로 적용될 수 있음을 제시하였다.

• 사상체질의학회지
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• 제19권3호
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• pp.242-256
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• 2007

1. Objectives Purpose of this study is to prove anti-aging and anti-oxidative effects of Gongjinhugwon-dan decoction. 2. Methods The SD rats used in this experiment were 6, 18, and 36 weeks old. Each age group was again divided into three groups. These nine groups consisted of 8 rats each. One group was given no treatment, another group was dosed $200{\mu}l$ of normal saline daily, and the last group was dosed $200{\mu}l$ of 1 % Gongjinhugwon-dan and saline mixture. At the conclusion of the experiment, the age groups were relabelled accordingly (10 weeks, 22 weeks, and 40 weeks). After 4 weeks, change of weight and liver markers were measured. Serum LDL cholesterol, total bilirubin, albumin, glucose, GOT and GPT levels were observed in order to check the hematological modification. Also, each organ tissue was biopsied in order to measure the SOD activity and the glutathione content change. 3. Results & Conclusions Aging did not cause any significant change in GOT and LDH, but GPT and albumin levels showed increase after GHD intake. Serum GPT was lower in the experimental group. Serum total bilirubin of the 40 w GHD group was significantly increased. The populations of dendritic cells in the spleens of the GHD groups were significantly increased. The levels of GSH in the liver of the 40 w GHD group and in the kidney of 22w-GSD were significantly increased in comparison with those of the normal groups. The degenerative change of brain tissue was decreased in the 40 w GHD group compared with those of the 40w normal group and the 40 w saline group. These results suggest that anti-oxidative GSH concentration of liver and kidney in rats treated with GHD showed significant increase in the 40 w GHD group. GHD was effective on increasing anti-oxidative substance in liver and dendritic cells in spleen, thus helping immune system and preventing cell mutation and degenerative change of brain tissues. Further studies and clinical investigation with GHD is needed.

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3213-3222
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• 2015

Background: Cancer metastasis depends on cell motility which is driven by cycles of actin polymerization and depolymerization. Reactive oxygen species (ROS) and metabolic oxidative stress have long been associated with cancer. ROS play a vital role in regulating actin dynamics that are sensitive to oxidative modification. The current work aimed at studying the effects of sub-lethal metabolic oxidative stress on actin cytoskeleton, focal adhesion and cell migration. Materials and Methods: T47D human breast cancer cells were treated with 2-deoxy-D-glucose (2DG), L-buthionine sulfoximine (BSO), or doxorubicin (DOX), individually or in combination, and changes in intracellular total glutathione and malondialdehyde (MDA) levels were measured. The expression of three major antioxidant enzymes was studied by immunoblotting, and cells were stained with fluorescent-phalloidin to evaluate changes in F-actin organization. In addition, cell adhesion and degradation ability were measured. Cell migration was studied using wound healing and transwell migration assays. Results: Our results show that treating T47D human breast cancer cells with drug combinations (2DG/BSO, 2DG/DOX, or BSO/DOX) decreased intracellular total glutathione and increased oxidized glutathione, lipid peroxidation, and cytotoxicity. In addition, the drug combinations caused a reduction in cell area and mitotic index, prophase arrest and a decreased ability to form invadopodia. The formation of F-actin aggregates was increased in treated T47D cells. Moreover, combination therapy reduced cell adhesion and the rate of cell migration. Conclusions: Our results suggest that exposure of T47D breast cancer cells to combination therapy reduces cell migration via effects on metabolic oxidative stress.

• Asian Pacific Journal of Cancer Prevention
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• 제17권9호
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• pp.4517-4525
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• 2016

Background: Sperm DNA damage is underlying aetiology of poor implantation and pregnancy rates but also affects health of offspring and may also result in denovo mutations in germ line and post fertilization. This may result in complex diseases, polygenic disorders and childhood cancers. Childhood cancer like retinoblastoma (RB) is more prevalent in developing countries and the incidence of RB has increased more than three fold in India in the last decade. Recent studies have documented increased incidence of cancers in children born to fathers who consume alcohol in excess and tobacco or who were conceived by assisted conception. The aetiology of childhood cancer and increased disease burden in these children is lin ked to oxidative stress (OS) and oxidative DNA damage( ODD) in sperm of their fathers. Though several antioxidants are in use to combat oxidative stress, the effect of majority of these formulations on DNA is not known. Yoga and meditation cause significant decline in OS and ODD and aid in regulating OS levels such that reactive oxygen speues meditated signal transduction, gene expression and several other physiological functions are not disrupted. Thus, this study aimed to analyze sperm ODD as a possible etiological factor in childhood cancer and role of simple life style interventions like yoga and meditation in significantly decreasing seminal oxidative stress and oxidative DNA damage and thereby decreasing incidence of childhood cancers. Materials and Methods: A total of 131 fathers of children with RB (non-familial sporadic heritable) and 50 controls (fathers of healthy children) were recruited at a tertiary center in India. Sperm parameters as per WHO 2010 guidelines and reactive oxygen species (ROS), DNA fragmentation index (DFI), 8-hydroxy-2'-deoxy guanosine (8-OHdG) and telomere length were estimated at day 0, and after 3 and 6 months of intervention. We also examined the compliance with yoga and meditation practice and smoking status at each follow-up. Results: The seminal mean ROS levels (p<0.05), sperm DFI (p<0.001), 8-OHdG (p<0.01) levels were significantly higher in fathers of children with RB, as compared to controls and the relative mean telomere length in the sperm was shorter. Levels of ROS were significantly reduced in tobacco users (p<0.05) as well as in alcoholics (p<0.05) after intervention. DFI reduced significantly (p<0.05) after 6 months of yoga and meditation practice in all groups. The levels of oxidative DNA damage marker 8-OHdG were reduced significantly after 3 months (p<0.05) and 6 months (p<0.05) of practice. Conclusions: Our results suggest that OS and ODD DNA may contribute to the development of childhood cancer. This may be due to accumulation of oxidized mutagenic base 8OHdG, and elevated MDA levels which results in MDA dimers which are also mutagenic, aberrant methylation pattern, altered gene expression which affect cell proliferation and survival through activation of transcription factors. Increased mt DNA mutations and aberrant repair of mt and nuclear DNA due to highly truncatred DNA repair mechanisms all contribute to sperm genome hypermutability and persistant oxidative DNA damage. Oxidative stress is also associated with genome wide hypomethylation, telomere shortening and mitochondrial dysfunction leading to genome hypermutability and instability. To the best of our knowledge, this is the first study to report decline in OS and ODD and improvement in sperm DNA integrity following adoption of meditation and yoga based life style modification.This may reduce disease burden in next generation and reduce incidence of childhood cancers.

• 대한예방한의학회지
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• 제4권2호
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• pp.205-213
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• 2000

The oxidative modification of low density lipoprotein(LDL) has been implicated in the development of atherosclerosis . Oxidized LDL(oxLDL) are found in macrophage foam cell , and it can induce an macrophage proliferation in atherosclerotic plaque. In this study, we investigated the hypothesis that Tokhwalkisaengtang(TK) may reduce atherosclerosis by lowering the oxidizability of LDL, To achieve this goal, we examined the effect of TK on LDL oxidation, nitric oxide production in murine macrophage cell line , and the effect of TK on cupuric sulfate-induced cytotoxicity. LDH release, and macrophage activity TK inhibited the generation of oxLDL from native LDL in macrophage cell culture, and decreased the release of LDH from cupric sulfate-stimulated macrophage. In other experiments, TK activated macrophase cell, and increased the survival rate, and enhanced nitric oxide production in macrophage.

• BMB Reports
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• 제28권1호
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• pp.12-16
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• 1995

The branched-chain ${\alpha}$-keto acid dehydrogenase (BCKAD) complex is a rate limiting enzyme which catalyzes the oxidative decarboxylation of branched-chain ${\alpha}$-keto acids. Numerous studies have suggested that BCKAD is subject to covalent modification in vitro via phosphorylation and dephosphorylation, which are catalyzed by a specific kinase and phosphatase, respectively. The biggest difficulty in the assay of BCKAD activity is to arrest the interconversion between the active and inactive forms. BCKAD activity was determined from fresh rat heart and liver tissues using homogenizing and assay buffers containing inhibitors of phosphatase and kinase. The results suggest that a radiochemical assay using ${\alpha}$-keto[1-$^{14}C$]-isovalerate as a substrate for the enzyme can be applied as a reliable method to determine in vitro enzyme activity with arrested interconversion between the active and inactive forms of the BCKAD complex.

• Bulletin of the Korean Chemical Society
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• 제25권5호
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• pp.711-714
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• 2004

Peroxynitrite enters erythrocytes through band 3 anion exchanger and oxidizes cytosolic proteins therein. As a protein associated with band 3, carbonic anhydrase II may suffer from peroxynitrite-induced oxidative damages. Esterase activity of carbonic anhydrase II decreased as the concentration of peroxynitrite increased. Neither hydrogen peroxide nor hypochlorite affected the enzyme activity. Inactivation of the enzyme was in parallel with the release of zinc ion, which is a component of the enzyme's active site. SDS-PAGE of peroxynitrite-treated samples showed no indication of fragmentation but non-denaturing PAGE exhibited new bands with lower positive charges. Western analysis demonstrated that nitration of tyrosine residues increased with the peroxynitrite concentration but the sites of nitration could not be determined. Instead MALDI-TOF analysis identified tryptophan-245 as a site of nitration. Such modification of tryptophan residues is responsible for the decrease in tryptophan fluorescence. These results demonstrate that peroxynitrite nitrates tyrosine and tryptophan residues of carbonic anhydrase II without causing fragmentation or dimerization. The peroxynitrite-induced inactivation of the enzyme is primarily due to the release of zinc ion in the enzyme's active site.

• 약학회지
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• 제48권2호
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• pp.130-134
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• 2004

Phellinus linteus (Hymenocaetaceae) has been used for the treatment of gastric cancer, noninsulin dependant diabetes, diarrhea, and menstrual irregularity. The antioxidative effect of cultivated fruit body of Phellinus linteus on low density lipoprotein oxidation was investigated. The MeOH and water extracts were examined by TEARS assay on human plasma LDL in vitro system. The MeOH ex. showed antioxidative activity, and then was fractionated into Precipitates, Water Insoluble fr., Water fro and ether fro The results showed that the lipophilic fractions, Water Insoluble fr. and Ether fr. of cu 1-tivated fruit body of Phellinus linteus, inhibited the oxidative modification of LDL.

• 펄프종이기술
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• 제40권3호
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• pp.9-14
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• 2008

Agar-agar extracted from red algae was used as a surface sizing material for paper after filtering, bleaching, and viscosity adjustment by oxidative depolymerization. Oxidized starches from corn and tapioca were compared to agar-based surface size in the respects of paper surface modification, strength improvement, and printability. Agar-based surface size showed high strength improvement and good printability as much as starch-based ones except film forming ability, which may be improved by adding additives, and demonstrated its great potential to be used as inexpensive and high quality surface size in future.