• Title/Summary/Keyword: Oxidative damages

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Various Fatty Acids Induce Cell Damages Differently in CYP2E1-transduced HepG2 Cells, E47 Cells, Compared to C34 Cells

  • Lee, Myoung-Sook;Bae, Myung-Ae
    • Nutritional Sciences
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    • v.9 no.1
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    • pp.9-13
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    • 2006
  • The differential effects of various fatty acids such as n-3 and n-6 types or degrees of unsaturation on the CYP2E1 induction and the production of lipid peroxidation (LPO) were investigated. The CYP2E1-transduced human hepatoma HepG2 cells (E47) were cultured in RPMI 1640 media containing different concentrations of various fatty acids up to 48 h incubation compared to 04 cells and CYP2E1-null cells. Treated fatty acids were linoleic acid (LA:n-6, C18:2), arachidonic acid (AA:n-6, C20:4) and docosahexaenoic acid (DHA:n-3, C22:6). The cell survival rate was decreased corresponding to the degree of unsaturation (LA>AA $\cong$DHA) and to LPO production in E47 and 04 cells. The four or five unsaturation degree of fatty acids, AA and DHA, caused time- and dose-dependent cell death in E47 cells but not as much as in C34 (without CYP2E1), suggesting an important role of CYP2E1 in the DHA mediated damage. In the levels of lipid peroxides (LPO), AA also elevated LPO by 3- and 5- fold compared to DHA or LA treated E47 cells. However, AA did not increase LPO until 48 h incubation in C34 cells. In conclusion, the polyunsaturated fatty acids induced CYP2E1 induction might be changed by the elevated levels of lipid peroxide (LPO) and oxidative stress through the connection of CYP2E1 and degrees of unsaturated fatty acids.

Inhibitory Effects of Marine Natural Products on Melanogenesis in B16 Melanoma Cells (B16 멜라닌 세포에서 해양소재 추출물의 멜라닌 생성 저해 효과)

  • Lee, Chan;Jang, Jung-Hee;Ahn, Eun-Mi;Park, Chan-Ik
    • The Korea Journal of Herbology
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    • v.27 no.4
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    • pp.73-80
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    • 2012
  • Objectives : Under normal condition melanin protects the skin from extracellular stimuli including ultraviolet (UV)-induced oxidative skin damages, but excess production and accumulation of melanin can induce hyperpigmentation causing esthetic problems. Therefore, in this study we tried to search for natural skin whitening materials from marine natural resources. Methods : Water and ethanol extracts of marine natural resources were prepared from Porphyra thalli (PT), Laminariae thallus (LT), Ostreae concha (OC), Sargassum thallus (ST), Undaria thallus (UT), Codium thalli (CT), Enteromorpha thalli (ET), Syngnathoides biaculeatus (SB), and Hippocampus coronatus (Hc). Their effects against UVB and ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis were investigated based on melanin formation in B16 mouse melanoma cells. The mRNA and protein expression of enzymes involved in the melanogenic process were further examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Results : Water extract of Ostreae concha (OCW/E) effectively inhibited UVB and ${\alpha}$-MSH-induced melanin production in B16 melanocytes, which seemed to be mediated by inhibition of mRNA expression of tyrosinase and tyrosinase-related protein 1 (TRP-1). In another experiment, ethanol extracts from Porphyra thalli (PTE/E), Laminariae thallus (LTE/E), Sargassum thallus (STE/E), Undaria thallus (UTE/E), Codium thalli (CTE/E), Syngnathoides biaculeatus (SBE/E), and Hippocampus coronatus (HcE/E) significantly suppressed UVB and ${\alpha}$-MSH-induced melanin formation. Furthermore, ethylacetate fraction isolated form LTE/E (LTE/EEt) decreased UVB and ${\alpha}$-MSH-elevated extracellular melanin levels via inhibition of tyrosinase protein expression. Conclutions : These results suggest that marine natural resources such as Porphyra thalli, Laminariae thallus, Ostreae concha, Sargassum thallus, Undaria thallus, Codium thalli, Syngnathoides biaculeatus and Hippocampus coronatus have anti-melanogenic effects, thereby exhibiting high potentials to be utilized as one of the ingredients for the development of new whitening functional cosmetics.

Molecular Analyses of the Metallothionein Gene Family in Rice (Oryza sativa L.)

  • Zhou, Gongke;Xu, Yufeng;Li, Ji;Yang, Lingyan;Liu, Jin-Yuan
    • BMB Reports
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    • v.39 no.5
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    • pp.595-606
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    • 2006
  • Metallothioneins are a group of low molecular mass and cysteine-rich metal-binding proteins, ubiquitously found in most living organisms. They play an important role in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting against intracellular oxidative damages. Analysis of complete rice genome sequences revealed eleven genes encoding putative metallothionein (OsMT), indicating that OsMTs constitute a small gene family in rice. Expression profiling revealed that each member of the OsMT gene family differs not only in sequence but also in their tissue expression patterns, suggesting that these isoforms may have different functions they perform in specific tissues. On the basis of OsMT structural and phylogenetic analysis, the OsMT family was classified as two classes and class I was subdivided into four types. Additionally, in this paper we also present a complete overview of this family, describing the gene structure, genome localization, upstream regulatory element, and exon/intron organization of each member in order to provide valuable insight into this OsMT gene family.

Protective Effects of Samul-tang on ${H_2O_2}-induced$ Cell Apoptosis in Cultured Cardiomyoblast Cells ($H_2O_2$에 의한 배양심근세포고사에 미치는 사물탕의 방어효과)

  • 박종운;한상혁;김도환;문병순
    • The Journal of Korean Medicine
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    • v.22 no.4
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    • pp.58-68
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    • 2001
  • Objectives : This study was designed to investigate the protective mechanisms of Samul-tang (SMT) on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Methods : The cultured cells were pretreated with SMT and exposed to $H_2O_2$. The cell damage was assessed by using MTT assay. Also, we used Hoechst staining, Western blotting analysis. Results : SMT significantly reduced both $H_2O_2$-induced cell death and chromatin fragmentation. The decrease of Bcl2 expression by $H_2O_2$ was inhibited by SMT. In addition, the increase of Bax expression was also inhibited by SMT. In particular, Fas expression, which is generally recognized as cell death inducing signal by Fas/FasL interaction, was markedly decreased by $H_2O_2$ in a time-dependent manner, whereas this decrease was completely prevented by SMT. The cotreatment of SMT and $H_2O_2$ in H9c2 cells also induced the phosphorylation of ERK in a time-dependent manner. Moreover, PD098059, a specific inhibitor of ERKl/2, attenuated the protective effect of SMT on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Furthermore, the protective effect of SMT was significantly blocked by treatment of SB203580, a specific inhibitor of p38. Conclusions : Taken together, this study suggests that the protective effects of the water extract of SMT against oxidative damages may be mediated by the modulation of Bel2 and Bax expression via the regulation of ERK and p38 signaling pathway.

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A Study of Neuroproctective Effect of Bupleuri Radix on Hippocampal Neurons (시호(柴胡)의 뇌해마 신경세포 보호효능에 대한 연구)

  • Lee, Won-Chul;Shin, Kwang-Sik
    • The Journal of Internal Korean Medicine
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    • v.25 no.4
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    • pp.227-241
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    • 2004
  • Objective : This study was performed to investigate neuroprotective effects of Bupleuri Radix against oxidative and ischemic damages. Method : To observe the neuroprotective effects against ischemic damage, ischemic insult was induced by oxygen/glucose deprivation (OGD) on organotypic hippocampal slice cultures (OHSC) from 1 week-old Sprague-Dawley rats. Propidium iodide (PI) fluorescence-stained neuronal dead-cell areas, area percentages and TUNEL-positive apoptotic cells in CA1 and dentate gyrus, and LDH levels in culture media of the OHSC were measured following Bupleuri Radix extract treatment. Result : The following results were obtained: (1) The $5\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in CA1 region of the OHSC from 18 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ of Bupleuri Radix treatment was also significant from 6 hrs to 48 hrs following the OGD and was more effective. (2) The 5 and $50\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in DG region of the OHSC from 6 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ treatment was more effective than the $5\;{\mu}g/ml$ treatment. (3) Bupleuri Radix treatment demonstrated a significant decrease in TUNEL-positive apoptotic cells in CA1 region (with 5 and $50\;{\mu}g/ml$) and in DG region (with $50\;{\mu}g/ml$) of the OHSC damaged by the OGD. (4) Bupleuri Radix treatment demonstrated a significant decrease in LDH concentrations in culture media of the OHSC damaged by the OGD. Conclusion : These results suggest that Bupleuri Radix has neuroprotective and control effects on inflammatory and immune responses where there has been ischemic damage to the central nervous system.

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Effects of Mangeum-tang and Gamimangeum-tang on the Cultured Spinal Dorsal Root Ganglion Cells (만금탕 및 가미만금탕이 배양 척수후근신경절 세포에 미치는 영향)

  • Choi Gyu Seon;Yun Sang Hak;Yeom Seung Ryong;Lee Su kyung;Shin Byung Cheul;Kwon Young Dal;Song Yung Sun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.503-509
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    • 2003
  • The purpose of this study is to examine the toxic effects caused by xanthine oxidase/hypoxanthine (XO/HX) and the effects of herbal extracts such as Mangeum-tang (萬金湯: MGT) and Gamimangeum-tang (加味萬金湯: GMGT) on the treatment of the toxic effects. The results of these experiments were XO/HX, an oxygen radical-generating system, decreased the survival rates of the cultured cells on XTT assay, the amount of DNA syntheses, and the amount of neurofilaments, and increased c-fos positive cells, MGT and GMGT have the efficacy of increasing the survival rates of the cultured cells by increasing the amount of neurofilaments and DNA synthesis and decreasing the c-fos positive cells damaged by XO/HX, From the above results, it is suggested that MGT and GMGT have marked efficacy as a treatment for the damages caused by the XO/HX-mediated oxidative stress. And MGT and GMGT are thought to have certain pharmacological effects.

Antioxidant and Cytotoxicity Effects of Luteolin

  • Park Jin-Ny;Kang Kyoung-Ah;Zhang Rui;Piao Mei Jing;Park So-Yoon;Kim Ju-Sun;Kang Sam-Sik;Hyun Jin-Won
    • Toxicological Research
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    • v.22 no.4
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    • pp.391-395
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    • 2006
  • Flavonoids are polyphenolic compounds that are ubiquitous in plants. They have been shown to possess a variety of biological activities, such as antioxidant and anticancer. Reactive oxygen species (ROS) lead to damages of cellular molecules and it is the one of various factors to induce cancer. The one of flavonoids, Luteolin, was found to scavenge 1,1-diphenyl-2-piculhydrazyl (DPPH) radical and intracellular reactive oxygen species (ROS). Moreover luteolin showed protection on hamster lung fibroblast cells (V79-4 cell) damage induced by $H_2O_2$. And then it was investigated whether it may show cytotoxicity effect against various cancer cells by MTT, Luteolin at $10{\mu}g/ml$ showed the cell viability of 63.2%, 34.7%, 18.4% and 71.4% against NCl-H460, HeLa, U937 and MCF-7, respectively. As a result, luteolin shows more sensitive to U937 cells among the tested cancer cell lines. In summary, luteolin has antioxidant and cytotoxicity effect or various cancer cell lines.

Effects of red ginseng total saponin on Menadione-induced hepatotoxicity in the rat (Menadione에 의해 유발된 간독성에 미치는 홍삼사포닌의 영향)

  • Jang, Bong-jun;Bae, Chun-sik;Cho, Yong-seong;Cha, Yong-ho;Park, Chang-won;Cho, Tae-hyun;Chang, Kyung-jin
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.619-627
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    • 1997
  • It is known that 2-methyl-1,4-naphtoquinone(menadione, MD) induces hepatotoxicities both in vivo and in vitro. These toxic effects are believed to result from oxidative damages to hepatocytes by "active oxygen" species via one-electron reduction of the naphtoquinone. The ginsenoside(GS) is a complex mixture of individual ginsenosides which is known to produce a range of effects on the cardiovascular and central nervous systems. In particular, GS has an antioxidant effect. In this experiment we studied the effect of GS from red panax ginseng(red ginseng total saponin, RGTS) on free radical-induced liver injuries by MD. Administration of MD($150{\mu}M$) caused an increase in aspartate aminotransferase(AST) activities and lipid peroxidation, decrease in alkaline phosphatase(ALP) activities and total bilirubin levels in blood, caused depletion of GSH and changes of antioxidant enzyme(superoxide dismutase, catalase) activities are shown in liver tissue. Administration of RGTS restored the AST levels that increased by MD, but catalase showed no significant changes. RGTS also had an effect of restoring the GSH level and had some synergistic effects with SOD. These data suggest that RGTS may have some protective effects on liver injury which is related with the oxygen free radical.

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Formation of DNA-Protein Crosslink at Oxidized Abasic Site Mediated by Human DNA Polymerase Iota and Mitochondrial DNA Polymerase Gamma

  • Son, Mi-Young;Jun, Hyun-Ik;Goo, Sun-Young;Sung, Jung-Suk
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.1-8
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    • 2009
  • Human genomic DNA is continuously attacked by oxygen radicals originated from cellular metabolic processes and numerous environmental carcinogens. 2-deoxyribonolactone (dL) is a major type of oxidized abasic (AP) lesion implicated in DNA strand scission, mutagenesis, and formation of covalent DNA-protein crosslink (DPC) with DNA polymerase (Pol) ${\beta}$. We show here that human DNA polymerase (Pol)${\iota}$ and mitochondrial $Pol{\gamma}$ give rise to stable DNA-protein crosslink (DPC) formation that is specifically mediated by dL lesion. $Pol{\gamma}$ mediates DPC formation at the incised dL residue by its 5'-deoxyribose-5-phosphate (dRP) lyase activity, while $Pol{\gamma}$ cross links with dL thorough its intrinsic dRP lyase and AP lyase activities. Reactivity in forming dL-mediated DPC was significantly higher with $Pol{\gamma}$ than with $Pol{\iota}$. DPC formation by $Pol{\gamma}$, however, can be reduced by an accessory factor of $Pol{\gamma}$ holoenzyme that may attenuate deleterious effects of crosslink adducts on mitochondrial DNA. Comparative kinetic analysis of DPC formation showed that the rate of DPC formation with either $Pol{\iota}$ or $Pol{\gamma}$ was lower than that with $Pol{\beta}$. These results revealed that the activity of catalytic lyase in DNA polymerases determine the efficiency of DPC formation with dL damages. Irreversible crosslink formation of such DNA polymerases by dL lesions may result in a prolonged strand scission and a suicide of DNA repair proteins, both of which could pose a threat to the genetic and structural integrity of DNA.

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Neuroprotective and Anti-inflammatory Effects of Phenolic Compounds in Panax ginseng C.A. Meyer (인삼에 함유된 페놀성 선분의 신경세포보호 및 항염증 효과)

  • Kong, Yeon-Hee;Lee, Young-Chul;Choi, Sang-Yoon
    • Journal of Ginseng Research
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    • v.33 no.2
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    • pp.111-114
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    • 2009
  • The six phenolic-compound (ascorbic acid, maltol, esculetin,p-coumaric acid, cinnamic acid, and quercetin) contents of Panax ginseng c.A. Meyer were determined in this study. The results showed that the ascorbic acid, cinnamic acid, and esculetin contents of Panax ginseng C.A. Meyer are higher than those of the other ingredients. Among these compounds, ascorbic acid and cinnamic acid significantly inhibited LPS-induced nitric oxide production in the RAW 264.7 cells. Cinnamic acid also effectively inhibited the oxidative damages in the human neuroblastoma SH-SY5Y cells. Although this study examined the neuroprotective and anti-inflammatory activities using only one kind of cells, its results suggest that cinnarnic acid potently contributes to the neuroprotective and anti-inflammatory properties of Panax ginseng C.A. Meyer.