• 제목/요약/키워드: Antioxidative enzyme

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Curcumin-Induced Apoptosis of A-431 Cells Involves Caspase-3 Activation

  • Shim, Joong-Sup;Lee, Hyung-Joo;Park, Sang-shin;Cha, Bong-Gee;Chang, Hae-Ryong
    • BMB Reports
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    • v.34 no.3
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    • pp.189-193
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    • 2001
  • Curcumin a yellow pigment from Curcuma Tonga, has been known to possess antioxidative and anticarcinogenic properties, as well as to induce apoptosis in some cancer cells. There have been, however, several contradictory reports that hypothesized curcumin (a hydrophobic molecule) can bind a membrane Gpid bilayer and induce nonspecific cytotoxicity in some cell lines. Why curcumin shows these contradictory effects is unknown. In A-431 cells, growth inhibition by curcumin is due mostly to the specific inhibition of the intrinsic tyrosine kinase activity of the epidermal growth factor receptor, as reported earlier by Korutla et al. Thus, we assumed that the cell death of A-431 by curcumin might be due to the specific induction of apoptosis. In this paper we clearly show that curcumin induces apoptosis in A-431 cells. The cureumin-induced cell death of A-431 exhibited various apoptotic features, including DNA fragmentation and nuclear condensation. Furthermore, the curcumin-induced apoptosis of A-431 cells involved activation of caspase-3-like cysteine protease. Involvement of caspase-3 was further confirmed by using a caspase-3 specific inhibitor, DEVD-CHO. In another study, decreased nitric oxide (NO) production was also shown in A-431 cells treated with curcumin, which seems to be the result of the inhibition of the iNOS expression by curcumin, as in other cell lines. However, 24 h after treatment of curcumin there was increased NO production in A-431 cells. This observation has not yet been clearly explained. We assumed that the increased NO production may be related to denitrosylation of the enzyme catalytic site in caspase-3 when activated. Taken together, this study shows that the cell death of A-431 by curcumin is due to the induction of apoptosis, which involves caspase-3 activation.

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Ethanol Eluted Extract of Rhus verniciflua Stokes Showed both Antioxidant and Cytotoxic Effects on Mouse Thymocytes Depending on the Dose and Time of the Treatment

  • Lee, Jeong-Chae;Kim, Ju;Lim, Kye-Taek;Yang, Moon-Sik;Jang, Yong-Suk
    • BMB Reports
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    • v.34 no.3
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    • pp.250-258
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    • 2001
  • For a long time Rhus verniciflua Stokes (RVS) has traditionally been used as a herbal plant. It is known to contain various biological activities. Previously, a crude ethanol extract from RVS was reported to have antioxidant effects, and antiproliferative activities, on human cancer cell lines. In this report, we prepared a highly purified ethanol extract from RVS, which did not contain the urushiol derivatives, named REEE-1 ($\underline{R}$hus $\underline{e}$thanol $\underline{e}$luted $\underline{e}$xtract-1), to investigate the mechanisms of the scavenging activity of hydroxyl radicals using mouse thymocytes. The results from the deoxyribose, DNA nicking, and glucose/glucose oxidase enzyme assays showed that REEE-1 contained a strong scavenging activity of oxygen free radicals, especially of hydroxyl radicals. However, interestingly, REEE-1 also showed cytotoxicity against the thymocytes, although the effect was variable, depending on the concentrations and times of treatment. The REEE-1-mediated cytotoxicity against thymocytes, which has been used as one of the well-characterized models for apoptosis studies, was verified to be apoptotic. This was proven by the following: the appearance of DNA laddering, increases in DNA fragmentation, low fluorescence intensity in the nuclei after propidium iodide staining, and positive Annexin V staining of the cells. These results suggested that REEE-1 had both antioxidative activity and cytotoxicity against the thymocytes, although the effect of the cytotoxicity was variable, depending on the dose and time of the treatment.

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The effect of carrot juice, ${\beta}$-carotene supplementation on lymphocyte DNA damage, erythrocyte antioxidant enzymes and plasma lipid profiles in Korean smoker

  • Lee, Hye-Jin;Park, Yoo-Kyoung;Kang, Myung-Hee
    • Nutrition Research and Practice
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    • v.5 no.6
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    • pp.540-547
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    • 2011
  • High consumption of fruits and vegetables has been suggested to provide some protection to smokers who are exposed to an increased risk of numerous cancers and other degenerative diseases. Carrot is the most important source of dietary ${\beta}$-carotene. Therefore, the objective of this study was to investigate whether carrot juice supplementation to smokers can protect against lymphocyte DNA damage and to compare the effect of supplementationof capsules containing purified ${\beta}$-carotene or a placebo (simple lactose). The study was conducted in a randomized and placebo-controlled design. After a depletion period of 14 days, 48 smokers were supplemented with either carrot juice (n = 18), purified ${\beta}$-carotene (n = 16) or placebo (n = 14). Each group was supplemented for 8 weeks with approximately 20.49 mg of ${\beta}$-carotene/day and 1.2 mg of vitamin C/day, as carrot juice (300 ml/day) or purified ${\beta}$-carotene (20.49 mg of ${\beta}$-carotene, 1 capsule/day). Lymphocyte DNA damage was determined using the COMET assay under alkaline conditions and damage was quantified by measuring tail moment (TM), tail length (TL), and% DNA in the tail. Lymphocyte DNA damage was significantly decreased in the carrot juice group in all three measurements. The group that received purified ${\beta}$-carotene also showed a significant decrease in lymphocyte DNA damage in all three measurements. However, no significant changes in DNA damage was observed for the placebo group except TM (P = 0.016). Erythrocyte antioxidant enzyme was not significantly changed after supplementation. Similarly plasma lipid profiles were not different after carrot juice, ${\beta}$-carotene and placebo supplementation. These results suggest that while the placebo group failed to show any protective effect, carrot juice containing beta-carotene or purified ${\beta}$-carotene itself had great antioxidative potential in preventing damage to lymphocyte DNA in smokers.

The Effects of Pulsed Electromagnetic Fields on Blood components, Antioxidant enzymes and Reactive Oxygen in Hyperlipidemic Rats (맥동전자장이 고지혈증 흰쥐의 혈액 성분, 항산화 효소 및 활성 산소에 미치는 영향)

  • Bang, Hyun-Soo;Jeong, In-Ho;Lee, Sang-Deok
    • Journal of Digital Convergence
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    • v.12 no.7
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    • pp.349-356
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    • 2014
  • The present study was aimed to investigate the effects of the application of pulsed electromagnetic fields on the blood components associated hyperlipidemia, antioxidant enzymes and reactive oxygen. The subjects were divided into three groups: General Diet, High fat Diet, High fat diet and pulsed electromagnetic fields. Pulsed electromagnetic field was applied with pulsed electromagnetic energy therapy equipment. The Glucose, free fatty acids, triglycerides, cholesterol and insulin was used to measure hyperlipidemia-related blood components, and the GSH, GRD, XO and MDA was used to measure antioxidant enzymes and reactive oxygen. The applied pulsed electromagnetic field has improved the concentrations of blood components and increasing GSH and GRD, that were decreased due to a high-fat diet and by reducing the increased MDA and XO to a level of general diet group. Therefore, the applied pulsed electromagnetic field can be the effective treatment for changes in the blood composition of hyperlipidemic rats, enhancement of antioxidative activity and the inhibition of reactive oxygen.

Effects of Hexaconazole on Growth and Antioxidant Potential of Cucumber Seedlings under UV-B Radiation

  • Kim, Tae-Yun;Hong, Jung-Hee
    • Journal of Environmental Science International
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    • v.21 no.12
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    • pp.1435-1447
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    • 2012
  • The present study was conducted to determine the effect of hexaconazole (HEX), a triazole fungicide, on the growth, yield, photosynthetic response and antioxidant potential in cucumber (Cucumis sativus L.) plants subjected to UV-B stress. UV-B radiation and HEX were applied separately or in combination to cucumber seedlings. The growth parameters were significantly reduced under UV-B treatment, however, this growth inhibition was less in HEX treated plants. HEX caused noticeable changes in plant morphology such as reduced shoot length and leaf area, and increased leaf thickness. HEX was quite persistent in inhibiting shoot growth by causing a reduction in shoot fresh and dry weight. HEX noticeably recovered the UV-B induced inhibition of biomass production. Significant accumutation in anthocyanin and flavonoid pigments in the leaves occurred as a result of HEX or UV-B treatments. HEX permitted the survival of more green leaf tissue preventing chlorophyll content reduction and higher quantum yield for photosystemII under UV-B exposure. HEX treatment induced a transient rise in ABA levels in the leaves, and combined application of HEX and UV-B showed a significant enhancement of ABA content which activates $H_2O_2$ generation. UV-B exposure induced accumulation of $H_2O_2$ in the leaves, while HEX prevented UV-B induced increase in $H_2O_2$, indicating that HEX serves as an antioxidant agent able to scavenge $H_2O$ to protect cells from oxidative damage. An increase in the ascorbic acid was observed in the HEX treated cucumber leaves affecting many enzyme activities by removing $H_2O_2$ during photosynthetic processes. The activities of antioxidant enzymes including catalase(CAT), ascorbate peroxidase(APX), superoxide dismutase(SOD) and peroxidase(POD) in the leaves in the presence of HEX under UV-B stress were higher than those under UV-B stress alone. These findings suggest that HEX may participate in the enhanced tolerance to oxidative stress. From these results it can be concluded that HEX moderately ameliolate the effect of UV-B stress in cucumber by improving the components of antioxidant defense system.

Oxidative Stress and Antioxidant Activities of Intertidal Macroalgae in Korea

  • Park, Jung-Jin;Han, Tae-Jun;Choi, Eun-Mi
    • Preventive Nutrition and Food Science
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    • v.16 no.4
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    • pp.313-320
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    • 2011
  • The oxidative stress level and antioxidant activities in two green algae (Ulva pertusa and Ulva linza), two brown algae (Agarum cribrosum and Dictyota dichotoma), and three red algae (Grateloupia lanceolata, Carpopeltis affinis, and Gracilaria verrucosa) collected from intertidal regions of Korea were assessed. In the two green algae, although the total glutathione content was not as high as that of the brown algae, the glutathione pool was extremely reduced, and the glutathione reductase (GRd)/glutathione peroxidase (GPx) activity ratio was high, which apparently plays an important role for protection against oxidative damage, as manifested by low lipid peroxidation. In the brown algae, which exhibited a low lipid peroxidation level that was comparable to the green algal species, the highest glutathione content, together with high GPx activity, appears to be the most important factor in their antioxidant protection. The red algal species exhibited extremely high lipid peroxidation levels. They also contained the lowest and most oxidized glutathione among the species, as well as the lowest GRd activity. In spite of the marked difference in the glutathione content, the significant difference in the activity of ${\gamma}$-glutamylcysteine ligase, the rate limiting enzyme for glutathione synthesis, among the species was not exhibited. Our results suggest that there is a significant difference in the levels of oxidative stress and antioxidant capacity among the algal species, and that the glutathione system, especially the efficiency of glutathione recycling, plays a vital role in antioxidative protection in algal species.

Bioactivities of the Herb Extracts Used for Gamhongroju, a Korean Liqueur

  • Lee, Sae-Rom;Jung, Ha-Na;Cho, Hyunn-Ho;Jhin, Chang-Ho;Hwang, Keum-Taek;Jeong, Seok-Tae;Kim, Tae-Young
    • Preventive Nutrition and Food Science
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    • v.16 no.4
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    • pp.333-338
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    • 2011
  • In this study, antioxidative and anti-inflammatory activities of the herb (cinnamon, clove, glehnia root, ginger, violet-root cromwell, licorice, citrus peel and longan) extracts used for gamhongroju, one of the popular liqueurs in Korea, were investigated. Twenty grams of individual herbs were extracted in 60% purified ethanol and freeze-dried. A mixture of the individual herb extracts (HEM) was separately prepared. Cytotoxicity of the individual extracts and HEM on murine RAW264.7 macrophage cells were examined along with their recovering activity on $H_2O_2$-treated RAW264.7 cells. Antioxidant and anti-inflammatory activities of the extract-treated cells were determined by measuring superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, and Trolox equivalent antioxidant capacity (TEAC), nitric oxide (NO) and prostaglandin E2 (PGE2) levels. Violet-root cromwell extract showed the least cytotoxicity in terms of treated concentration. Most of the extracts, below levels of cytotoxicity, recovered the $H_2O_2$-treated cells. Treatment with some of the extracts increased SOD and GPx activities and TEAC levels while a majority inhibited the production of NO and PGE2 in lipopolysaccharide (LPS)-treated cells.

Effect of Gamisogunjung-tang on Antioxidation Activity in Rats Induced Aging by D-Galactose (가미소건중탕(加味小建中湯)이 D-galactose로 노화(老化)를 유발(誘發)시킨 백서(白鼠)의 항산화능(抗酸化能)에 미치는 영향(影響))

  • Park, Sun-Young;Back, Jung-Han;Seo, Jung-Min
    • The Journal of Pediatrics of Korean Medicine
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    • v.19 no.1
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    • pp.153-171
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    • 2005
  • Objective : This experiment has been done to evaluate the effects of Gamisogunjung-tang(GST) on antioxidant capability and lipidic concentration in blood which are presumed to be related to aging. Method : In this study, we divided 14 weeks old SD rats into normal group, control group and GST group. Control and GST group were induced aging by D-galactose. At the same time GST group were administered extract of Gamisogunjung-tang for 6 weeks. After then we took blood, and measured the activities of SOD, GSH-px, catalase in erythrocytes and measured TBARS values, concentrations of total lipid, tryglyceride, total cholesterol, HDL-cholesterol in plasma. Results : The activities of SOD, GSH-px, catalase in erythrocytes increased significantly in GST group compared with control group. The value of TBARS and the concentration of total lipid, total cholesterol in plasma decreased significantly in GST group compared with control group. The concentration of HDL-cholesterol increased significantly in GST group compared with control group. The concentration of triglyceride were not noticeable. Conclusion : it is considered that Gamisogunjung-tang has an influence on control aging by activation the antioxidative enzyme systems in erythrocytes and decreasing the concentration of lipid in blood plasma.

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The Preventive Inhibition of Chondroitin Sulfate Against the $CCl_4$-Induced Oxidative Stress of Subcellular Level

  • Lee, Jin-Young;Lee, Sang-Hun;Kim, Hee-Jin;Ha, Jong-Myung;Lee, Sang-Hyun;Lee, Jae-Hwa;Ha, Bae-Jin
    • Archives of Pharmacal Research
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    • v.27 no.3
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    • pp.340-345
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    • 2004
  • Our work in this study was made in the microsomal fraction to evaluate the lipid peroxidation by measuring superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and malondialdehyde (MDA) and to elucidate the preventive role of CS in the $CCl_4$-induced oxidative stress. The excessive lipid peroxidation by free radicals derived from $CCl_4$ leads to the condition of oxidative stress which results in the accumulation of MDA. MDA is one of the end-products in the lipid peroxidation process and oxidative stress. MDA, lipid peroxide, produced in this oxidative stress causes various diseases related to aging and hepatotoxicity, etc. Normal cells have a number of enzymatic and nonenzymatic endogenous defense systems to protect themselves from reactive species. The enzymes in the defense systems, for example, are SOD, CAT, and GPx. They quickly eliminate reactive oxygen species (ROS) such as superoxide anion free radicalㆍO$^{[-10]}$ $_2$, hydrogen peroxide $H_2O$$_2$ and hydroxyl free radicalㆍOH. CS inhibited the accumulation of MDA and the deactivation of SOD, CAT and GPx in the dose-dependent and preventive manner. Our study suggests that CS might be a potential scavenger of free radicals in the oxidative stress originated from the lipid peroxidation of the liver cells of $CCl_4$-treated rats.

Protective Effects of the BuOH Fraction from Laminaria japonica Extract on High Glucose-induced Oxidative Stress in Human Umbilical Vein Endothelial Cells

  • Park, Min-Jung;Song, Young-Sun;Han, Ji-Sook
    • Preventive Nutrition and Food Science
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    • v.11 no.2
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    • pp.94-99
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    • 2006
  • This study investigated the protective effect of the butanol (BuOH) fraction from Laminaria japonica (BFLJ) extract on high glucose-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). Freeze-dried L japonica was extracted with distilled water, and the extracted solution was mixed with ethanol then centrifuged. The supernatant was subjected to sequential fractionation with various solvents. The BuOH fraction was used in this study because it possessed the strongest antioxidant activity among the various solvent fractions. To determine the protective effect of the BFLJ, oxidative stress was induced by exposing of HUVECs to the high glucose (30 mM) or normal glucose (5.5 mM) for 48 hr. Cell viability, lipid peroxidation, glutathione (GSH) concentration, and antioxidant enzyme activities such as catalase, superoxide dismutase (SOD), glutathione peroxidase (GSH-px), and glutathion reductase (GSH-re) were measured. Exposure of HUVECs to high glucose for 48 hr resulted in a significant (p<0.05) decrease in cell viability, SOD, GSH-px and GSH-re and a significant (p<0.05) increase in thiobarbituric acid reactive substances (TBARS) formation in comparison to the cells treated with 5.5 mM glucose or untreated with glucose. BFLJ treatment decreased TBARS formation and increased cell viability, GSH concentration, and activities of antioxidant enzymes including catalase, SOD, GSH-px, and GSH-re in high glucose pretreated HUVECs. These results suggest that BFLJ may be able to protect HUVECs from high glucose-induced oxidative stress, partially through the antioxidative defence systems.