• Title/Summary/Keyword: ROS-scavenging enzymes

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Overexpression of OsNAC17 enhances drought tolerance in rice

  • Kim, Tae Hwan;Kim, Ju-Kon
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.168-168
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    • 2017
  • Drought conditions during cultivation reduce agricultural production yield less than a theoretical maximum yield under normal condition. Plant specific NAC transcription factors in rice are known to play an essential roles in stress resistance transcriptional regulation. In this study, we report the rice (Oryza sativa L japonica) NAM, AFTF and CUC transcription factor OsNAC17, which is predominantly induced by abiotic stress in leaf, was contribute to the drought tolerance mediated reactive oxygen species (ROS) in transgenic rice plants. Constitutive (PGD1) promoter was introduced to overexpress OsNAC17 and produced the transgenic PDG1:OsNAC17. Overexpression of OsNAC17 throughout the whole plant improved drought resistance phenotype at the vegetative stage. Morphological characteristics such as grain yield, grain filling rate, and total grain weight improved by 22~64% over wild type plants under drought conditions during the reproductive stage. The improved drought tolerance in transgenic rice was involved in reducing stomatal density up to 15% than in wild type plants and in increasing reactive oxygen species-scavenging enzyme. DEG profiling experiment identified 119 up-regulated genes by more than twofold (P<0.01). These genes included UDP-glycosyltransferase family protein, similar to 2-alkenal reductase (NADPH-dependent oxireductase), similar to retinol dehydrogenase 12, Lipoxygenase, and NB-ARC domain containing protein related in cell death. Furthermore, OsNAC17 was act as a transcriptional activator, which has an activation domain in C-terminal region. These result demonstrate that the overexpression of OsNAC17 improve drought tolerance by regulating ROS scavenging enzymes and by reducing stomatal density

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Photochemical Damage and Responses of Antioxidant Enzymes in Rice Leaves Induced to Light-Chilling (Light-chilling에 의해 유도된 벼 잎에서의 광합성 변화와 항산화 효소의 반응)

  • Koo, Jeung-Suk;Choo, Yeon-Sik;Lee, Chin-Bum
    • Journal of Life Science
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    • v.19 no.4
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    • pp.442-448
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    • 2009
  • We investigated photooxidation and responses of antioxidant enzymes involved in scavenging reactive oxygen species (ROS) after light-chilling ($4^{\circ}C$) for 2 days and post chilling ($25^{\circ}C$) in rice leaves. Chilling leaves indicated a 50% reduction in photosynthetic efficiency ($F_v/F_m$ ratio) and a 48% increase of $H_2O_2$, respectively, compared to the control group. In comparison with the control, activities of superoxide dismutase (SOD) and glutathione reductase (GR) increased at light-chilling and post-chilling. CuZn-SOD and Mn-SOD among SOD forms were detected in rice leaves, while Fe-SOD was not found. The increase of SOD and GR activity may serve as a basis for defense against chilling injury as it dismutase superoxide generated by light-chilling. Catalase (CAT) activity decreased during light-chilling, while activity of APX showed remarkable increase during light-chilling in rice leaves. Among CAT isoforms analyzed by 10% native PAGE, activities of isoform -2 and -3 were inhibited during light-chilling. From the elevated APX activity and decreased CAT activity, we suggest that these two enzymes show mutual supplementary relationships, indicating different tendency during light-chilling.

Inhibitory Effects of Allium sacculiferum Max. Methanol Extracts on ROS Production and Lipid Accumulation during Differentiation of 3T3-L1 Cells (참산부추(Allium sacculiferum Max.) 메탄올 추출물의 지방세포 내 ROS 생성 및 지질 축적 억제 효능)

  • Choi, Hye-Young;Kim, Gun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.6
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    • pp.822-828
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    • 2014
  • Allium sacculiferum Max. (ASM) is a perennial plant of the Liliaceae family and grows over the entire regions of Korea. Obesity is a serious health problem worldwide and has currently become a prevalent chronic disease. Adipocytes produced by preadipocyte differentiation during adipogenesis and adipocytes combined with abnormal accumulation cause obesity. Recently, intracellular reactive oxygen species (ROS) were shown to accelerate lipid accumulation in 3T3-L1 cells. In this study, we investigated the effects of ASM methanol extracts on ROS production and lipid accumulation in 3T3-L1 adipocytes. Our results indicate that the 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of ASM methanol extracts increased in a dose-dependent manner. ASM methanol extracts suppressed ROS production and lipid accumulation during adipogenesis. In addition, ASM methanol extracts inhibited the mRNA expression of both pro-oxidant enzymes such as glucose-6-phosphate dehydrogenase as well as the transcription factors, including sterol regulatory element-binding proteins 1c, peroxisome proliferator-activated receptor ${\gamma}$, and CCAAT/enhancer-binding protein ${\alpha}$. Our results suggest that ASM methanol extracts inhibit ROS production and lipid accumulation by controlling ROS regulatory genes and adipogenic transcription factors. Thus, ASM has potent natural antioxidant, anti-adipogenic properties and have potential in the development of a potent anti-obesity agent.

Protection by Sunghyangchungisan against Oxidative Endothelial Cell Injury (배양(培養)된 혈관(血管) 내피세포(內皮細胞)에서 산화성(酸化性) 세포(細胞) 손상(損傷)에 미치는 성향정기산(星香正氣散)의 보호(保護) 효과(效果))

  • Lee Dong-Uhn;Kim Young-Kyun
    • Herbal Formula Science
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    • v.8 no.1
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    • pp.147-167
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    • 2000
  • Reactive oxygen species (ROS) play an important role in the pathogenesis of a variety of life threatening conditions such as atherosclerosis, myocardial infarction and cerebral stroke. In this study, the effect of Sunghyangchungisan (SHCS) as a cytoproctant against ROS-induced cell injury was studied by investigating its effect on $H_{2}O_2-induced$ cell injury in cultured endothelial cells derived from the human umbilical vein. SHCS effectively proteced the cells against $H_{2}O_2-induced$ injury determined by trypan blue exclusion ability and lactate dehydrogenase (LDH) release. The effect of SHCS was concentration-dependent and the concentrations to inhibit by 50% the cell death and LDH release were $0.9{\pm}0.1$ and $1.2{\pm}0.1\;mg/ml$, respectively. In addition, SHCS effectively protected the cells against t-butylhydroperoside- and menadione-Induced injury as well. SHCS inhibited lipid peroxidation determined by malondialdehyde production. SHCS exerted as an effective scavenger of ROS produced by exposing the cells to $H_{2}O_2$ The activities of the intracellular ROS scavenging enzymes such as superoxide dismutase, catalase and glutathione peroxidase were not Influenced by SHCS.These results indicate that SHCS might exert as an effective cytoprotectant against ROS-induced cell injury. Further intensive studies would provide us insights into mechanisms of the pharmacological actions of SHCS.

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Peroxynitrite Scavenging Mechanism of Ojawhan (오자환(五子丸)의 Peroxynitrite 제거 작용)

  • Kim, Hyung-Joon;Jeong, Ji-Cheon
    • The Journal of Internal Korean Medicine
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    • v.26 no.1
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    • pp.107-118
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    • 2005
  • Objectives : Peroxynitrite $(ONOO^-)$, fonned from the reaction of $O_2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. Due to the lack of endogenous enzymes to thwart $ONOO^-$ activation, developing a specific $ONOO^-$ scavenger is remarkably important. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_2^-$ and its scavenging mechanism of Ojawhan. Methods : To investigate scavenging activities of $ONOO^-$, NO, $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on Ojawhan was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. Oxidative stress was induced by strong oxidants t-butyl hydroperoxide (t-BHP). Endothelial cell (YPEN-1) was used for detection of intracellular oxidative stress. Results : Ojawhan markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthennore, ${\underline{Ojawhan}}$ blocked lipopolysaccharide (LPS)-induced $ONOO^-$, $O_2^-$ and NO generation utilizing kidney homogenates of LPS-injected mouse and inhibited t-BHP-induced ROS and $ONOO^-$ in endothelial cell culture system. Conclusions : These results suggest that Ojawhan be developed as an effective $ONOO^-$ scavenger for the prevention of $ONOO^-$ involved diseases and age-related diseases.

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Effects of Chicken Treated with Hwangki-Beni Koji Sauces on ROS Generating and Scavenging Related Enzyme Activities in Rats Fed with a High Fat and High Cholesterol Diet (황기홍국소스를 처리한 계육이 고지방 및 고콜레스테롤 식이 흰쥐의 ROS 생성 및 소거계 관련 효소의 활성에 미치는 영향)

  • Kim, Jae-Won;Kim, Soon-Dong;Youn, Kwang-Sup
    • Korean Journal of Food Science and Technology
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    • v.42 no.5
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    • pp.605-612
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    • 2010
  • The dietary effects of chicken treated with Hwangki-Beni Koji sauce (HBS) on reactive oxygen species (ROS) generating and scavenging related enzyme activities in rats fed with a high-fat and high-cholesterol diet were investigated. The rats (five rats per group) were divided into a normal control diet group (NC), a high-fat and high-cholesterol diet control group (HFC), HFC plus base sauce-treated chicken supplemented diet group (HFC-BS), and a HFC plus HBStreated chicken supplemented diet group (HFC-HBS), and fed for 5 weeks. Total type T (T) and type O (O) hepatic xanthine oxidoreductase in HFC-HBS were 27.91-35.78% and 24.57-31.84% lower than those of HFC and HFC-BS, respectively. In HFC-HBS compared with HFC and HFC-BS, superoxidase dismutase activity was 62.89-64.50% higher, glutathione S-transferase activity was 19.29-25.17% higher, glutathione content was 25.11-53.30% higher, and lipid peroxide content was 20.29-24.19% lower. Therefore, chicken treated with HBS may prevent liver damage by the ROS formed from a high-fat and high -cholesterol diet.

Inhibitory Effect of Spermidine with Antioxidant Activity on Oxidative Stress in Human Dermal Fibroblasts (사람피부섬유아세포에서 산화적 스트레스에 대한 항산화 활성을 가진 spermidine의 억제효과)

  • Park, In-Hwan;Kim, Moon-Moo
    • Journal of Life Science
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    • v.21 no.5
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    • pp.693-699
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    • 2011
  • Spermidine is a ubiquitous polycation that is synthesized from putrescine, which serves as a precursor of spermine. In recent years, spermidine was found to be a polyamine that plays an important role in longevity. Reactive oxygen species (ROS) such as hydroxyl radical, superoxide and hydrogen peroxide have been shown to be involved in various pathogenic processes as well as aging. The direct scavenging effect of spermidine on DPPH radical, $H_2O_2$ and hydroxyl radical, and its protective effect against DNA oxidation related to oxidative stress were evaluated in vitro. It was observed that spermidine exhibits scavenging activities on DPPH radical and H2O2 above 500 ${\mu}M$. Spermidine was especially effective in exerting a scavenging activity on hydroxyl radical. In addition, spermidine at 1000 ${\mu}M$ showed a clear protective effect against DNA oxidation. Furthermore, the expression level of anti-oxidant enzymes such as superoxide dismutase in humam dermal fibroblasts increased in the presence of spermidine compared with blank group. These results suggest that spermidine can be used as an antioxidant to prevent ROS-related diseases including inflammation, cancer and aging.

The Responses of Antioxidative Enzymes and Salt Tolerance of Atriplex gmelini (Atriplex gmelini(가는갯능쟁이)의 내염성과 항산화 효소 반응)

  • 배정진;윤호성;추연식;송승달
    • The Korean Journal of Ecology
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    • v.26 no.5
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    • pp.273-280
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    • 2003
  • Saline conditions invoke oxidative stress attributed to the overproduction of reactive oxygen species (ROS). Changes in quantum efficiency and antioxidative enzyme activity upon salt treatment were examined in a salt-tolerant plant, Atriplex gmelini, to test the hypothesis that salt tolerance of A. gmelini is due to the increased activity of antioxidative enzymes. A. gmelini showed optimum growth at 100 mM NaCl producing 116% of the shoot dry weight over control plants in 0 mM NaCl treatment. Healthy growth persisted up to 300 mM NaCl treatment maintaining normal internal water content and dry weight. No photochemical stress or damages on antioxidative defense system was obvious in plants of 2 and 4 day salt treatment which was indicated by increased quantum efficiency (Fv/Fm value), decreased stress index (Fo/Fm value), and increased activity of antioxidative enzymes such as SOD, APX, GR. However, the plants treated with 400 mM NaCl showed decrease in growth and in antioxidative enzyme activity although the enzyme activity was still higher than that of the 0 mM NaCl treated plants (l31%, 114%, and 134% of the SOD, APX, and GR activity, respectively). Interestingly, another important antioridative enzyme that scavenges H₂O₂ in plant cells, CAT, showed rapid decrease in its activity as salt concentration increased; 38%, 22%, 15% of the 0 mM NaCl treated plants at 200, 300, 400 mM NaCl treatments, respectively. It appears that the enzymes in ascorbate-glutathione cycle such as APX and GR play the major roles in scavenging ROS produced by salt stress in A. gmelini. After 6 days of salt treatment, the damage in photochemical and antioxidative defense system was indicated by decreased Fv/Fm value and increased Fo/Fm value. A. gmelini appears to cope with short term salt treatment by enhanced activity of the antioxidative defense system, whereas long term stress invoke oxidative stress by increased ROS due to the damages in photochemical and antioxidative system.

Effect of Scutellaria barbata Pharmacopuncture Extract on Degranulation and Inflammatory Mediator Release in RBL-2H3 Cells (반지련 약침 추출물의 RBL-2H3 세포 탈과립과 염증매개물질 분비 억제 효과)

  • Kwon, Hyuk-Sang;Song, Choon-Ho
    • Korean Journal of Acupuncture
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    • v.29 no.3
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    • pp.406-420
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    • 2012
  • Objectives : Scutellaria barbata has been widely used in oriental medicine used for treatment of acute and chronic inflammatory diseases. In this study, to investigate the protective effect of Scutellaria barbata on type I allergic response, we determined whether Scutellaria barbata inhibits early or late allergic responses. Methods : To assess the effect of Scutellaria barbata Pharmacopuncture Extract(SB) in RBL-2H3 cells, we investigated the levels of the markers of degranulation such as ${\beta}$-hexosaminidase and histamine, inflammatory mediator such as IL-4, TNF-${\alpha}$, PGE2 and cysLT, and mRNA expression of cytokines and enzymes. In addition, we determined the levels of intracellular ROS by DCFH-DA assay and the free radical scavenging activity by DPPH method. Results : We found that SB suppressed the release of ${\beta}$-hexosaminidase and histamine and the production of IL-4, TNF-${\alpha}$, PGE2 and cysLT in RBL-2H3 by the antigen stimulation. SB also significantly inhibited the enzyme mRNA expressions, such as HDC2, COX-1, COX-2, 5-LOX and iNOS2, along with reduced cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, SB suppressed the levels of intracellular ROS. Conclusions : Our results indicate that SB protects against type I allergic response and exert an anti-inflammatory effect through the inhibition of degranulation, inflammatory mediator release and mRNA expression of cytokines and enzymes.

Radioprotective effects of delphinidin on normal human lung cells against proton beam exposure

  • Kim, Hyun Mi;Kim, Suk Hee;Kang, Bo Sun
    • Nutrition Research and Practice
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    • v.12 no.1
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    • pp.41-46
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    • 2018
  • BACKGROUND/OBJECTIVES: Exposure of the normal lung tissue around the cancerous tumor during radiotherapy causes serious side effects such as pneumonitis and pulmonary fibrosis. Radioprotectors used during cancer radiotherapy could protect the patient from side effects induced by radiation injury of the normal tissue. Delphinidin has strong antioxidant properties, and it works as the driving force of a radioprotective effect by scavenging radiation-induced reactive oxygen species (ROS). However, no studies have been conducted on the radioprotective effect of delphinidin against high linear energy transfer radiation. Therefore, this study was undertaken to evaluate the radioprotective effects of delphinidin on human lung cells against a proton beam. MATERIALS/METHODS: Normal human lung cells (HEL 299 cells) were used for in vitro experiments. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay assessed the cytotoxicity of delphinidin and cell viability. The expression of radiation induced cellular ROS was measured by the 2'-7'-dicholordihydrofluorescein diacetate assay. Superoxide dismutase activity assay and catalase activity assay were used for evaluating the activity of corresponding enzymes. In addition, radioprotective effects on DNA damage-induced cellular apoptosis were evaluated by Western blot assay. RESULTS: Experimental analysis, including cell survival assay, MTT assay, and Western blot assay, revealed the radioprotective effects of delphinidin. These include restoring the activities of antioxidant enzymes of damaged cells, increase in the levels of pro-survival protein, and decrease of pro-apoptosis proteins. The results from different experiments were compatible with each to provide a substantial conclusion. CONCLUSION: Low concentration ($2.5{\mu}M/mL$) of delphinidin administration prior to radiation exposure was radioprotective against a low dose of proton beam exposure. Hence, delphinidin is a promising shielding agent against radiation, protecting the normal tissues around a cancerous tumor, which are unintentionally exposed to low doses of radiation during proton therapy.