• Title/Summary/Keyword: Reductase

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Antioxidant Activities of Hot Water Extract from Cornus walteri Wanger against Oxidative Stress Induced by tert-Butyl Hydroperoxide in HepG2 Cells (tert-Butyl Hydroperoxide로 산화 스트레스가 유도된 HepG2 세포에서 말채나무 열수추출물의 항산화 활성)

  • Yeon, Seong Ho;Ham, Hyeonmi;Sung, Jeehye;Kim, Younghwa;Namkoong, Seulgi;Jeong, Heon-Sang;Lee, Junsoo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.10
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    • pp.1525-1532
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    • 2013
  • The objective of this study was to investigate the effect of hot water extract from Cornus walteri Wanger (CWE) on tert-butyl hydroperoxide (TBHP)-induced oxidative stress in HepG2 cells. Generation of reactive oxygen species (ROS), concentrations of cellular lipid peroxidation products and reduced glutathione, and antioxidant enzyme activity were used as biomakers of cellular oxidative status. Cells pretreated with CWE (25~200 ${\mu}g/mL$) showed an increased resistance to oxidative stress in a dose-dependent manner, as revealed by a higher percentage of surviving cells compared to control cells. ROS generation induced by TBHP was significantly reduced when cells were pretreated with 200 ${\mu}g/mL$ CWE for 4 h. Pretreatment with CWE (5~50 ${\mu}g/mL$) prevented the decrease in reduced glutathione and the increase in malondialdehyde and ROS evoked by TBHP in HepG2 cells. Finally, CWE pretreatments prevented the significant increase of glutathione peroxidase, catalase, glutathione reductase, and superoxide dismutase activities induced by TBHP. These results show that CWE has significant protective ability against a TBHP-induced oxidative insult and that the modulation of antioxidant enzymes by CWE may have an important antioxidant effect on TBHP-induced oxidative stress in HepG2 cells.

Molecular cloning and expression analysis of the first two key genes through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway from Pyropia haitanensis (Bangiales, Rhodophyta)

  • Du, Yu;Guan, Jian;Xu, Ruijun;Liu, Xin;Shen, Weijie;Ma, Yafeng;He, Yuan;Shen, Songdong
    • ALGAE
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    • v.32 no.4
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    • pp.359-377
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    • 2017
  • Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially useful macroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically important metabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway were cloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete open reading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showed that PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibited by fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higher plants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in line with the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only complete MEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivatives through mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. This study demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to research the problem of chromalveolata biological evolution.

Effects of Mori Fructus Extract on Hyperglycemia, Hyperlipidemia, Polyol Pathway, AGE and RAGE in ob/ob Mice (상침자(桑椹子)추출물이 ob/ob mouse의 혈당, 혈청 지질, Polyol Pathway, AGE 및 RAGE에 미치는 영향)

  • Lee, Min-Dong;Jeong, Ji-Cheon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.2
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    • pp.351-359
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    • 2009
  • Etiological studies of diabetes and its complications showed that oxidative stress might play a major role, Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Mori Fructus extract has been known to be effective for the antidiabetic, antihyperlipidemic and antiobesitic prescription, and composed of four crude herbs, The aim of this study was to investigate the effect of Mori Fructus extract in male ob/ob mouse with severe obesity, hyperinsulinemia, hyperglycemia, hyperlipidemia. Mice were grouped and treated for 5 weeks as follows. Both the lean (C57BL/6J black mice) and diabetic (ob/ob mice) control groups received standard chow. The experimental groups were fed with a diet of chow supplemented with 7.5, 15 and 30 mg Mori Fructus extract per 1 kg of body weight for 14 days. The effects of Mori Fructus extract on the ob/ob mice were observed by measuring the serum levels of glucose, insulin, lipid components, and the kidney levels of reactive oxygen species (ROS), MDA+HAE, GSH and also the enzyme activities involved in polyol pathway. Western blotting was performed using anti-AGE, anti-RAGE respectively. Mori Fructus extract lowered the levels of serum glucose and insulin in a dose dependent manner. Total cholesterol, triglyceride and free fatty acid levels were decreased, while the HDL-cholesterol level was increased, in Mori Fructus extract treated groups. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the ob/ob mice, whereas those were inhibited in the Mori Fructus extract-administered groups. Mori Fructus extract inhibited the generation of ROS in the kidney. MDA+HAE level was increased and the GSH level was decreased in the ob/ob mice, whereas those were improved in the Mori Fructus extract-administered groups. Mori Fructus extract inhibited the expression of AGE, RAGE in the ob/ob mice. The results suggested that Mori Fructus exerted the antidiabetic and antihyperlipidemic activities by regulating the activities of polyol pathway enzymes, scavenging the ROS, decreasing the MDA+HAE level, increasing the GSH level and inhibiting the expression of AGE, RAGE in the ob/ob mice.

Comparative Analysis of Local Green Tea in Korea by STS-RFLP (STS-RFLP법을 이용한 국내지역 재배녹차의 비교분석)

  • Cho, Kiu-Hyung;Jo, Aruna;Tsuge, Tomohiko;Kim, Jong-Cheol;Kim, Ru-Mi;Yoon, Ho-Sung;Kim, Gyung-Tae
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1415-1419
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    • 2010
  • Consumption of green tea has increased along with increasing concern regarding healthier lifestyles, and many brands of green tea are sold with a label indicating the region of Korea in which the tea was produced. However, there is little information on identifying the difference between the green tea cultivars according to the region they were grown. Here, 9 green tea cultivars collected from Hadong region, Bosung region, China and Japan were subjected to the STS-RFLP analysis. Using the coding and noncoding DNA regions of genes related to the phenylpropanoid pathway, such as phenylalanine ammonia-lyase, chalcone synthase and dihydroflavonol 4-reductase, we have identified the differences between green tea cultivars according to the region they were grown in. In this study, we showed a STS-RFLP method of green tea analysis which easily distinguished different kinds of tea using the primers as described. In addition, we identified that the green tea cultivars from Hadong and Bosung displayed a different profile when PAL intron was digested with Dde I, suggesting that a rapid authentication system for green tea cultivars grown in different regions in Korea is available.

Metabolism of Lactate Dehydrogenase in Tissues from Ldh-C Expressed Mice (Mus musculus) in a Starvation State (기아상태에서 Ldh-C가 발현된 생쥐(Mus musculus) 조직의 젖산탈수소효소의 대사)

  • Yum, Jung Joo;Kim, Gyu Dong
    • Journal of Life Science
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    • v.28 no.1
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    • pp.1-8
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    • 2018
  • To confirm the function of lactate dehydrogenase (LDH) (EC 1.1.1.27, LDH), its metabolism was studied by activity, kinetics, and isozyme analysis in tissues of Ldh testis-specific C expressing mice (Mus musculus) maintained in a state of starvation for 48 hr and 96 hr. In skeletal muscle, liver, and eye tissues, LDH and LDH $A_4$ activity increased and anaerobic metabolism predominated. While LDH activity in the heart and kidney tissues decreased, LDH $B_4$ activity increased and aerobic metabolism predominated, producing pyruvic acid. In the testis tissue, LDH $C_4$ activity decreased. In the brain tissue, LDH activity increased, but the isozyme change was small and the amount of pyruvic acid decreased. $K{_m}^{PYR}$ increased in tissues other than kidney tissue, and the affinity for pyruvic acid decreased. Consequently, in Ldh-A and B-expressing tissues, the activities of isozymes with higher concentrations increased. However, in Ldh-A, B, and C-expressing tissue, $C_4$ decreased and the function of the tissue also decreased. In particular, LDH in brain tissue played a role as a pyruvate reductase. Therefore, this process might be the mechanism for producing energy in the state of starvation.

Changes of Growth and Antioxidative Enzyme(SOD, APX, GR) Activities of Spinach Beet(Beta vulgaris var. cicla) Under Saline Condition (염 환경하에서 근대(Beta vulgaris var. cicla)의 생장과 항산화효소(SOD, APX, GR)의 활성변화)

  • 배정진;추연식;송승달
    • Journal of Life Science
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    • v.13 no.5
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    • pp.658-667
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    • 2003
  • Antioxidative enzymes (superoxide dismutase; SOD, ascorbate peroxidase; APX, glutathione reductase; GR) play major roles in scavenging mechanism of reactive oxygen species which were involved in various stress conditions including salt. In order to investigate the relation between their growth responses (dry weight) and the changes of antioxidative enzymes activity, salt-tolerant spinach beet having 15cm of shoot length were treated with various salt levels (0, 50, 200, 1000 mM NaCl) for 24 hours. Spinach beet exhibited an increase in the activity of antioxidative enzymes by salt, the maximal activity at 200 mM NaCl and the lowest activity at 50 mM NaCl in 2 hrs. after treatments. As a result of PAGE, it has been confirmed that spinach beet contained 3 isoforms (Fe-SOD, CuZn-SOD and Mn-SOD) of SOD and main isoform was CuZn- SOD form. In case of APX, isoforms of the low molecular weight(No. 7, 8) were showed strong expression especially at 200 and 400 mM NaCl treatment. Meanwhile, GR did not show specific pattern of isoforms among the salt treatments. Especially, in case of 50 mM treatment, plant showed the lowest activity of SOD with the best growth, a low enzyme activity was induced by inactivation of the Mn-SOD. Therefore, we suggested that the decrease of SOD activity at a low salt level (50 mM NaCl) or the increase of enzyme activity at a high salt level (200 mM NaCl) may be related to expression of the Mn-SOD isoform. These antioxidative enzymes showed the increase of activity in a short time by salt addition. So, it is considered that spinach beet copes effectively with a stressful condition such as salt by operating effective antioxidative defense mechanism rapidly under high salt level.

Anti-Inflammatory and Antidiabetic Effects of Brown Rice (Oryza sativa L.) Extracts (항염증 및 항당뇨 활성에 미치는 현미 추출물의 영향)

  • Cho, Eun-Kyung;Jung, Kyung-Im;Choi, Young-Ju
    • Journal of Life Science
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    • v.22 no.1
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    • pp.126-131
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    • 2012
  • Physiological activities of hot water (BRW) and 80% ethanol (BRE) extracts from brown rice were investigated in this study. The highest activity (94.9%) of nitrite reductase was observed for BRE at 1 mg/ml at pH 1.2, while the activity for BRW was about 75.4% under the same conditions. The inhibitory effects of BRW and BRE on xanthine oxidase activity were about 39.0 and 72.9% at 10 mg/ml, respectively. The digestibility of starch was lower for brown rice than for milled rice and the highest inhibition (93.1%) of ${\alpha}$-glucosidase activity occurred with BRE. Superoxide dismutase (SOD)-like activities of BRW and BRE were weakly increased in a dose-dependent manner and were about 56.4 and 44.9% at 10 mg/ml, respectively. The influences of BRW and BRE on alcohol metabolizing activity were determined by measuring the generation of reduced nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). Increases in ADH and ALDH activities were only detected with BRE.

Expression of ERCC1, RRM1 and LRP in Non-small Cell Lung Cancers and their Influence on Chemotherapeutic Efficacy of Gemcitabine Concomitant with Nedaplatin

  • Qiu, Zhen-Qin;Zhao, Kun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7303-7307
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    • 2014
  • Objective: To explore the clinical efficacy of gemcitabine concomitant with nedaplatin and drug resistance in the treatment of non-small cell lung cancer (NSCLC) and associated molecular predicators. Materials and Methods: A total of 68 patients diagnosed with NSCLC by histology served as the study objects and were randomly divided into an observation group treated with gemcitabine concomitant with nedaplatin and a control group with cisplatin concomitant with gemcitabine, 34 cases for each group. Short-term and long-term efficacies, adverse responses as well as the expression of nucleotide excision repair cross complementing 1 (ERCC1), ribonucleotide reductase subunit M1 (RRM1) and lung resistance-related protein (LRP) in NSCLC tissues in both groups were assessed. Results: The short-term objective response rate (ORR) and disease control rate (DCR) were 35.3% (12/34) and 76.5% (26/34) in the observation group and 38.2% (13/34) and 85.3% (29/34) in the control group, respectively, the differences not being statistically significant. The time to progression (TTP) in both groups were 1~12 months, while the median TTP was 135 d and 144 d, respectively. Though the survival was slightly higher in the control group, there were no significant differences in TTP and survival time. The rates of decreased hemoglobin, vomiting and nausea as well as renal toxicity were evidently lower in the observation group, while other adverse responses demonstrated no significant difference. The positive expression rates of ERCC1, RRM1 and LRP were 47.1% (16/34), 61.8% (21/34) and 64.7% (22/34) in the observation group, respectively. Compared with negative ERCC1 expression, ORR had decreasing trend and the overall survival time (OS) decreased significantly in patients with positive ERCC1 expression, which were markedly decreased by the positive expressions of RRM1 and LRP. Conclusions: Gemcitabine concomitant with nedaplatin has significant effects in the treatment of NSCLC, with an adverse response rate obviously lower than for cisplatin concomitant with gemcitabine, suggesting that wider use in the clinic is warranted. Additionally, the positive expressions of ERCC1, RRM1 and LRP may increase patient drug resistance, so they can be applied as the chemotherapeutic predicators to guide individualized therapy of NSCLC patients.

The MTHFR C677T Polymorphism and Risk of Acute Lymphoblastic Leukemia: an Updated Meta-analysis Based on 37 Case-control Studies

  • Jiang, Yuan;Hou, Jing;Zhang, Qiang;Jia, Shu-Ting;Wang, Bo-Yuan;Zhang, Ji-Hong;Tang, Wen-Ru;Luo, Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6357-6362
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    • 2013
  • Background: The C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) has been associated with acute lymphoblastic leukemia (ALL). However, results were conflicting. The aim of this study was to quantitatively summarize the evidence for the MTHFRC677T polymorphism and ALL risk. Methods: Electronic searches of PubMed and the Chinese Biomedicine database were conducted to select case-control studies containing available genotype frequencies of C677T and the odds ratio (OR) with 95% confidence interval (CI) was used to assess the strength of any association. Results: Case-control studies including 6,371 cases and 10,850 controls were identified. The meta-analysis stratified by ethnicity showed that individuals with the homozygous TT genotype had decreased risk of ALL (OR= 0.776, 95% CI: 0.687~0.877, p< 0.001) in Caucasians (OR= 0.715, 95% CI: 0.655~0.781, p= 0.000). However, results among Asians (OR=0.711, 95% CI: 0.591~1.005, p= 0.055) and others (OR=0.913, 95% CI: 0.656~1.271, p= 0. 590) did not suggest an association. A symmetric funnel plot, the Egger's test (P=0.093), and the Begg- test (P=0.072) were all suggestive of the lack of publication bias. Conclusion: This meta-analysis supports the idea that the MTHFR C677T genotype is associated with risk of ALL in Caucasians. To draw comprehensive and true conclusions, further prospective studies with larger numbers of participants worldwide are needed to examine associations between the MTHFRC677T polymorphism and ALL.

Enzyme-Histochemical Study of Philtral Orbicularis Oris Muscle Fiber Types in Korean Male Cadaver (한국인 성인남성 사체에서 시행한 인중 구륜근 섬유들의 효소-조직화학적 분석)

  • Yu, Myung-Sook;Park, Jung-Min;Lee, Hee-Su;Lee, Suk-Keun;Kang, Ji-Young;Eo, Mi-Young;Lee, Jong-Ho;Kim, Soung-Min
    • Korean Journal of Cleft Lip And Palate
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    • v.12 no.2
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    • pp.47-56
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    • 2009
  • The orbicularis oris muscle (OOM) is a very important muscle that originate from the second branchial arch and is innervated by the facial nerve. The aim of this study was to elucidate distribution types of two muscle fibers that composing OOM by using enzyme-histochemical examinations and tried to make a basis for a clinical application. The fresh frozen tissues from the superior and inferior portions of the OOM were taken from post mortem 65-year-old Korean male adult. Total five different sagittal sections were used on the midline of the philtrum, the middle portion of lower lip, the mouth corner, and each midlateral side of upper and lower mouth. We used enzyme-histochemical staining such as Periodic Acid-Schiff (PAS), Succinic Dehydrogenase (SDHase), reduced Nicotinamide Adenine Dinucleotide-Tetrazolium Reductase (NADH-TR), Adenosine Triphosphatase (ATPase) in pH 9.4, 4.6 and 4.3, and Modified Gomori Trichrome. There were about 30.24 % type 1 muscle fiber and 65.40 % type 2 muscle fiber in the midline of the philtrum (p < 0.05). Enzyme-histochemical staining is very useful and innovative method to elucidate characteristics of muscle fibers. We expect that chiloplasty and reconstruction of the lip portions for cleft lip patients, based on these results, are better to recovery function and aesthetic. However, we have some problems as an intramuscular variability and the inter-individual variation etc. Therefore we have to make progress these studies continuously to overcome these problems.

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