• Title/Summary/Keyword: LDH activity assay

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Neuroprotective Effect according to Reactive Oxygen Species Scavenging Activity from Extracts of Cudrania tricuspidata Leaves (활성산소 소거활성에 따른 꾸지뽕잎 추출물의 신경세포 보호 효과)

  • Kang, Young-Kyoung;Lee, Eun-Ah;Park, Hae-Ryong
    • Korean journal of food and cookery science
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    • v.28 no.6
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    • pp.821-828
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    • 2012
  • In an attempt to identify the neuroprotective effect of Cudrania tricuspidata (CT) leaves against ROS (reactive oxygen species)-induced oxidative stress in neuronal cells, the extracts from CT leaves were investigated using PC12 cells and N18-RE-105 cells. The methanolic and ethanolic extracts from CT were denoted as CTM (Cudrania tricuspidata Leaves methanolic extracts) and CTE (Cudrania tricuspidata Leaves ethanolic extracts), respectively. The neuroprotective effects of the extracts were measured by DCF-DA assay, MTT reduction assay, and LDH release assay. The PC12 cells exposed to $H_2O_2$-induced oxidative stress and the N18-RE-105 cells exposed to glutamate-induced oxidative stress were treated with various concentrations of CTM and CTE. The results, CTM treatments resulted in the induction of a dose-dependent protective effect in PC12 cells and N18-RE-105 cells. Interestingly, CTE also showed neuroprotective effect in PC12 cells and N18-RE-105 cells. Therefore, these results suggest that CTM and CTE could be a new potential candidate as neuroprotective agents against ROS-induced oxidative stress in neuronal cells.

Effects of Taurine on Glutamate-induced Neurotoxicity and Interleukin-6 mRNA Expression in Astrocytes

  • Yang, Seong-Chil;Baek, Su-Yeon;Choe, In-Pyo;Lee, Chang-Jung
    • Animal cells and systems
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    • v.1 no.3
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    • pp.467-473
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    • 1997
  • Taurine (2-aminoethanesulfonic acid), one of bioactive amino acid in the mammalian brain, is known to exert inhibitory effects on neurons via GABA receptor. In the present study, we examined effects of taurine on glutamateinduced neurotoxicity on hippocampal neuron cell culture using cell counting method and lactate dehydrogenase (LDH) assay. After 10 d of culture, cells were stimulated with appropriate drugs. Only 43% of cultured neuronal cells survived at one day after stimulation with 500 uM L-glutamate for 10 min. Survival rate was enhanced by 82% in the presence of 10 mM taurine. LDH activity from the culture supernatant incubated with a combination of L-glutamate and taurine was less than half of that with L-glutamate alone. In the next series of experiments, interleukin-6 (IL-6) mRNA expression in cultured astrocytes was investigated using reverse tanscription-PCR (RT-PCR). IL-6 mRNA was detected in the astrocytes stimulated with L-glutamate in a dose-dependent manner, while not detected in the unstimulated control astrocytes. The expression of IL-6 mRNA caused by 10 mM glutamate was inhibited by taurine, but not by GABA. These findings demonstrated a neuroprotective action of taurine against glutamate-induced toxicity.

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The effect of Holotrichia on Natural killer Cell Activity in Mice (제조가 마우스 Natural killer 세포(細胞) 활성(活性)에 미치는 영향(影響))

  • Kim, Gi-Yeol;Kim, Jong-Dae;Jeong, Ji-Cheon;Nam, Kyung-Soo
    • The Journal of Korean Medicine
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    • v.19 no.2
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    • pp.313-325
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    • 1998
  • The effect of Holotrichia on natural killer cell activity in normal mouse were studied. 1. The oral administration of Holotrichia increased spleen weight about 21.1% and also cell numbers of spleen compared to control mice group. 2. The cytotoxicity of effector cell was most effectively induced in a ratio of 50 : 1(effector/target cell). 3. Cytotoxicity of effector cells was. increased about 24% as compared with control group in in vivo test. 4. On the other hand, the administration of Holotrichia original solution showed significant increase the cytotoxicity. The cytotoxicity was increased concentration dependently. 5. The cytotoxicity by $^{3}H-thymidine$ incorporation assay showed similar effect with LDH enzyme method. 6. In the purified NK cells, the cytotoxicity was increased about 31% as compared with control group in in vivo system and the ratio of cytotoxicity was generally more increased than that of partially purified NK cell. 7. In vitro experimet of the purified NK cells, the cytotoxicity was increased 11.8% as compared with control group and the ratio of cytotoxicity was also more increased than that of partially purified NK cell. These results suggest that Holotrichia is administrated to mice with malignant tumors, the increase of NK cell activity may occur and affect tumor cells.

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Evaluation of the Antioxidant and Antiproliferative Properties of a Hot-water Extract from Gulfweed, Sargassum fulvellum

  • Kim, So Jung;Kang, Mingyeong;Lee, Taek-Kyun
    • Journal of Marine Bioscience and Biotechnology
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    • v.10 no.2
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    • pp.53-61
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    • 2018
  • Sargassum fulvellum (gulfweed) is a widespread seaweed in the coastal areas of northeast Asia. In the present study, we identified the phenolic compounds present in aqueous and ethanolic extracts of S. fulvellum and evaluated their antioxidative properties and their abilities to block cell proliferation using in vitro assays: antioxidant activity was assessed by using a DPPH assay and superoxide anion scavenging activity, anti-tyrosinase activity, and anti-proliferative activity were assessed using MTT and lactate dehydrogenase [LDH] assays in vascular smooth muscle cells. The hot-water ($65^{\circ}C$) extract had a higher phenol content than the ethanolic extract. The hot-water extract showed a statistically significant increase in free radical scavenging activity and a greater ability to reduce proliferation of vascular smooth muscle cells stimulated with platelet-derived growth factor-BB. Taken together, hot-water extracts of S. fulvellum may be an important source of antioxidative and antiproliferative agents.

Studies on the Mutagenicity and Hepatotoxicity of Antaeum (안태음의 변이원성 및 간독성에 관한 연구)

  • Lee, Dong-Nyung;Moon, Jin-Young;Oh, Gue-Suc;Lee, Tae-Kyun;Choi, Mi-Jung;Lee, Dong-Mok;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.28 no.3
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    • pp.149-155
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    • 1997
  • Antaeum(ATE) has been used as a prescription for threatened abortion, associated with pregnancy in traditional medicine. Because gravida could be administered ATE for a long period, its administration might cause a harmful effect on fetus and gravida during the pregnancy. This study aimed to determine whether exposure to ATE caused mutagenicity or hepatotoxicity during the pregnant period. For mutagenicity test of ATE, Salmonella typhimurium and Bacillus subtilis were used as indications for DNA damage. In the Ames test, Samonella typhimurium TA98 and TA100 were used for mutagenicity testing, and the number of histidine revertants was measured. In Rec-assay, Bacillus subtilis H $17(Rec^+)$ and $M-45(Rec^-)$ strains were used to clarify the DNA damage property. In the SOS umu test, Salmonella typhimurium TA15335 containing plasmid pSK1002 was used as a tester strain, and we monitored the levels of umu operon expression by measuring the ${\beta}-galactosidase$ activity. From the tested results, ATE did not show DNA damage and mutagenicity. On the other hand, hepatotoxicity of ATE to female ICR mice was monitored by the measurements of s-GOT, s-GPT and LDH activities after oral feeding for 15 days. ATE did not show significant change of s-GOT, s-GPT and LDH activities in mice sera.

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Antioxidant and Neuronal Cell Protective Effect of Purple Sweet Potato Extract (자색고구마 추출물의 항산화 효과 및 신경세포 보호효과)

  • Kwak, Ji-Hyun;Choi, Gwi-Nam;Park, Ju-Hee;Kim, Ji-Hye;Jeong, Hee-Rok;Jeong, Chang-Ho;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.44 no.2
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    • pp.57-66
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    • 2010
  • The antioxidant and neuronal cell protective effects of water extract from purple sweet potato were investigated. The total phenolics and monomeric anthocyanin contents of purple sweet potato extract were 44.25 mg/g and 2,394 mg/L, respectively. The antioxidant activities of purple sweet potato extract were evaluated using various antioxidant tests, including 1,1-diphenyl- 2-picrylhydrazyl (DPPH), 2,2'-azino- bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing/antioxidant power (FRAP) and reducing power. In these assays, the extract of purple sweet potato presented significant radical scavenging activities, FRAP, and reducing power in a dose-dependent manner. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl- tetrazoliumbromide} reduction assay showed significantly increase in cell viability when PC12 cells were pretreated with purple sweet potato extract. Because oxidative stress is also known to increase neuronal cell membrane breakdown, we further investigated by lactate dehydrogenase (LDH) and neutral red uptake assay. Purple sweet potato extract inhibited oxidative stress-induced membrane damage in neuronal cells. Therefore, these data results demonstrated that water extract of purple sweet potato have antioxidant activity and neuronal cell protective effect thus it has great potential as a natural source for human health.

Acute Pulmonary Responses in Vivo to Silica Complexed with $H^+$, $Zn^{2+}$, or $Fe^{3+}$

  • Lee, Ji-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.2
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    • pp.183-189
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    • 1999
  • This investigation is to determine whether the surface complexation of iron influence acute pulmonary responses induced by silica. For this study, three varieties of cation complexed silica were used: $silica-H^+,\;-Zn^{2+},\;and\;-Fe^{3+},$ since the first two are not active in the transport of electrons and generate little free radicals relative to the dust with the surface iron. Rats (270 to 280 g) were intratracheally (IT) instilled with saline, $silica-H^+,\;-Zn^{2+},\;or\;-Fe^{3+}$(5 mg in 0.5 ml saline). After 4 h, cell number, type, and differentiation were analysed in the bronchoalveolar lavage cells, and the levels of lactate dehydrogenase (LDH) and total protein were determined in the lavage fluid. In addition, bronchoalveolar lavage cells were cultured, and nitric oxide production was measured using nitrate assay. Inducible nitric oxide synthase (iNOS) mRNA in the bronchoalveolar lavage cells was also determined by northern blot analysis. Differential counts of the lavage cells showed that red blood cells were increased by 9-, 8-, and 13-fold and total leukocytes (lymphocytes plus polymorphonuclear neutrophils) by 48-, 36-, and 33-fold, following IT $silica-H^+,\;-Zn^{2+},\;and\;-Fe^{3+},$ respectively compared with the saline group. Meanwhile, there were no significant differences in red blood cells and total leukocytes among any of the cation complexed silica groups. The levels of LDH and total protein in the lavage fluid were significantly increased by 3- to 4-fold. However, compared among these silica groups, $Fe^{3+}$? complexation did not significantly change the LDH activity and total protein. NO production in cultured bronchoalveolar lavage cells was elevated by 2-fold, following IT any of the silica treatments compared with the saline group. Furthermore, the steady-state levels of iNOS mRNA in the lavage cells were greatly increased. There were any differences in iNOS mRNA expression among the silica-treated groups as with NO production. These findings suggest that surface complexed iron may not influence the acute pulmonary responses resulted from 4h exposure to silica.

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Effects of Sophorae Radix Water Extract on Cultured Rat Myocardial Cells (고삼(苦蔘) 전탕액(煎湯液)이 배양심근세포(培養心筋細胞)에 미치는 영향(影響))

  • Kim, Hyun-Kyu;Park, Jun-Su;Kwon, Kang-Beom;Lee, Ho-Sub;Han, Jong-Hyun;Park, Seung-Taeck;Ryu, Do-Gon
    • The Journal of Korean Medicine
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    • v.20 no.1 s.37
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    • pp.142-150
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    • 1999
  • In order to elucidate toxic the mechanism of myocardial damage and the protective effect of herbal extract, Sophorae Radix(SR) against myocardiotoxicity, the cytotoxic effect of adriamycin and cardioprotective effect of SR were examined by MTT assay, LDH activity, heart beat rate and light microscopy after cultured myocardial cells derived from neonatal mouse were treated with various concentrations of adriamycin, an inducer of myocardiotoxicity. Adriamycin induced a decrease of cell viability, an increase in the amount of lactate dehydrogenase(LDH), and a decrease in the heart beat rate and a decrease in the number of cells, when administered to cultures myocardial cells in a dose-dependent manner. In cardioprotective effect of SR. SR showed the decrease of amount of LDH, and an increase of heart beating rate and cells in number on cultured myocardial cells damaged by adriamycin. From the above results, it is suggested that adriamycin shows toxic effect in cultured myocardial cells derived from a neonatal mouse, and herbal extract such as SR is very effective in the prevention of adriamycin-induced cardiotoxicity.

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Antioxidant Effects of Eriodictyol on Hydrogen Peroxide-Induced Oxidative Stress in HepG2 Cells (산화스트레스가 유도된 HepG2 세포에서 Eriodictyol의 항산화 효과)

  • Joo, Tae-Woo;Hong, Sung-Hyun;Park, Sun-Young;Kim, Gur-Yoo;Jhoo, Jin-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.510-517
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    • 2016
  • This study was conducted to investigate the antioxidant and hepatoprotective effects of eriodictyol compound against hydrogen peroxide-induced oxidative stress in HepG2 cells by measuring expression levels of antioxidant enzymes, liver function index enzyme activities, and inhibitory effects against reactive oxygen species (ROS) production. HepG2 cell viability was assessed using 3-(4,5-dimethyl thiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. In the concentration range of $10{\sim}50{\mu}g/mL$, eriodictyol displayed over 98% cell viability in HepG2 cells. The effects of increased gene expression on hydrogen peroxide-induced oxidative stress were analyzed by monitoring antioxidant enzyme (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPx) gene expression levels using real-time PCR. Eriodictyol compound significantly increased gene expression levels of SOD, CAT, and GPx in a dose-dependent manner ($10{\sim}50{\mu}g/mL$). Hepatoprotective effects against hydrogen peroxide-induced oxidative stress were analyzed by monitoring glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) activities in HepG2 cell culture medium using a biochemistry analyzer. Eriodictyol compound significantly reduced GOT, LDH, and GGT activities in a dose-dependent manner in HepG2 cells. ROS level in HepG2 cells was analyzed by 2',7'-dichlorofluorescein fluorescence diacetate assay, and eriodictyol compound effectively reduced the intracellular ROS level in HepG2 cells. The results reveal that eriodictyol compound can be useful for development of effective antioxidant and hepatoprotective agents.

Localization and characteristics of lactate and palate dehydrogenase in the sparganum and adult worm of Spirometra erinacei (만손열두조충에서 젖산탈수소효소와 말산탈수소효소의 분포와 특징)

  • Gwak, Gi-Hun;Jeon, Eun-U;Kim, Chang-Hwan
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.59-68
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    • 1996
  • This study investigated the enzyme histochemical localization and characteristics of lactate (LDH) and malate dehydrogenase (MDH) related with the oxidation-reduction metabolism in the sparganum and adult of 5. erinacei. By enzyme histochemical assay, activity of LDH was strong in the tegument and subtegumental muscle layers of the adult and sparganum. Activity of MDH was strong in the tegument of the sparganum and subtegumental muscle layers of the adult. However it was weak in the tegument of the adult. By electrophoresis, 45 kDa band was major and common in LDH of adults and spargana. The 150 kDa molecule was the major and common band in MDH of adults and r -spargana (from experimentally infected rats) . By isoelectrofocusing, isoelectric points (Pl) or 4 MDH isogyme from adult worm were 6.0.6.5, 6.7 and 7.1, respectively. Pl 6.0 was the major band. The active range of pH for MDH was about pH 6-8 and the optimum pH was pH 7 The effective temperature on the MDH was about $30^{\circ}C$$50^{\circ}C$ and the optimum temperature was about 40℃ in spargana md adult worm. In the stability against heat, when MDH was heated at 85℃ for 10 seconds, the activity was denatured perfectly. Maximum activity or MDH was 19.4 unit in the s-sparganum (from snakes), 24.5 unit in the r-sparganum (from rats) and 108.0 unit in the adult worm. The maximum activity was higher in adults than in spargana. The present result showed us that the nutrients absorbed through the tegument were transferred into inner tissues and were utilized as the source of metabolism. According to the habitat of the parasite, the isozymes of LDH and MDH are activated differently, and by this different activation the sparganum and adult can adapt themselves to parasitic circumstances.

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