• The Korean Journal of Zoology
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• v.12 no.3
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• pp.94-102
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• 1969

The protective action of methylene blue against gamma-irradiation was studied with rats. Albino rats were given 360 rads of whole-body gamma-irradiation following an intraperitoneal injection of physiological saline or methylene blue. Male rats given methylene blue (38mg/kg) and the control rats given saline were alive following gamma-irradiation. Serum lactic dehydrogenase (LDH) activity, and LDH isoenzyme patterns in serum and various organs were determined at various time intervals after the exposure. 1) The serum LDH level in both the control and methylene blue-treated rats was increased during the initial phase, but returned to the initial level thereafter. 2) Methylene blue showed a marked delay in the rise of serum LDH at 15 and 64 hours after exposure. 3) The exposure in the control and methylene blue-treated rats resulted in an increase in the relative amount of the more electrophoretically mobile-anodal isoenzyme (band 1) and a decrease in the least mobile-cathodal isoenzyme (band 5) in serum, liver, heart and testis nearly at 40 and 116 hours, respectively. 4) Isoenzyme patterns in serum, liver and testis after exposure were not significantly different between the control and the methylene blue-treated rats. 5) Methylene blue showed a slight delay in alteration of heart tissue LDH isoenzyme patterns after exposure. 6) The increase of serum LDH level after exposure is a reflection of an immediate increase in the H type, band 1 of LDH isoenzymes. 7) It is concluded from this study that methylene blue has a remarkable radioprotective action in the serum LDH activity and in the heart tissue LDH isoenzyme patterns.

• The Journal of Korean Medicine
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• v.21 no.2
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• pp.79-86
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• 2000

Objectives and Methods : In order to elucidate toxic mechanism of myocardial damage and protective effect of myrrha water extract against cytotoxic effect of xanthine oxidase/hypoxanthine(XO/HX), cardioprotective effect of myrrha water extract was examined by MTT assay, LDH (Lactate Dehydrogenase) activity and heart beating rate after cultured myocardial cells derived from neonatal mouse were treated with various concentration of XO/HX, a free radical. Results : XO/HX induced a decrease of cell viability, an increase in the amount of LDH, and a decrease of heart beating rate on cultured myocardial cells in a dose-dependent manner. In cardioprotective effect of myrrha water extract, it showed a decrease in the amount of LDH and an increase of heart beating rate on cultured myocardial cells damaged by XO/HX. Conclusions : From the above results, it is suggested that XO/HX showed toxic effect in cultured myocardial cells derived from neonatal mouse and that myrrha water extract is very effective in the prevention of XO/HX-induced cardiotoxicity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.4
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• pp.996-1001
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• 2003

It is well known that oxidative stress of reactive oxygen species(ROS) may be a causative factor in the pathogenesis of bone disorder. The purpose of this study was to evaluate the cytotoxicity of oxidative stress. Cell viability by MTS assay or INT assay, activity of glutathione peroxidase(GPx), lipid peroxidation(LPO) activity and cell viablity. And also protctive effect of glutamate receptors against ROS-induced osteotoxicity was examined by protein synthesis, alkaline phosphatase (ALP) activity and lactate dehydrogenase (LDH) activity in cultured rat osteoblasts and osteoclasts. XO/HX decreased cell viability and GPx activity, protein synthesis and ALP activity, but increased LPO activity and LDH activity. In the protective effect, N-methyl-D-aspartate (NMDA) receptor antagonists or AMPA/kainate receptor antagonists such as D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), NMDA receptor antagonists but AMPA/kainate receptor antagonists showed protective effect on xanthine oxidase (XO) and hypoxanthine (HX) in these cultures by the increse of protein synthesis, ALP activity.

• Applied Biological Chemistry
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• v.36 no.1
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• pp.64-69
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• 1993

This study was performed to investigate detoxifying enzyme activities of carboxylesterase(CE), glutathione S-transferase(GST) and lactate dehydrogenase(LDH) at variable toxicity levels in fresh water fish, carp(Cyprinus carpio L.). The carp was exposed to single and combined pesticides of IBP, isoprothiolane and cartap for 48 hr at sublethal doses, $LC_{10}$ and $LC_{26}$. The detoxifying enzyme activities were assayed for the liver, head and gut of the carp. The enzyme activities we discovered were as follows: Both activities of CE and GST were increased at the sublethal doses but were declined by increasing doses. In the gut, we found that the CE activity had high levels in the treatment groups of isoprothiolane+IBP and isoprothiolane+cartap. In the head, the CE activity had high levels in the treatment groups of cartap, IBP and isoprothiolane. However, the GST activities were inconsistent in the head and gut of the fish. Also, the GST activity was declined by increasing protein contents. The highest LDH activity was shown in the isoprothiolane treated fish, while the lowest activity was observed in the isoprothiolane+cartap treatment.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.5
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• pp.1202-1207
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• 2003

In order to clarify the neuroprotective effect of Cornu Saigae Tataricae(CST) water extract on cultured mouse spinal motor neuron damaged by hydrogen peroxide (H₂O₂), MTT [3-(4,5-dimethylthiazole-2-yl)- 2,5-diphenyltetrazolium bromide] assay, LDH (Lactate Dehydrogenase) activity assay and SRB (Sulforhodamine B) assay were carried out after the cultured mouse spinal motor neuron were preincubated with various concentrations of CST water extract for 3 hours prior to exposure of hydrogen peroxide Cell viability of cultured mouse spinal motor neurons exposed to various concentrations of hydrogen peroxide for 6 hours was decreased in a dose-dependent manner. MTT50 values were 40 uM hydrogen peroxide. Cultured mouse spinal motor neurons in the medium containing various concentration of hydrogen peroxide for 6 hours showed increasing of LDH activity and decreasing of total protein synthesis. We know that hydrogen peroxide was toxic on cultured spinal motor neurons. Pretreatment of CST water extract for 3 hours following hydrogen peroxide prevented the hydrogen peroxide-induced neurotoxicity such as increasing of LDH activity and decreasing of total protein synthesis. These results suggest that hydrogen peroxide shows toxic effect on cultured spinal motor neurons and CST water extract is highly effective in protecting the neurotoxicity induced by hydrogen peroxide.

• The Journal of Korea CHUNA Manual Medicine
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• v.2 no.1
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• pp.143-157
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• 2001

Objectives and Methods : To evaluate the mechanism of oxidative damage by xanthine oxydase(XO) and hypoxanthine(HX)-induced oxygen radicals, MTT assay and NR assay were carried out after the cultured mouse spinal sensory neurons were preincubated for 4 hours with various concentrations of XO/HX. And the amount of total protein. neurofilament EIA. lipid peroxidation and LDH activity were measured, to evaluate the protective effect of Herbar Chelidonii(HC) water extract on cultured spinal sensory neurons damaged by XO/HX. after the cultured mouse spinal sensory neurons were preincubated with various concentrations of HC water extract for 3 hours prior to exposure of XO/HX. Results : XO/HX decreased significantly the survival rate of the cultured mouse sensory neurons by NR assay and MTT assay In proportion to concentration and exposed time. In proportion to concentration and exposed time on cultured spinal sensory neurons, XO/HX showed the quantitative decrease of neurofilament by EIA. the decrease of total protein amount by SRB assay and the Increase of lipid peroxidation as well as LDH. HC showed the quantitative increase of neurofilament and total protein, but showed the decrease of lipid peroxidation and LDH activity against the neurotoxicity of XO/HX. Conclusions : From the above results, it is concluded that XO/HX have a neurotoxic effect on cultured spinal sensory neurons and that the herbs extract, such as HC, prevent the toxicity of XO/HX effectively in that they decrease lipid peroxidation and LDH activity.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.27 no.3
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• pp.193-200
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• 2017

Objectives: This study was performed to evaluate the neurototoxicity of the environmental pollutant lead acetate(LA) and the protective effect of the D-2-amino-5-phosphonovaleric acid(APV), N-methyl-D-aspartate(NMDA) receptor antagonist on LA-induced cytotoxicity in cultured C6 glioma cells. Materials and Methods: For this study, cell viability in cultured C6 glioma cells was assessed by XTT assay and antioxidative effect, such as lactate dehydrogenase(LDH) activity, by LDH detection kit. Results: LA significantly decreased cell viability in a dose-dependent manner, and the XTT50 value was determined to be 33.3 uM of LA. The cytotoxicity of LA was deemed highly toxic according to Borenfreund and Puerner's toxic criteria. The vitamin E antioxidant significantly increased cell viability damaged by LA-induced cytotoxicity in these cultures. For the protective effect of APV on LA-induced cytotoxicity, APV significantly increased not only cell viability, but also inhibition of LDH activity. From these results, it is suggested that oxidative stress is involved in the neurotoxicity of LA, and APV effectively protected against LA-induced cytotoxicity via an antioxidative effect as an inhibotory activity of LDH. Conclusions: Natural resources like APV may be putative therapeutic agents for the toxic diminution of environmental pollutants such as LA correlated with oxidative stress.

• Journal of Nutrition and Health
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• v.49 no.2
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• pp.63-71
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• 2016

Purpose: Skin pH, an indicator of skin health, is maintained by various organic factors, which include lactate, free amino acid (FAA), and free fatty acid (FFA). As skin ages or with illness, skin pH becomes less acidic, and functional food has been developed to maintain the acidic pH of skin. In this study, we determined the dietary effect of green tea extract (GTE) on skin pH of photo-aged mice, as measured by epidermal levels of lactate, FAA, and FFA. The protein expression and activity of lactate dehydrogenase (LDH), an enzyme of pyruvate reduction for lactate generation, was further determined. Methods: Albino hairless mice were fed a control diet (group UV+) or a diet with 1% GTE (group GTE) in parallel with UV irradiation for 10 weeks. A normal control group was fed a control diet without UV irradiation for 10 weeks (group UV-). Results: Skin pH was higher (less acidic) in group UV+ than in group UV-. In parallel, epidermal levels of lactate and FFA, as well as of LDH protein expression and activity, were reduced in group UV+. Dietary supplementation of GTE (group GTE) reduced skin pH to similar to the level of group UV-, and inversely increased epidermal levels of lactate, LDH protein expression and activity, but not of FFA. Although epidermal levels of FAA were similar in groups UV- and UV+, it was increased in group GTE to a level higher than in group UV-. In further analysis of major FFA, epidermal levels of palmitic acid [16:0], oleic acid [18:1(n-9)], and linoleic acid [18:2(n-6), but not of stearic acid [18:0] in group GTE were similar to or lower than those in group UV+. Conclusion: Dietary GTE normalized skin pH with increased levels of lactate and FAA, as well as with increased protein expression and activity of LDH in the epidermis of UVB irradiated hairless mice.

• Journal of Veterinary Clinics
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• v.28 no.6
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• pp.549-554
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• 2011

FK506 is a widespread immunosuppressive drug after liver transplantion in patients with advanced-stage hepatocellular carcinoma. Dexamethasone is frequently used as co-treatment in cytotoxic cancer therapy, e.g. to prevent nausea, to protect normal tissue or for other reasons. Our aim was to investigate antitumor effects of FK506 in Hep3B cells, one of differentiated human hepatocellular carcinoma cell lines and inhibitory effects of dexamethsone on FK506- induced antitumor effects. Cell injury was evaluated by biochemical assays as cell viability, lactate dehydrogenase (LDH) and reactive oxygen species (ROS) in Hep3B cells. Intracellular calcium concentration ([$Ca^{2+}$]i) and the level of activation of the c-Jun-N-terminal kinase (JNK) and the Bax protein in cultured Hep3B cells was measured. Exposure of 0.1 ${\mu}M$ FK506 to Hep3B cells led to cell death accompanied by a decrease in cell viability and an increase in LDH, ROS and [$Ca^{2+}$]i. FK506 induced an increase in activity of Bax and JNK protein but inhibited the activity of Bcl-2 protein. Treatment of dexamethsone, per se, had no effects on cell viability, LDH and ROS. However, co-treatment of FK506 and dexamethasone diminished the FK506-induced LDH release, ROS generation and JNK activation. These results demonstrate that FK506 has antitumor effect in Hep3B cells but the combination of FK506 and dexamethasone antagonizes the FK506-induced antitumor effects.

• BMB Reports
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• v.39 no.4
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• pp.426-431
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• 2006

Embryonic stem cells (ESCs), representing a population of undifferentiated pluripotent cells with both self-renewal and multilineage differentiation characteristics, are capable of spontaneous differentiation into cardiomyocytes. The present study sought to define the kinetic characterization of lactate dehydrogenase (LDH) and creatine kinase (CK) of ESC- and neonatal-derived cardiomyocytes. Spontaneously differentiated cardiomyocytes from embryoid bodies (EBs) derived from mouse ESC line (Royan B1) and neonatal cardiomyocytes were dispersed in a buffer solution. Enzymes were extracted by sonication and centrifugation for kinetic evaluation of LDH and CK with spectrophotometric methods. While a comparison between the kinetic properties of the LDH and CK of both groups revealed not only different Michaelis constants and optimum temperatures for LDH but also different Michaelis constants and optimum pH for CK, the pH profile of LDH and optimum temperature of CK were similar. In defining some kinetic properties of cardiac metabolic enzymes of ESC-derived cardiomyocytes, our results are expected to further facilitate the use of ESCs as an experimental model.