• Proceedings of the Korean Society for Emotion and Sensibility Conference
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• 2009.05a
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• pp.97-100
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• 2009

The purpose of this study was to examine effects of treadmill running training on superoxide product of spontaneously hypertensive rats. 20 males aged five week old rats were used for the experiments. They were randomly selected into experimental group (10 rats) and control group (10 rats) after being housed in laboratory for two weeks. The rats of experimental group performed treadmill exercise from 15 minutes for the first time to 35 minutes for the last one in order to familiar with experimental protocol ($VO_{2}max\;65{\sim}70%$, 0.9km/h, 5 times/week, 35 minutes/session). They began to perform treadmill running at the age of eight weeks for eight weeks. Superoxide production and NADPH oxidase activities were measured on the aortal from end of experiment. All statistical analyses and description methods were computed by SPSS Version 10.0. Differences in the values between experimental group and control group were calculated using t-tests. The results of this study were: there wewe no significant differences in superoxide production, NADPH oxidase-drivened superoxide activity, NADPH oxidase activity between two groups (p>.05). Spontaneously hypertensive rats of experimental group were characterized by higher superoxide production and lower NADPH oxidase-drivened superoxide activity and NADPH oxidase activity in comparison with the control group. One possible explanation of this finding was that NADPH oxidase activity of experimental group was lower than the control group. It can be concluded that treadmill running for eight weeks was associated positively with superoxide product of spontaneously hypertensive rats.

• The Korean Journal of Zoology
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• v.27 no.4
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• pp.221-230
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• 1984

The activities of aminopyrine demethylase which is marker enzyme of the microsomal drug-metabolizing system, NADPH-cytochrome a reductase and glutathione peroxidase were measured during the course of liver regeneration after about seventy percent hepatectomy in Wistar rats. In addition, the extent of lipid peroxidation and contents of cytochrome P-450 were also measured. Partial hepatectomy produced a significant depression in aminopyrine demethylase, to reach a minium about 24 hours after operation, but this activity was increased to normal value during regeneration. On the other hand, in sham-operated animals, this showed no change. All the activities of NADPH-chrome P-450 contents of liver microsomes were rapidly decreased at the early stage of regeneration. These values returned to normal after 7 days. By contrast, the activity of glutathione peroxidase was nearly unchanged. According to these results, at the early stage of regeneration, the decrease of cytochrome P-450 and NADPH-cytochrome c reductase activity lead to decrease of lipid peroxidation and drug metabolizing enzyme activity. But these phenomena were not detected after 7 days of regeneration.

• Journal of Nutrition and Health
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• v.23 no.6
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• pp.434-442
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• 1990

A new HPLC method for determining malonaldehyde content in lipid peroxidation systems was developed and antioxidant activities of $\alpha$-, $\beta$- carotene, lutein and lycopene were compared by a newly developed HPLC analysis and by TBA value. In addition, malonaldehyde forming ability of rat liver microsome was determined depending on thawing numbers. As results, malonaldehyde was eluted at a retention o f5.60 min and showed a linear relationship between peak area and concentration in standard curve. The MA content of microsome decreased with thawing numbers possible by destruction of cellular membranes. Lycopene, lutein and $\alpha$-carotene showed stronger antioxidant activities than $\beta$-carotene of DL-$\alpha$-tocopherol both in Fe+3-ADP/NADPH and in paraquat/NADPH system. The inhibitory effects of carotenoids and DL-$\alpha$-tocopherol on Fe+3-ADP/NADPH lipid peroxidation system was similar by TBA value and by the HPLC analysis for malonaldehyde.

• Nutrition Research and Practice
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• v.2 no.2
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• pp.80-84
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• 2008

Beneficial effects of dehydroepiandrosterone (DHEA) supplement on age-associated chronic diseases such as cancer, cardiovascular disease, insulin resistance and diabetes, have been reported. However, its mechanism of action in hepatocellular carcinoma in vivo has not been investigated in detail. We have previously shown that during hepatocellular carcinogenesis, DHEA treatment decreases formation of preneoplastic glutathione S-transferase placental form-positive foci in the liver and has antioxidant effects. Here we aimed to determine the mechanism of actions of DHEA, in comparison to vitamin E, in a chemically-induced hepatocellular carcinoma model in rats. Sprague-Dawley rats were administered with control diet without a carcinogen, diets with 1.5% vitamin E, 0.5% DHEA and both of the compounds with a carcinogen for 6 weeks. The doses were previously reported to have anti-cancer effects in animals without known toxicities. With DHEA treatment, cytosolic malate dehydrogenase activities were significantly increased by ${\sim}5$ fold and glucose 6-phosphate dehydrogenase activities were decreased by ${\sim}25%$ compared to carcinogen treated group. Activities of Se-glutathione peroxidase in the cytotol was decreased siguificantly with DHEA treatment, confirming its antioxidative effect. However, liver microsomal cytochrome P-450 content and NADPH-dependent cytochrome P-450 reductase activities were not altered with DHEA treatment. Vitamin E treatment decreased cytosolic Se-glutathione peroxidase activities in accordance with our previous reports. However, vitamin E did not alter glucose 6-phosphate dehydrogenase or malate dehydrogenase activities. Our results suggest that DHEA may have decreased tumor nodule formation and reduced lipid peroxidation as previously reported, possibly by increasing the production of NADPH, a reducing equivalent for NADPH-dependent antioxidant enzymes. DHEA treatment tended to reduce glucose 6-phosphate dehydrogenase activities, which may have resulted in limited supply for de novo synthesis of DNA via inhibiting the hexose monophophaste pathway. Although both DHEA and vitamin E effectively reduced preneoplastic foci in this model, they seemed to fimction in different mechanisms. In conclusion, DHEA may be used to reduce hepatocellular carcinoma growth by targeting NADPH synthesis, cell proliferation and anti-oxidant enzyme activities during tumor growth.

• Journal of Life Science
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• v.21 no.11
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• pp.1625-1630
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• 2011

Heme oxygenase-1 (HO-1) is a stress-responsive protein that is known to regulate cellular functions such as cell proliferation, inflammation, and apoptosis. In this study, we investigated the role of NADPH oxidase on the expression of HO-1 in human liver hepatoma cell line HepG2. Diphenyleneiodonium (DPI), an NADPH oxidase inhibitor, markedly inhibited HO-1 expression and the nuclear translocation of transcription factor Nrf2 in cobalt protoporphyrin (CoPP) or hemin-treated HepG2 cells. Similarly, the knockdown of $p47^{phox}$, a cytosolic factor for NADPH oxidase activity, by siRNA inhibited the CoPP-induced expression of HO-1. In addition, GSHmee, an intracellular antioxidant, blocked the expression of HO-1 in CoPP-treated cells. Based on these results, we conclude that the blockage of NADPH oxidase with DPI or $p47^{phox}$ siRNA inhibits CoPP-induced HO-1 expression in HepG2 cells, and also suggest that the expression of HO-1 in CoPP-induced HepG2 cells is associated with increase of intracellular ROS by NADPH oxidase activity.

• Applied Biological Chemistry
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• v.36 no.3
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• pp.178-183
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• 1993

Xylose reductase (alditol: $NADP^+$ 1-oxidoreductase, EC 1.1.1.21) from the xylose-fermenting yeast, Candida sp. BT001, was purified via salt fractionation, ion-exchange, gel filtration and affinity chromatography, and its properties were characterized. The enzyme from the yeast was active with both NADPH and NADH as coenzyme. The xylose reductase activity with NADH was approximately 51% of that with NADPH and the specific activities of purified enzyme with NADPH and NADH were 11.78 U/mg and 6.01 U/mg, respectively. Molecular weight of the purified enzyme was 31,000 on SDS-PAGE and 61,000 on gel filtration. The Km for D-xylose, NADPH, and NADH was $94.2{\times}10^{-3}M,\;0.011{\times}10^{-3}M\;and \;0.032{\times}10^{-3}M$, respectively. The purified xylose reductase had relatively higher substrate affinity for L-arabinose than other aldoses tested. The optimal pH was 6.2 and the optimal reaction temperature was $45^{\circ}C$. The thermal stability of the enzyme was for 20 minutes at $30^{\circ}C$.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.778-782
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• 2007

The Diphenyleneiodonium (DPI) is widely used as an inhibitor of flavoenzymes, particularly NADPH oxidase. In this study, we investigated the effect of DPI on the cell growth progression of human colon cancer cells HCT-116 (wild-type p53), HT-29 (p53 mutant) and human breast cancer cells MCF-7 (wild-type p53). DPI treatment in cancer cells evoked a dose- and time-dependent growth inhibition, and also induced the cell cycle arrest in C2/M phase. The peak of cell population arrested in C2/M phase was observed at12 hr after treatment of DPI. In addition, DPI significantly induced the expression of p53, which induces proapoptotic genes in response to DNA damage or irreparable cell cycle arrest, at 6 hr in DPI-stimulated cells. However, a catechol apocynin, which inhibits the assembly of NADPH oxidase, did not induce p53 expression. This suggest that p53 expression induced by DPI is not associated with the inhibition of NADPH oxidase. In conclusion, we suggest that DPI induces the expression of wild-type p53 by ROS-in-dependent mechanism in several cancer cells, and upregulated p53 may be involved in regulatory mechanisms for growth inhibition and cell cycle arrest at C2/M phase in DPI-stimulated cells.

• Journal of Acupuncture Research
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• v.19 no.3
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• pp.95-106
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• 2002

The present study was designed to investigate the effects of various electroacupuncture(EA) stimulation on NADPH-diaphorase and Neuropeptide Y(NPY) in the cerebral cortex of Sprague-Dawley Rats. We evaluated the changes of NADPH-d-positive neurons using a histochemical method and the changes of NPY-positive neurons using a double labeling immunohistochemical method. The staining intensities of NADPH-d-positive neuron and NPY-positive neurons were assessed in a quantitative fashion using a microdensitometrical method based on optical density by means of an image analyzer. As to frequency, the optical density of NADPH-d-positive neurons of 2Hz-1 time EA group was significantly higher than that of 100Hz-1 time EA group in M1, Cg, and that of 100Hz-3 times EA group was significantly higher than that of 2Hz-3 times EA group in Vi, Au, Cg and Ins. As to treatment-time, the optical density of NADPH-d-positive neurons of 2Hz-1 time EA group was higher than 2Hz-3 times EA group in Vi, Au, Cg and Ins. And in Vi 100Hz-1 time EA group and in M1 100Hz-3 times EA group was more efficient. As to frequency, the optical density of NPY-positive neurons of 2Hz-1 time was significantly higher in Vi, and that of 100Hz-1 EA group was higher in M1. And that of 2Hz-3 times EA group was higher than 100Hz-3 times EA group in Cg. As to treatment-time, the optical density of NPY-positive neurons of 2Hz-1 time EA group was significantly higer than 2Hz-3 times EA in S1, Vi, Au. And that of 100Hz-1 time EA group was significantly higher than that of 100Hz-3 time EA in Cg. The present results demonstrated that EA changes the activity in NO and NPY system in the cerebral cortex of SHR and EA stimulation, like frequency and treatment-time, is of importance for this effect.

• Applied Biological Chemistry
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• v.37 no.3
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• pp.203-209
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• 1994

When $^{14}C-{\alpha}-endosulfan$ was incubated with carp liver, kidney and gut preparations, it was metabolized to water soluble and organosoluble metabolites. In an in vitro test, endosulfan was converted to endosulfan ${\alpha}-hydroxyether$ (EHE), endosulfan alcohol (EA) and endosulfan ether (EE). The addition of NADPH resulted in rapid conversion of endosulfan to the metabolites in 105,000 g soluble fraction and microsomes. However, the rate of metabolism of endosulfan in liver, kidney and gut supplemented with NADPH as a cofactor was higher in the 105,000 g soluble fraction than that in the microsomes of carp under incubation conditions. The enzymes probably involved in the metabolism of endosulfan include the glutathione S-transferase (GST) and the mixed function oxidases (MFO), based on the evidence that addition of either GSH or NADPH increased the degradation of endosulfan.

• Journal of environmental and Sanitary engineering
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• v.21 no.1 s.59
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• pp.52-57
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• 2006

Metallothioneins(MTs) belong to the class of low molecular weight proteins. Recently, it has been suggested that MTs may playa direct role in cellular defense against oxidative stress by functioning as antioxidants. Oxidative damage to different cellular components makes a major contribution to many pathogenenesses. Several studies have demonstrated that MT is able to quench a wide range of reactive oxygen species at a higher efficiency than other well known antioxidants such as superoxide dismutate(SOD). The present study was designed to evaluate the effect of MT on the activities of the reactive oxygen species removal system. MT showed the scavenging of superoxide in the SOD assay system in the presence or absence of SOD. When MT was added to nicotinamide adenine dinucleotide phosphate(NADPH) oxidation system in presence of fixed amount of SOD increase the breakdown rate of superoxide. When MT was added to the system that form nitrite from hydroxylammonium chloride, the formation of nitrite was inhibit. We concluded that the function of MT as antioxidant might have an effect on the level of superoxide scavenging.