• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.1
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• pp.155-159
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• 2002

Effect of the egg yolky from laying hens intubated, p.o., astaxanthin (designated AEY) on the oxidation of mouse liver microtome was investigated using female ICR mouse (6~7 weeks of age). Mice were adapted in a temperature- and humidity-controlled house for one week and randomly divided into 7 treatment groups (10 mice/cage/treatment). Mice were intubated p.o., AEY (50, 100 and 150 mg) or control egg yolk (CEY, 150 mg) every week for 3 weeks. BHT (5 mg) and e -tocopherol (50 mg) were fed to mice as positive control. At week 4, the liver microsome was prepared from sacrificed mice. Protein content of mouse liver microsome with AEY treatment was relatively higher than that with CEY treatment. AEY treatment remarkably lowered the content of unsaturated fatty acids including oleic acid and linoleic acid, but raised that of the saturated fatty acids including stearic acid. AEY group showed relatively higher antioxidative activity than CEY, when used Asc/F $e^{+2}$ or NADPH/F $e^{+2}$ as oxidant. Antioxidative activity of AEY was more effective than $\alpha$-tocopherol, but less effective than BHT.

• The Korean Journal of Pharmacology
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• v.27 no.2
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• pp.155-166
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• 1991

In order to evaluate a potential role of mitochondria in the mediation of toxicity of paraquat (PQ), submitochondrial particle and microsome of mouse liver were compared by oxygen radical generation and lipid peroxidation. With NADH in submitochondrial particle and NADPH in microsome as electron donors, PQ stimulated production of superoxide anion and $H_2O_2$ in both fractions. Under the same conditions, PQ enhanced the generation of ethylene from methional suggestiong stimulation of OH production by PQ. But these effects by PQ were somewhat lower in submitochondrial particle than in microsome. In addition, lipid peroxidation(measured as MDA production) was stimulated by PQ in both fractions. The stimulation of lipid peroxidation in both fractions seemed to occur by the same mechanism probably through perferryl ion. This was supported by the following findings: i) The lipid peroxidation in both fractions was partially inhibited by SOD and completely inhibited by DETAPAC(an iron chelator) but not by catalase or OH scavenger. ii) Addition of $ADP-Fe^{3+}$ further increased PQ-induced lipid peroxidation but decreased ethylene production from methional suggesting no correlation between OH production and lipid peroxidation. The redox-cycling of PQ in mitochondria appeared to be linked to NADH dehydrogenase, not to CoQ since all of the observed stimulations by PQ in submitochondrial particle were inhibited by p-hydroxymercuribenzoate(a NADH dehydrogenase inhibitor) but not affected by other respiratory chain blockers. The above results demonstrate that redox-cycling properties of PQ leading to oxygen radical generation and lipid peroxidation can also occur in mitochondria in the same manner as in microsome. Therefore, the observed actions of PQ in mitochondria suggest that mitochondria may also contribute to toxicity of this drug in vivo.

• Environmental Analysis Health and Toxicology
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• v.10 no.3_4
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• pp.29-40
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• 1995

To investigate and evaluate the scavenging and antioxidative effects of various ftavonoids on paraquat induced pulmonary toxicity, in vivo and vitro tests of eight flavonoids(catechin, epicatechin, flayone, chrysin, apigenin, quercetin, morin and biochanin A) were carried out. In vitro test, inhibitory and antioxidative effects of lipoxygenase dependent lipidperoxidation, NADPH dependent cytochrome p-450 reductase to liver and lung microsome and superoxide anion production in rat peritoneal exudated macrophage were studied. In vivo test, biochemical parameters and cell population in bronchoalveolar lavage fluid(BALF) in mouse and rats after administration of paraquat and flavonoids were tested. The results are summerized as follows; 1. All flavonoids tested inhibited on NADPH dependent cytochrome p-450 reductase in liver and lung microsome. 2. All flavonoids tested showed the inhibitory effects on the superoxide anion production in rat peritoneal exudated macropharge. 3. Lactate dehydrogenase, acid phosphatase and total protein in BALF of mouse which increased by the administration of paraquat, decreased significantly by catechin, chrysin, morin and biochanin A. 4. Numbers of alveolar macropharge and PMN in BALF of rats which increased by the administration of paraquat decreased by all the tested flavonoids. Therefore, all flavonoids tested showed the useful compounds for scavenger and antioxidant on paraquat induced pulmonary toxicity.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.168-173
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• 1995

To investigate the effects of endosulfan on cytochrome P-450 enzymes in mouse(Balb c.), endosulfan was given by an intraperitoneal dose of 7.5 mg/kg. The treatment of endosulfan increased the cytochrome P-450 content by 3.3 to 4.2 fold, cytochrome $b_5$ content by 2.3 to 3.8 fold, NADPH cytochrome P-450 reductase activity by 5.3 to 6.4 fold and total haem content by 3.1 to 3.6 fold of mouse liver after 48 hrs of intraperitoneal injection. Endosulfan cytochrome P-450 absorption spectrum exhibited miximum at 387 nm and 389 nm and broad near 407 nm in the liver microsome. Reduced P-450-CO spectrum of the liver microsome exposed by the treatment of endosulfan showed maximum at 449 nm and 450 nm compared to that of the control having maximum at 451 nm, which indicated endosulfan induced cytochrome P-450 new isozymes. Aldrin epoxidase activities in the mouse liver and kidney were increased by 2.8 and 2.1 fold by the treatment of endosulfan. Also 7-ethoxyresorufin dealkylase activities in the mouse liver and kidney were elevated by 1.7 and 1.8 fold by treatment of endosulfan.

• Applied Biological Chemistry
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• v.38 no.5
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• pp.463-467
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• 1995

In vitro metabolism study of ${\alpha}$-endosulfan by liver and kidney microsomal cytochrome P-450 monooxygenase system of the mouse(Balb/C) was performed. ${\alpha}$-Endosulfan was metabolized to endosulfan lactone(EL), endosulfan hydroxyether(EHE), endosulfan alcohol(EA), endosulfan sulfate(ES), endosulfan ether(EE) and ${\beta}$-endosulfan(${\beta}$-E). The main metabolites of ${\alpha}$-endosulfan were EL(13.2%) and EA(11.5%) in liver microsome and EA(17.4%) md EHE(19.3%) in kidney microsome. The $^{14}C$-activity of organic extractable fraction and water soluble fraction were 63.4% and 31.7% in liver micosome incubates respectively. The water soluble metabolites were EA(83.9%), EHE(4.5%) and ES(2.3). Piperonyl butoxide treatment inhibited the formation of EE by 86%, EA by 92% and EHE, EL and ES were barely formed.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.255-261
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• 2004

Antioxidant activity of extracts from the submerged-liquid culture of mushrooms was measured using two systems : linoleic acid and mouse liver microsomes induced by various free radical sources. Mushrooms of Pleurotus ostreatus (Neutari), Phellinus linteus (Sanghwang), Paecilomyces japonicus (Dongchunghacho), Hericicum erinacium (Norugungdengyee) and Agaricus blazei (Shinryeong) in 1% mulberry tree powder-supplemented medium were incubated in a shaking incubator (200 rpm, $25^{\circ}C$) for 3 days. Hot water extracts of mycelial cultures were freeze-dried, followed by fractioning with hexane, chloroform, ethylacetate and butanol in the order. Antioxidant activity of each sample was examined in free radical-induced linoleic acid oxidation in phosphate-buffered saline (PBS ) solution by measuring the amount of malonaldehyde (MA), and mouse liver microsomal systems by measuring the amount of thiobarbituric acid reactive substances (TBARS). In linoleic acid oxidation system, hot water extracts from the cultures of Pleurotus ostreatus, Phellinus linteus, and Paecilomyces japonicus exhibited stronger antioxidant activity than aqueous or butanol fraction and the combined fraction of hexane, chloroform and ethylacetate, but the hot water extract from Pleurotus ostreatus culture was the strongest activity. The antioxidant activity of the hot water extract from Pleurotus ostreatus culture was stronger than any other fractions in mouse microsomal system. These results suggest that hot water extract of Pleurotus ostreatus culture, and the cultures of Phellinus linteus and Paecilomyces japonicus could be useful for functional materials to reduce the oxidation of lipids in food systems induced by free radicals.

• Toxicological Research
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• v.4 no.1
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• pp.13-22
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• 1988

The inhibitory effect of three polyacetylene compounds, panaxydol, panaxynol and panaxytriol isolated from Panax ginseng C.A. Meyer on $CCl_4$induced lipid peroxidation in vivo and in vitro hepatic microsomal lipid peroxidation induced by ADP-$Fe^{3+}$, NADPH and NADPH-cytochrome P-450 reductase were investigated. Their effects on lowering the lipid peroxide levels both in serum and liver and lowering the serum enzyme (GOT, GPT, LDH) activities without the $CCl_4$-induction were also determined. Male ICR mice were pretreated i.p. with polyacetylene compounds or DL-${\alpha}$-tocopherol before administration of $CCl_4$ i.p. and 20 hr after the administration of $CCl_4,$ serum and liver were analyzed. Hepatic microsome was isolated and used for the in vitro NADPH-dependent lipid peroxidation system. Except for panaxynol, treatment with polyacetylenes to control mice did not reduce the levels of lipid peroxides and serum enzyme activities. Panaxynol itself inhibited lipid peroxidation in the liver of normal mice. Polyacetylene compounds protected from the $CCl_4$-induced hepatic lipid peroxidation and lowered serum lipid peroxide levels. Polyacetylenes also inhibited the in virto hepatic microsomal lipid peroxidation in a dose-dependent manner. The results suggest that panaxydol, panaxynol and panaxytriol seem to be the antioxidant components which contribute the anti-aging activities of Panax ginseng C.A. Meyer.

• The Korean Journal of Food And Nutrition
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• v.15 no.2
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• pp.151-157
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• 2002

This study was carried out to investigate functional activities of the free radical scavenging and germ of Glycin max. Merrill fur cytotoxicity toward P338 and L1210 cells derived from mouse. Effect of crude saponin were examined to oxygen radicals and their scavenger enzymes in liver fractions of Spragae-Dawley(SD) rats. Male rats were fed basic diets of control and experiment diets of 0.5∼1.0% crude saponin. There were no significant differences in hydroxy radical($.$OH) formation of liver mitochondria and microsomes in 1.0% group, while $.$OH formations were significantly decrease in 0.5% and 1.0% saponin compared with control group. Their oxygen radical(O$_2$$\^$$.$/) scavenging activities were significantly decrease in liver cytosol of 0.5% and 1.0% saponin group compared with control group. Soybean germ saponin was isolated purified by the method of HPLC to investigate the cytotoxicity of mouse cells by using the MTT assay. SA-1 saponin fraction of soybean germ showed to inhibit toward growth cell of P338 and L1210 cells and its showed less than 50% cytotoxicity These results suggest that the saponin may play a effective role in attenuating a oxygen radical formations and increasing a scavenger enzyme activities.

• The Korean Journal of Nuclear Medicine
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• v.38 no.4
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• pp.325-329
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• 2004

Purpose: Usefulness of mouse liver S9 fraction was evaluated for the measurement of the metabolites in the in vitro metabolism study of $^{18}F$-labeled radiotracers. Materials and Methods: Mouse liver S9 fraction was isolated at au early step in the course of microsome preparation. The in vitro metabolism studies were tarried out by incubating a mixture containing the radiotracer, S9 fraction and NADPH at $37^{\ciirc}C$, and an aliquot of the mixture was analyzed at the indicated time points by radio-TLC. Metabolic defluorination was further confirmed by the incubation with calcium phosphate, a bone mimic. Results: The radiotracer $[^{18}F]1$ underwent metabolic defluorination within 15 min, which was consistent with the results of the in vivo method and the in vitro method using microsome. Radiotracer $[^{18}F]2$ was metabolized to three metabolites including $4-[^{18}F]fluorobenzoic$ acid within 60 min. It is likely that the one of these metabolites at the origin of radio-TLC was identical with the one that obtained from the in vivo and in vitro (microsome) method. Compared with the in vitro method using microsome, the method using S9 fraction gave a similar pattern of the metabolites but with a different ratio, which can be explained by the presence of cytosol in the S9 fraction. Conclusion: These results suggest that the findings of the in vitro metabolism studies using S9 fraction can reflect the in vivo metabolism of novel radiotracers in the liver. Moreover, this method can be used as a tool to determine metabolic defluorination along with calcium phosphate absorption method.

• The Korean Journal of Pharmacology
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• v.21 no.1
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• pp.1-11
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• 1985

The effects of feeding indole, indole-3-carbinol and benzofuran (all at 5 mmole/kg body wt./day) on various hepatic microsomal and cytosolic enzyme activities involved in xenobiotic metabolism have been compared. Benzofuran was found to elevate the activities of many enzymes both in microsomes (e.g., aniline hydroxylase, 7-ethoxycoumarin O-deethylase, p-nitrophenol UDPGA-transferase and epoxide hydrolase) and in cytosol (e.g., glutathione reductase, glutathione S-transferase, NADH:quinone reductase and UDP-glucose dehydrogenase). The structures of indole and indole-3-carbinol are similar to benzofuran except for the substitution of nitrogen with oxygen atom within the furan ring. Results showed that the activities of UDPGA-transferase and NADH:quinone reductase were not elevated by these indole compounds. While the chemical structure of these two indole compounds are identical except for the presence of the carbinol (methanol) group in indole-3-carbinol, there were marked differences in the types and activities of microsomal enzymes that were enhanced. Among the microsomal enzyme activities determined, indole elevated only the NADPH:cytochrome c reductase, while indole-3-carbinol increased several mixed function oxidase and particularly the epoxide hydrolase activities. Based on the chemical structures of tested compounds and the observed results, possible explanations for the mechanisms involved in elevating epoxide hydrolase activity by benzofuran and indole-3-carbinol are discussed.