• Journal of Life Science
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• v.13 no.5
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• pp.684-691
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• 2003

This study was designed to investigate the effects of ethyl acetate (EtOAc) fraction of pine (Pinus densiflora Sieb et Zucc) needle extract on membrane fluidity (MF), basal and induced oxygen radical (BOR and IOR), lipid peroxide(LPO) and oxidized protein (OP) as an oxidative stress, and lipofuscin(LF) in liver membranes of male Sprague-Dawley rats. Rats were fed basic diets (control) and experimental diets (EtOAc-25, EtOAc-50 and EtOAc-100) for 45days. MFs were significantly increased (about 10%) in mitochondria of EtOAc-100 group compared with control group. BOR and IOR formations in mitochondria were significantly inhibited (about 12∼18% and 9 ∼l2%, respectively) in EtOAc-50 and EtOAc-100 groups, while BOR and IOR formations in microsomes were significantly inhibited (about 9∼l3% and 18∼19%, respectively) compared with control group. LPO levels were significantly inhibited (about 10% and 12∼13%, respectively) in mitochondria of EtOAc-100 and microsomes of EtOAc-50 and EtOAc-100 groups, whereas OP levels were significantly inhibited (about 13∼14%) in mitochondria of EtOAc-50 and EtOAc.-100 groups compared with control group. LF formations were significantly inhibited (about 10∼14%) in these three EtOAc groups. These results suggest that ethyl acetate fraction of pine needle may play an effective role in attenuating an oxidative stress and increasing a membrane fluidity.

• Biomolecules & Therapeutics
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• v.2 no.3
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• pp.236-243
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• 1994

A splenocyte culture system supplemented with liver microsomes was developed to detect immunotoxic chemicals which require metabolic activation using cyclophosphamide as a positive standard. When liver microsomes were added to splenocyte cultures isolated from female B6C3Fl mice, the proliferation of splenocytes by lipopolysaccharide (LPS) was increased and the proliferation by concanavalin A (Con A) was decreased. However, when compared with each corresponding control, cyclophophamide was successfully activated to metabolites capable of suppressing Iymphoproliferative responses. This suppression was clearly dependent upon the amounts of microsomes added and/or the concentration of cyclophosphamide exposed. In these cultures, the proliferation of splenocytes was suppressed when the cells were exposed to cyclophosphamide on the day of culture initiation. On the other hand, microsome was responsible for the increase in LPS mitogenicity and NADPH was responsible for the decrease in Con A mitogenicity. Finally, our present culture system was compared with the hepatocyte-splenocyte coculture system which we had developed earlier. We found that the hepatocyte-splenocyte coculture was better able to activate cyclophosphamide to metabolites capable of suppressing the antibody response to sheep erythrocytes. Although our present culture system was relatively poor to activate cyclophosphamide in cultures for antibody response, it will be useful as a simple screening method to detect suppression of certain in vitro immunotoxic parameters like LPS mitogenicity by chemicals which require metabolism.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.579-585
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• 2003

The studies were carried out on the correlation between microsomal lipid peroxidation level and drug metabolizing enzyme activities in rat liver microsomal suspensions on various ages (2-week-old, 2, 4, 8, and 12-month-old). The lipid peroxidation levels of liver homogenates tended to be elevated in a 4-month-old rat livers, but it was a little decreased in 8 and 12-month-old rat livers. The lipid peroxidation levels of microsomal suspension was not shown any significant differences by ages. Lipid peroxidation levels and microsomal cytochrome P450 and NADPH-cytochrome c reductase activity showed a direct correlation (r=0.72 and r=0.64), respectively. The activities of cytochrome P450-dependent aminopyrine-N-demethylase and benzpyrene hydroxylase in rat liver microsomes were increased by ages up to 8-month-old rats and maintained in 12-month-old rats. The correlation between lipid peroxidation levels and these cytochrome-dependent enzyme activities showed a high direct correlation (r=0.97 and r=0.81), respectively.

• YAKHAK HOEJI
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• v.43 no.6
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• pp.834-842
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• 1999

The metabolism and pharmacokinetics of M-l, which is metabolite of pentoxifylline, have been studied in human liver microsomes. Biphasic kinetics was observed from the Eadie-Hofstee plot for the formation of both metabolites of M-l. For the kinetics of pentoxifylline, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 1,648 and 5,622 nmol/min/mg protein, and the estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.180 and 4.829 mM, respectively. For M-3, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 0.062 and 0.491 nmol/min/mg protein, and estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.025 and 1.216 mM. The formations of pentoxifylline and M-3 from M-1 were indentified by using several selective inhibitors of cytochrome P450 isoformes at 0.05-5 mM concentration of M-1 in human liver microsomes. For the analysis of low (0.05 mM) concentration of M-1, where the affinity was expected as low, indicated that CYPlA2 and CYP3A4 were major P450 isoforms responsible for pentoxifylline and M-3 formation. CYP3A4 and CYP2A6 appeared to be P450 isoforms responsible for M-3 formation at high (5 mM) concentration of M-1.

• The Korean Journal of Community Living Science
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• v.20 no.2
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• pp.223-230
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• 2009

This study is to find out the antioxidative effect and serum and liver lipid composition of wild grape juice in vivo. Forty 6-week-old white Sprague Dawley rats were divided into 4 groups such as normal lipid group, normal lipid group with wild grape juice, oxidized lipid (basic diet plus 10% of oxidative lipid) group and oxidized lipid group with wild grape juice, and 2ml juice was provided everyday. After 4 weeks of feeding with experimental diet each groups were examined for the antioxidant enzyme activity in blood and liver microsome and their serum and liver lipid composition. Glutanthione peroxidase activity in blood was significantly higher in oxidized lipid group with wild grape juice than in oxidized lipid group. Glutanthione peroxidase activity showed no difference depending on wild grape juice supplementation. Glutanthione peroxidase activity in liver was significantly higher in the groups with wild grape juice than in the groups supplemented only with oxidized lipid. Glutanthione reductase activity showed no difference depending on the supplementation of wild grape juice. Serum triglyceride level in the group supplemented with oxidized lipid diet and wild grape juice showed similar value to the normal lipid group and the normal lipid group with wild grape juiceoxidized fa6. Liver total lipid in the group supplemented with oxidized lipid and wild grape juice showed similar value to the normal lipid group and the group supplemented with normal lipid and wild grape juice. And it was lower than that of oxidative lipid group without juice. The liver triglyceride level in the group supplemented with normal lipid and wild grape juice was lower than that in the oxidative lipid group, but it was as low as in the group supplemented only with normal lipid.

• Korean Journal of Plant Resources
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• v.13 no.3
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• pp.171-178
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• 2000

This study was performed to investigate the possible effects of some plants protecting intact rat liver damaged by $CCl_4$. The extract of mugwort (Artemsiae capillaris), soybean sprout and pine leaf (Pinus strobus) inhibited markedly the in vitro activities of rat liver fatty acid synthase, whereas those of shiitake (Lentinus ododes), Houttuynia cortata, Acanthopanacis cortex and buckwheat leaves had less effects. Treatment with the water extract of pine leaf and soybean sprout caused a marked decrease in the $CCl_4$-induced toxicity in rat liver, judged from their effects on the levels of glutamic oxaloacetic transaminase (GOT) and glutamate pyruvic transaminase(GPT) in the serum. The extract of mugwort and soybean sprout reduced markedly the content of liver microsomal peroxides induced by $CCl_4$ treatment and serum TBA values, respectively. The extract of soybean sprout decreased efficiently the content of liver triglyceride elevated by $CCl_4$ treatment. Nevertheless, the extracts did not exert the supression of hepaticmegaly induced by $CCl_4$. The results suggest that soybean sprout and pine leaf may be potential sources improved the biochemical parameters like as peroxidation value or serum GOT and GPT, although these extracts had minimal effects in the increase of liver size induced by carbon tetrachloride.

• Journal of Society of Preventive Korean Medicine
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• v.7 no.2
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• pp.65-74
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• 2003

Objective : The aim of present study is to evaluate the inhibitory potential of licorice extract and glycyrrhizin on cytochrome P450(CYP) in human liver microsomes. Methods : Using human liver microsomes, water extract of licorice and glycyrrhizin as an inhibitor were co-incubated with each probe drug representing selective CYP isoform activity. We measured relative metabolic activity in incubation condition compared to that with no extract of licorice using HPLC system. Results : Both water extracts of licorice and glycyrrhizin showed inhibitory effect on CYP-catalyzed reactions. CYP2C19 $(IC_{50}=126.7{\mu}g/ml)$ is most potently inhibited by water extract than other tested CYP isoforms$(IC_{50}>450{\mu}g/ml)$, but glycyrrhizin exhibited potent inhibition on CYP1A2$(IC_{50}=106.9{\mu}g/ml)$ followed by CYP2C9 and CYP2D6. Conclusion: These results indicate that water extract of licorice and glycyrrhizin have inhibitory potential on CYP-catalyzed reaction in human liver microsomes. But the mechanism of inhibition was slightly different between them Water extract of licorice mainly inhibited CYP2C19, and glycyrrhizin primarily inhibited CYP1A2. The inhibition by water extract of licorice and glycyrrhizin on CYP isoforms may cause drug interaction with co-administered drug leading to toxicity or treatment failure.

• Journal of Life Science
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• v.12 no.2
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• pp.136-143
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• 2002

Jaboyangyeong-hwan (IAE) has been known as a traditional medicine for the treatment of debility, fatigue, and liver diseases. The hepatoprotective effect of the water extract of Jaboyangyeong-hwan was investigated against carbon tetrachloride ($CCl_4$)-induced hepatic damage. A single intraperitoneal injection of $CCl_4$produced liver damage in rats as manifested by the significant rise of aspartate aminotransferase (AST, alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in serum as compared to those of untreated normal group. Pretreatments of rats with the JAE extract (300, 600, and 1200 mg/kg for 7 days) were significantly reduced AST, ALT, and ALP levels compared with $CCl_4$-treated control group. Treatment of rats with $CCl_4$led to significantly increase in lipid peroxidation and significantly decrease in cytochrome P450 and P450 reductase. The oral administration of the JAE extract significantly inhibited the accumulation of microsomal thiobarbituric acid reactive substance (TBARS) and increased the cytochrome P450 and P450 reductase activity. All these biochemical alterations resulting from $CCl_4$administration were inhibited by the pretreatment with JAE extract. These results suggest that JAE water extract can be useful as a hepatoprotective agent.

• Environmental Analysis Health and Toxicology
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• v.10 no.1_2
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• pp.47-54
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• 1995

To investigate and evaluated the scavenging and antioxidative effects of various flavonoids on paraquat induced toxicity, in vivo and vitro tests of eight flavonoids (catechin, epocatechin, flavone, chrysin, apigenin, quercetin, morin and biochanin A) were carried out. The generation of reactive oxygen substances(ROS) in PMS-NADH system $H_2O_2$ induced hemolysis and lipidperoxidation to blood, NADPH dependent lipidperoxidation to liver and lung microsome by paraquat were studied.The results are summerized as follows; 1) In the concentration ranges from 3.3 to 9.8$\mu$M of catechin,epicatechin, quercetin and biochanin A removed the 50% of DPPH radical scavenging effects. 2) In the concentration ranges from 0.60 to 1.86 mM of catechin, epicatechin, quercetin and biochanin A showed the inhibitory and antioxidative activity on superoxide anion which gernerated in PMA-NADH system. 3) In the concentration ranges from 0.12 to 0.49mM of catechin, epicatechin, quercetin and biochanin A showed the inhibitory and antioxidative activity on H202 which generated in PMA-NADH system. 4) In the concentration ranges from 0.6 x10$^{-5}$ to 6.3 x 10$^{-5}$mM of catechin, epicatechin, flavone, chrysin, quercetin and morin showed the inhibitory and antioxidative activity on $H_2O_2$ induced hemolysis to blood 5) All flavonoids tested exhibited inhibitory and antioxidative effects on paraquat induced liver and tung microsomal lipidperoxidation. Therefore, all flavonoids evaluated showed the useful compounds for scavenger and antioxidant on paraquat induced toxicity.

• YAKHAK HOEJI
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• v.28 no.1
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• pp.53-59
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• 1984

Drug-metabolizing system which has the important role in drug metabolism is localized in smooth endoplasmic reticulum of hepatocytes and is composed of NADPH, NADPH-cytochrome $P_{450}$ reductase, cytochrome $P_{450}$ and others. It is well known that the enzyme system is induced by phenobarbital and methylcholanthrene. Lipid peroxidation is reaction of oxidative deterioration of polyunsaturated lipids. Formation of lipid peroxides in liver microsome has been found to produce degradation of phospholipid, which are major components of microsomal membrane. The relationship between the formation of lipid oxides and the activities of drug-metabolizing enzyme in the liver of rats was reported by several investigators. In this study the effect of riboflavin tetrabutylate, an antioxidant on lipid peroxidation, specially the relationship between lipid peroxidation and drug-metabolizing enzyme system was investigated. In addition the effect of vitamin A derivatives, such as retinoic acid and retinoid on the enzyme was also observed. Results are summarized as followings. 1) The pretretment with riboflavin tetrabutylate inhibited completely the lengthened sleeping time due to $CCl_{4}$ treatment. 2) The increase of TBA value was prevented by the pretreatment with riboflavin tetrabutylate. 3) The pretreatment with riboflavin tetrabutylate also prevented the decrease of drug-metabolizing enzyme caused by $CCl_{4}$. 4) Both retinoic acid and retinoid remarkably decreased the activity of aminopyrine demethylase. Pretreatment of riboflavin tetrabutylate, however, prevented inhibitory effect of retinoic acid on the enzyme activity.