• Title/Summary/Keyword: microsomes

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Comparison of Characteristics of Hepatic Microsomal Cytochrome P45O-dependent Monooxygenases from Snake and Rat (꽃뱀과 흰쥐의 간 마이크로좀에 존재하는 Cytochrome P45O 의존성 Monooxygenases의 특성 비교)

  • Ja Young Moon;Dong Wook Lee;Ki Hyun Park
    • Journal of Life Science
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    • v.8 no.6
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    • pp.695-701
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    • 1998
  • This study was carried out to investigate levels of the components of microsomal mixed function oxidase (MFO) system and activities of the hepatic microsomal cytochrome P45O (P45O)-dependent monooxygenases of grass snake (Natrix tigrina Lateralis) and to compare with those of rat. The levels of P45O and cytochrome b$_{5}$, (b$_{5}$) of snake were much lower than those in rat. NADPH-cytochrome c reductase activity in the snake was also only 40% of that in the rat. Activities of 7-ethoxycoumarin 0-deethylase (ECOD) and benzphetamine N-demethylase (BPDM) of snake hepatic microsomes, when compared with those of rat, were markedly low. But, aryl hydrocarbon hydroxylase (AHH) and testosterone hydroxylase (TSH) activities were nearly the same or higher than those of the rat. Of the P45O-dependent TSHs measured, 7$\alpha$-hydroxylase activity was the highest in snake, whereas, 6$\beta$-hydroxylase activity was the highest in rat. However, stereoselectivity of the enzyme from the snake to C2 and C6 positions of testoste-rone was the same as rat. The result of radioimmunoassay (RIA) for the identification of five P45O isozymes with MAbs shows that relatively high content of ethanol-inducible P45O isozyme, CYP2El, exists in the rat, whereas MC-inducible P45O isozyme, CYP2A1/1A2, does in the snake. From the analyses of SDS-PAGE and RIA of partially pu-rified P45O, we suggest the possibility of the presence of a certain P45O isozyme(s) in hepatic microsomes of snake different from those of rat.

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Effects of Harmaline and Harmalol on the Oxidative Injuries of Hyaluronic Acid, Lipid and Collagen by $Fe^{2+}$ and $H_2O_2$ ($Fe^{2+}$$H_2O_2$에 의한 Hyaluronic Acid, Lipid와 Collagen의 산화성 손상에 나타내는 Harmaline과 Harmalol의 영향)

  • Cho, In-Sung;Shin, Yong-Kyoo;Lee, Chung-Soo
    • The Korean Journal of Pharmacology
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    • v.31 no.3
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    • pp.345-353
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    • 1995
  • ${\beta}-Carboline$ alkaloids including harmaline have been shown to inhibit enzymatically or nonenzymatically induced-lipid peroxidation of microsomes. This study was done to explore the antioxidant ability of harmaline and harmalol on the oxidative injuries of hyaluronic acid, lipid and collagen by $Fe^{2+}$ and $H_2O_2$. Their scavenging actions on reactive oxygen species were also examined. Harmaline, harmalol, superoxide dismutase, catalase and DMSO inhibited both degradation of hyaluronic acid by $Fe^{2+}$ and $H_2O_2$ and lipid peroxidation of microsomes by $Fe^{2+}$. In these reactions, DABCO inhibited degradation of hyaluronic acid but did not affect lipid peroxidation. ${\beta}-Carbolines$ inhibited degradation of cartilage collagen by $Fe^{2+}$, $H_2O_2$ and ascorbic acid. The reduction of ferricytochrome c due to autoxidation of $Fe^{2+}$, which is inhibited by superoxide dismutase, was not affected by harmaline and harmalol. They also did not have a decomposing action on $H_2O_2$. Hydroxyl radical production in the presence of $Fe^{2+}$ and $H_2O_2$ was inhibited by harmaline, harmalol and DMSO. Harmaline and harmalol may inhibit the oxidative injuries of hyaluronic acid, lipid and cartilage collagen by $Fe^{2+}$ and $H_2O_2$ through their scavenging actions on reactive oxygen species, OH and probably iron-oxygen complexes and exert antioxidant abilities.

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The Effect of Excess Dietary Vitamin A on Vitamin K-dependent Carboxylation in Rat Liver Microsomes (비타민 A 과량 섭취가 흰쥐의 간 Microsome의 비타민 K-dependent Carboxylation에 미치는 영향)

  • Lilha Lee
    • Journal of Nutrition and Health
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    • v.25 no.6
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    • pp.492-500
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    • 1992
  • The rate of vitamin K-dependent carboxylation of endogenous liver microsomal proteins and an exogenous peptide substrate for carboxylase were measured to test the effects of excess vitamin A on vitamin K function in rats. In vitro vitamin A incubation in normal rat microsomes of vitamin K-sufficient ras did not influence the carboxylation rates of either endogenous prothrombin precursors or a peptide substrate added, Similarly vitamin A incubation in micro-somes from control and excess vitamin A-fed rats that were on vitamin K-free diet did not change the rate significantly within the respectively groups ; however the rates of endogenous protein carboxylation from excess vitamin A-fed rats tended to be increased by the in vitro vitamin A addition compared to that of control rats. Excess vitamin A-fed rats had 2- to 3- fold higher carboxylase activites of endogenous protein carboxylation either with or without the invitro vitamin A incubation than did control rats. In an in vivo study carboxyalase activites with an added exogenous peptide substrate were not influenced by excess intake of vitamin excess vitamin A-fed rats than for control rats. Carboxylase activites tended to be increased amounts of vitamin A on endogenous protein carboxylation appeareed as early as one week post-initiation of the diet. The results of this study indicate that excess vitamin A produces toxic effect rapidly and that excess dietary vitamin A increase the rate of carboxylation of endogenous protein mainly prothrombin precursors which is an indication of vitamin K defi-ciency.

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The Enhancement of Cytochrome P-450 Mediated Aryl Hydroxylation of Bentazon in Rice Microsomes (벼에서 Bentazon 히드록시화반응에 관련된 Cytochrome P-450 활성(活性) 증진(增進)에 관한 연구(硏究))

  • Pyon, J.Y.;Balke, N.E.
    • Korean Journal of Weed Science
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    • v.17 no.1
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    • pp.59-65
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    • 1997
  • Bentazon 6-hydroxylase (B6H) and cinnamic acid 4-hydroxylase (CA4H) activities were determined in rice (Oryza sativa L.) microsomes to study methods of enhancing cytochrome P-450 mediated aryl hydroxylation of bentazon by hydoxylase inducing compounds. Pretreating rice seeds with 1,8-naphthalic anhydride at 0.5-2% and fenclorim at 5 and 10 ${\mu}M$ increased B6H and CA4H activities. Treatments of rice seedling with ethanol 2.5% enhanced B6H and CA4H activities, and with phenobarbital at 12 mM enhanced B6H activity, and CA4H activity was enhanced at 2 mM. B6H activity was synergistically enhanced by combined treatments of ethanol 2.5 or 5% and phenobarbital 8 or 12mM and also that of 1,8-naphthalic anhydride 0.5 or 1% and phenobarbital 8 or 12 mM, but CA4H activity was decreased by combined treatment. Five-day-old rice seedlings showed higher B6H and CA4H activities which decreased with seedling age.

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Effect of Sam Hwa San Extract on Renal Function in Rabbit (삼화산(三和散)이 가토(家兎) 신장기능(腎臟機能)에 미치는 영향(影響))

  • Jeong, Ji-Cheon
    • The Journal of Dong Guk Oriental Medicine
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    • v.1
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    • pp.55-80
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    • 1992
  • In order to examine that the effect of Sam Hwa San, circulating the vital energy of Sam Cho and controlling body fluid metabolism, gives any influence on renal function, changes in the urine flow, eletrolytes excretion, plasma aldosterone concentration and renin activity were observed after intravenous infusion of the Sam Hwa San extract in rabbit. Also in vitro effect of the herb extract on oxygen consumption in renal cortical slices and ATPase activity in kidney microsomes was measured. The following results were obtained : 1. The urine flow was markedly increased at 10 min after intravenous infusion of the Sam Hwa San extract($0.134{\pm}0.015$ vs. $0.433{\pm}0.046ml/min.kg$), but return ed to normal value after 40 min of infusion. 2. The glomerular filtration rate was significantly increased at 10 min after in travenous infusion of the Sam Hwa San extract, and the renal plasma flow at 10 and 20 min after infusion of the Sam Hwa San extract, following return to normal value. 3. $Na^+$ excretion was significantly increased during 10-40 min after intravenous infusion of the Sam Hwa San extract, although showed the maximal rate at 10-20 min. The fractional $Na^+$ excretion was also increased during 10-40 min. $K^+$ excretion was rapidly increased at 10 min after the intravenous Infusion of the Sam Hwa San extract and then gradually decreased to normal level at 40 min. The fractional $K^+$ excretion was significantly increased during 10-40 min after the intravenous infusion of the Sam Hwa San extract. 4. The plasma aldosterone concentration and renin activity were not altered by the infusion of the Sam Hwa San extract. 5. The ouabain-sensitive oxygen consumption of renal cortical slices was significantly reduced by the Sam Hwa San extract(0.5 and 1.0 vol.%). 6. The Na-K-ATPase activity of renal microsomes was strongly inhibited by the Sam Hwa San extract(0.5 and 1.0 vol.%). These results suggest that the Sam Hwa San causes a strong diuretic effect which results from reduction of Na reabsorption in renal tubule by a direct inhibition of Na-pump and, in part, from all increase in renal blood flow. In clinic, it is considered to obtain the therapeutic effect in body fluid metabolism disharmony to cause the circular disorder of vital energy.

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Effects of Pine Needle Ethyl Acetate Fraction on Membrane Fluidity and Oxidative Stress in Liver Membranes of Rats (간장 세포막의 유동성과 산화적 스트레스에 미치는 솔잎(Pine Needle) 에틸아세테이트획분의 영향)

  • 최진호;김대익;백승진;박시향;김남주;최민경;조원기;김창목
    • 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.

Modulatory Effects of Korean Red Ginseng Extract (Panax ginseng C.A. Meyer) on Cytochrome P450 after Oral Administration to Mice for 14 Days (마우스에 홍삼추출물의 14일간 경구 투여에 따른 약물대사효소 조절능 평가)

  • Kim, Hee-Yeon;Nam, Woong-Shik;Kim, Seong-Hee;Jang, Hye-Ryang;Lee, Mi-Kyoung;Kim, Tae-Wan;Lee, Sang-Kyu
    • Journal of Life Science
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    • v.22 no.8
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    • pp.991-998
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    • 2012
  • Ginseng is one of the most commonly used herbal medicines and health foods. Korean red ginseng (KRG; Panax ginseng C.A. Meyer) extract is known to have potential therapeutic activities, such as anti-viral effects, the amelioration of food allergies, anti-oxidant effects, and obesity reduction. Nevertheless, no reports have been issued the modulatory effects of KRG extract on the activity of cytochrome P450 (CYP). In the present study, we investigated the modulatory effect of KRG extract in vitro and in vivo by using pooled human liver microsomes and male ICR mice. When human liver microsomes were incubated with KRG extract at 0.01-10 mg/ml, CYP1A2, 2B6, 2C19, 2D6, and 3A were not significantly inhibited by KRG extract, although CYP2B6 was slightly inhibited. Mice were orally administered KRG extract at 50, 250, or 500 mg/kg daily for 3, 7, or 14 days. However, the activities of CYPs in mouse livers were not significantly different from those of vehicle-treated controls. In conclusion, no significant ginseng-drug interaction was observed. KRG extract did not significantly modulate the activities of CYPs in vitro or in vivo.

Antioxidative Activity and Produced Condition of Antioxidative Substance by Bacillus sp. FF-7 (Bacillus sp. FF-7에 의한 항산화물질 생산조건과 항산화 활성)

  • Cha, Jae-Young;Kim, Hyo-Jung;Jun, Bang-Sil;Park, Jin-Chul;Ok, Min;Cho, Young-Su
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.165-170
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    • 2003
  • The antioxidative activity of antioxidative substances produced from several bacterial strains isolated from fermented foods were tested by $DPPH\;({\alpha},{\alpha}'-diphenyl-{\beta}-picrylhydrazyl)$ free radical scavenging activity. One of the strains showing the highest antioxidative activity was identified as Bacillus sp. based on the morphological, biochemical, physiological characteristics, and 16S rRNA sequence, and named FF-7. The most optimal medium condition for the production of antioxidative substance from Bacillus sp. FF-7 was 2% galactose as carbon source and l% tryptone as nitrogen source. The antioxidative substance produced from FF-7 in these cultural medium was also tested by in vitro experimental models, the peruxidation of linoleic acid and the peroxidation of rat tissues microsomes by using thiobarbituric acid (TBA) for assay of free malondialdehyde production. The antioxidative activity against lipid peroxidation of rat tissues microsomes was shown in the following order; brain 97.50% > heart 79.95% > kidney 77.84% > spleen 77.47% > testis 69.96% > liver 62.45%. The antioxidative substance produced from FF-7 on linoleic acid peroxidation by IBA method was effectively inhibited during four days, and 0.05% BHT (butylated hydroxytoluene) used comparative control was also effectively inhibited. Results showed that the highest antioxidative activity by DPPH method of antioxidative substance produced from Bacillus sp. FF-7 was obtained by supplementing 2% galactose as carton source and l% tryptone as nitrogen source in cultured medium, this substance effectively inhibited the formation of TBARS in brain microsome in vit개 system and in linoleic acid peroxidation.

In vivo Metabolism of Endosulfan in Carp (Cyprinus carpio) (In vivo 시험에 의한 잉어체내 $^{14}C-endosulfan$의 대사)

  • Lee, K.B.;Shim, J.H.;Suh, Y.T.
    • 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.

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Antioxidative Effects and Chemical Characteristics of Annona muricata Leaf Extracts (그라비올라 잎 추출물의 이화학적 특성 및 항산화 활성)

  • Kim, Young Wan;Kim, Tae Hoon;Ahn, Hee Young;Cho, Young Su
    • Journal of Life Science
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    • v.28 no.5
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    • pp.540-546
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    • 2018
  • Annona muricata, generally known as soursop, graviola, or sirsak, is native to the warmest tropical areas of North and South America and is now widely distributed throughout tropical and subtropical parts of the world, including India and Nigeria. This study tested the contents of polyphenolic compounds, flavonoids, and minerals, as well as the antioxidative effects of DPPH radical-scavenging activity, Fe/Cu-reducing power, linoleic-acid peroxidation using thiobarbituric-acid (TBA) methods and peroxidation of rat-hepatocyte microsomes, and ${\beta}$-carotene bleaching assay. These were tested with in-vitro experimental models using water, ethanol, and methanol extracts of the Annona muricata leaf (AMl). Water extracts of AMl showed the highest extraction yield (1.76%). The total polyphenol-compound concentration was the highest in the methanol extract of AMl. However, the flavonoids concentration was the highest in the ethanol extracts of AMl. AMlMl major minerals were Ca, K, and Mg. In DPPH radical-scavenging activity, the contents exhibited a strong scavenging effect on the ethanol and methanol extracts of AMl. Additionally, the Fe/Cu-reducing power was strong in ethanol and methanol extracts of AMl. $Fe^{2+}$/ascorbate-induced linoleic-acid peroxidation using TBA methods and auto-oxidation of rat-hepatic microsomes showed strong antioxidative activities in ethanol extracts of AMl. ${\beta}$-Carotene bleaching was also highest in the ethanol extracts of AMl. These results may provide the basic data to understand the chemical characteristics and antioxidative effects of Annona muricata leaf extract for the development of functional foods.