• Title/Summary/Keyword: cytosolic enzyme

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Cellular Distribution and Metabolism of Ginsenosides in Rat Liver (쥐 간에서의 Ginsenoside의 세포내 분포와 대사)

  • 윤수희;이희봉
    • Journal of Ginseng Research
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    • v.17 no.2
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    • pp.114-122
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    • 1993
  • 0.5 mg of natural ginsenoside mixture and 0.8 $\mu$Ci of synthesized 14C-ginsenosides were administered orally to a rat and killed at one hour after the ginsenoside administration and the liver was fractionated into nuclear fraction, mitrochondria microsomes and cytosol fraction. Radioactivity distribu lion in subcellular fractions of the liver showed that 32o1c of total radioactivity absorbed in the liver was in cytosol fraction but a significant portion of the radioactivity was also found in mitochondria (26.6%) and microsomal fraction (18.l%). 5.8% of the total radioactivity was recovered from the nuclear fraction as well. This suggested that ginsenosides might be distributed into all subcellular fractions. Activities of mitochondrial aldehyde dehydrogenase, lactate dehydrogenase and malate dehydrogenase of the liver of rat at two hours after the ginsenoside administraion were found appreciably stimulated, suggesting that the ginsenoside concentration in the liver might be around 10-5%, since optimum concentrations for most enzyme catalyzed reactions in vitro were known to be 10-6% 10-4%. A significant portion of the radioactivity recovered from subcellular fractions of the liver was found in protein fractions, suggesting that proteins might interact with ginsenosides. Examination of protein-ginsenoside interation by gel filtration, equilibrium dialysis and amonium sulfate precipitation technique suggesting that proteins and ginsenosides do not bound covalently but weakl\ulcorner combined. When purified ginsenoside Rbl and Rgl were incubated with rat liver cytosolic enzymes for 20 min, the above ginsenosides were hydrolyzed quickly, suggesting that ginsenosides might be rapidly hydrolyzed and metabolized in the liver. It was also observed in vitro that the ginsenosides such as Rbl and Rgl were easily hydrolyzed by rat liver cytosol preparation suggesting that absorbed ginsenosides might be quickly hydrolyzed and metabolized in the liver.

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Differentially Expressed Genes by Inhibition of C-terminal Src Kinase by siRNA in Human Vascular Smooth Muscle Cells and Their Association with Blood Pressure

  • Hong, Kyung-Won;Shin, Young-Bin;Kim, Koan-Hoi;Oh, Berm-Seok
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.102-113
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    • 2011
  • C-terminal SRC kinase (CSK) is a ubiquitously expressed, cytosolic enzyme that phosphorylates and inactivates several SRC family protein tyrosine kinases. Recent genomewide association studies have implicated CSK in the regulation of blood pressure. The current study aim is to determine the blood pressure association of the genes regulated by CSK down-regulation. The CSK mRNA expression was downregulated in vascular smooth muscle cells using small interfering RNA (siRNA). CSK mRNA levels fell by 90% in cells that were treated with CSK siRNA; the RNA from these cells was examined by microarray using the Illumina HumanRef-8 v3 platform, which comprises 24,526 reference mRNA probes. On treatment with CSK siRNA, 19 genes were downregulated by more than 2-fold and 13 genes were upregulated by more than 2-fold. Three (CANX, SLC30A7, and HMOX1) of them revealed more than 3 fold differential expression. Interestingly, the HMOX1 SNPs were associated with diastolic blood pressure in the 7551 Koreans using Korea Association REsource data, and the result was supported by the other reports that HMOX1 linked to blood vessel maintenance. Among the remaining 29 differentially expressed genes, seven (SSBP1, CDH2, YWHAE, ME2, PFTK1, G3BP2, and TUFT1) revealed association with both systolic and diastolic blood pressures. The CDH2 gene was linked to blood pressures. Conclusively, we identified 32 differentially expressed genes which were regulated by CSK reduction, and two (HOMX1 and CDH2) of them might influence the blood pressure regulation through CSK pathway.

Effect of Allopurinol Pretreatment on the Liver Damage in $CCl_4$-treated Rat (흰쥐에 있어서 사염화탄소에 의한 간손상에 allopurinol의 영향)

  • 배지혜;윤종국;이상일
    • Toxicological Research
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    • v.11 no.2
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    • pp.247-252
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    • 1995
  • To evaluate the effect of xanthine oxidase on liver injury by $CCl_4$, liver damage was induced both in allopurinol pretreated rats (500 mg/kg. ip) and control group by twice intraperitoneal injection of $CCl_4$ (0.1 ml/100 g body wt. 50% in olive oil) at interval of one day. Increases in the levels of serum alanine aminotransferase and liver weight/body weight (%) by $CCl_4$ were significantly smaller inallopurinol pretreated rats than in control whereas the hepatic microsomal glucose-6-pholphatase activities were significantly higher in allopurinol pretreated rats than control group by $CCl_4$ treatment. These results indicates that allopurinol pretreatment may reduce the liver damage in $CCl_4$ intoxicated rats. In rats either with $CCl_4$or not, hepatic type O xanthine oxidase activities were significantly reduced by allopurinol pretreatment and the increasing rate of these enzymes to each control was remarkably lower in allopurinol pretreated rats than control. Liver cytosolic protein contents and aniline hydroxylase, aminopyrine demethylase activities were higher in allopurinol pretreated rats than coirol rats when animals were treated with $CCl_4$. On the other hand, neither allopurinol pretreated nor $CCl_4$ treatment caused any significant changes in hepatic superoxide dismutase and catalase activities. Hepatic glutathione contents were higher in $CCl_4$-treated rats than control, but no significant changes were found in both between the allopurinol treated rats and $CCl_4$-treated rats pretreated with allopurinol, and glutathione and glutathione S-transferase activities were significantly reduced in $CCl_4$-treated rats than control whereas these enzyme activities showed on significant change in both between allopurinel treated and $CCl_4$-treated rats pretreated with allopurinol. It is concluded that xanthine oxidase reaction system augment $CCl_4$ induced liver injury via even oxygen free radical system.

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Gene Transcription in the Leaves of Rice Undergoing Salt-induced Morphological Changes (Oryza sativa L.)

  • Kim, Dea-Wook;Shibato, Junko;Agrawal, Ganesh Kumar;Fujihara, Shinsuke;Iwahashi, Hitoshi;Kim, Du Hyun;Shim, Ie-Sung;Rakwal, Randeep
    • Molecules and Cells
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    • v.24 no.1
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    • pp.45-59
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    • 2007
  • We describe the gene expression profile of third leaves of rice (cv. Nipponbare) seedlings subjected to salt stress (130 mM NaCl). Transcripts of Mn-SOD, Cu/Zn-SOD, cytosolic and stromal APX, GR and CatB were up-regulated, whereas expression of thylakoid-bound APX and CatA were down-regulated. The levels of the compatible solute proline and of transcripts of its biosynthetic gene, ${\Delta}^1$-pyrroline-5-carboxylate synthetase (P5CS), were strongly increased by salt stress. Interestingly, a potential compatible solute, ${\gamma}$-aminobutyric acid (GABA), was also found to be strongly induced by salt stress along with marked up-regulation of transcripts of GABA-transaminase. A dye-swap rice DNA microarray analysis identified a large number of genes whose expression in third leaves was altered by salt stress. Among 149 genes whose expression was altered at all the times assayed (3, 4 and 6 days) during salt stress, there were 47 annotated novel genes and 76 unknown genes. These results provide new insight into the effect of salt stress on the expression of genes related to antioxidant enzymes, proline and GABA as well as of genes in several functional categories.

Partial Purification and Characterization of Multiple Forms of Extracellular Phospholipase $A_2$ in Human Amniotic Fluid (사람 양수중 다종의 세포외성 포스포리파제의 $A_2$의 부분정제 및 특성)

  • Jeon, Yong-Ju;Baek, Suk-Hwan;Lee, Jee-Hae;Moon, Tae-Chul;Min, Beong-Woo;Chang, Hyeun-Wook
    • YAKHAK HOEJI
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    • v.41 no.2
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    • pp.212-219
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    • 1997
  • Multiple forms of extracellular phospholipase $A_2$ have been detected in human amniotic fluid (HAF). When HAF was subjected to heparin-Sepharose column chromatography, phospholipase $A_2$ activity was detected in both heparin-non binding and binding fraction. The activity of heparin-non binding fraction was further purified by sequential uses of column chromatographies on butyl-Toy-opearl 650M and DEAE-Sephacel. DEAE-Sephacel fraction contained three different phospholipase $A_2$ activities (Peak I, II, III). The molecular weight of DEAE-Sephacel fraction phospholipase $A_2$ determined by SDS-PAGE were about 52KDa (Peak I). Peak II, III required micromolar $Ca^{2+}$ ion for its maximum activity, but Peak I enzyme showed calcium independent phospholipase $A_2$ activity and showed broad range of pH (6.0~10.0) optimum. All these enzymes were not recognized by a monoclonal antibody raised against phospholipase $A_2$ from human synovial fluid. These results suggest that HAF might contain multiple forms of extracellular phospholipase $A_2$, which may neither belong to the 14KDa group II phospholipase $A_2$ family nor cytosolic phospholipase $A_2$.

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Effect of Pine Needle Extract (PNE) on Physiological Activity of SD Rats II. Feeding Effect of PNE on Oxygen Radicals and Their Scavenger Enztmes in Brain Membranes of SD Rats (흰쥐의 생리활성에 미치는 송엽 추출물(PNE)의 영향 II. 뇌세포막의 산소라디칼 및 제거효소의 활성에 미치는 PNE의 투여효과)

  • 최진호;김정화;김동우;김경석;이종수;백영호
    • Journal of Life Science
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    • v.8 no.1
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    • pp.91-96
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    • 1998
  • Pine(Pinus densiflora Sieb et Zucc.) is one of rhe popular plant drugs which has been used as a medicine in Asia. To investigate the effect of pine needle extract (PNE) on oxygen radicals and their scavenger enzymes in brain membranes of Sprague- Dawley (SD), make SD rats were fed basic diets(control group), and experimental diets (PNE group) with 0.5 and 1.0% of PNE 6 weeks. Mitochondrial hydroxyl radical levels in brain of 0.5%-PNE and 1.0%-PNE groups were significantly inhibited to 30% and 25%, respectively, and microsomal hydrogen peroxide levels in brain of 0.5%-PNE and 1.0%-PNE groups were significantly inhibited to 15% compared with control group. Cytosolic superoxide rdical levels in 1.0%-PNE group were significantly inhibited to 20% compared with control group. Lipid peroxide(LPO) levels in brain mitochondria of 0.5%-PNE and 1.0%-PNE groups were significantly lower(25% and 35%) than that in control group. Mn-superoxide disumtase (SOD) activities in brain of 0.5%-PNE and 1.0%-PNE groups were significantly higher(18% and 12%) than those in control groups, but Cu,Zn-SOD activities in brain of 0.5%-PNE were significantly activated to 15% compared with control group. Glutathione peroxidase(GSHPx) activities in brain of 1.5%-PNE and 1.0% PNE groups were significantly higher(14% and 12%) than those in control group. These results suggest that more beneficial effects such as inhibition of oxygen radicals and lipid peroxide(LPO). and oncreases of scavenger enzymes in brain membranes of SD rats may be effectively modulated by administration of pine needle extract (PNE)

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Effects of cranberry powder on biomarkers of oxidative stress and glucose control in db/db mice

  • Kim, Mi Joung;Chung, Jee-Young;Kim, Jung Hee;Kwak, Ho-Kyung
    • Nutrition Research and Practice
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    • v.7 no.6
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    • pp.430-438
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    • 2013
  • Increased oxidative stress in obese diabetes may have causal effects on diabetic complications, including dyslipidemia. Lipopolysccharides (LPS) along with an atherogenic diet have been found to increase oxidative stress and insulin resistance. Cranberry has been recognized as having beneficial effects on diseases related to oxidative stress. Therefore, we employed obese diabetic animals treated with an atherogenic diet and LPS, with the aim of examining the effects of cranberry powder (CP) on diabetic related metabolic conditions, including lipid profiles, serum insulin and glucose, and biomarkers of oxidative stress. Forty C57BL/KsJ-db/db mice were divided into the following five groups: normal diet + saline, atherogenic diet + saline, atherogenic diet + LPS, atherogenic diet + 5% CP + LPS, and atherogenic diet + 10% CP + LPS. Consumption of an atherogenic diet resulted in elevation of serum total cholesterol and atherogenic index (AI) and reduction of high density lipoprotein (HDL)-cholesterol. However, with 10% CP, the increase in mean HDL-cholesterol level was close to that of the group with a normal diet, whereas AI was maintained at a higher level than that of the group with a normal diet. LPS induced elevated serum insulin level was lowered by greater than 60% with CP (P < 0.05), and mean serum glucose level was reduced by approximately 19% with 5% CP (P > 0.05). Mean activity of liver cytosolic glutathione peroxidase was significantly increased by LPS injection, however it was reduced back to the value without LPS when the diet was fortified with 10% CP (P < 0.05). In groups with CP, a reduction in mean levels of serum protein carbonyl tended to occur in a dose dependent manner. Particularly with 10% CP, a reduction of approximately 89% was observed (P > 0.05). Overall results suggest that fortification of the atherogenic diet with CP may have potential health benefits for obese diabetes with high oxidative stress, by modulation of physical conditions, including some biomarkers of oxidative stress.

Effect of SAENGCHINYANGHYOLTANG on the hepatic metabolic enzyme system in streptozotocin-induced diabetic rats (고혈당(高血糖) 쥐의 간(肝) 대사효소계(代謝酵素系)에 미치는 생진양혈탕(生津養血湯)의 영향(影響))

  • Kim, Shin-Seok;Lee, Kyung-Hee;Lee, Cheol-Whan;Choi, Jong-Won;Kim, Seock-Hwan
    • The Journal of Korean Medicine
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    • v.16 no.2 s.30
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    • pp.320-336
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    • 1995
  • SANGNYANGHYOLTANG(SYT) is one of the most important prescription that has been used in oriental medicine for diabetes mellitus. The sudy was done in order to elucidate the anti-diabetic effect of SYT. After pretreatment of SYT(1,000mg/kg) for 6 weeks, the effect of of SYT was prevented on serum liver function test and hepatic lipid peroxide content in rats i.v. injected with streptozotocin(STZ, 50mg/kg, tail vein) 5 weeks after pretreatment of SYT. The hepatic microsomal cytochrome P-450 and aniline hydroxylase were significantly decreased, and aminopyrine N-demethylase activity was significantly increased in SYT-STZ group as compared with control group. Changes in aldehyde oxidase, xanthine oxidase, superoxide dismutase, catalase, epoxide hydrolase, UDP-glucuronyltransferase and sulfotransferase activities were not significantly different in any of the group. The cytosolic glutathione S-transferase activity was significantly decreased in SYT-STZ group as compared with control group. The selenium-independent glutathione peroxidase was significantly increased in SYT-STZ group as compared with control group, but there was no significant difference in selenium-dependent glutathione peroxidase in any of the groups. The hepatic glutathione concentration was significantly increased in SYT-STZ group as compared with control group, and ${\gamma}-glutamylcystein$ synthetase and glutathione reductase activities were not significantly different in any of the groups. The hepatic lipid peroxide content, serum aminotransferase and sorbitol dehydrogenase activities were slightly decreased in significantly in SYT-STZ groups.

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Effects of $\gamma$-Irradiated Beef Feeding on Antioxidative Defense System in Experimental Hepatocarcinogenesis (실험적 간 발암모델에서 감마선 조사 쇠고기 섭취가 쥐의 항산화 방어체계에 미치는 효과)

  • 김정희;진유리;강일준;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.3
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    • pp.646-653
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    • 1999
  • This study was done to investigate the effect of ${\gamma}$ irradiated beef feeding on antioxidant vitamin levels and defense enzyme activities in diethylnitrosamine(DEN) initiated rats. Weaning Sprague Dawley male rats were fed the diet containing ${\gamma}$ irradiated ground beef at the dose 0, 3, 5 kGy as a 20% of protein source for 8 weeks. One week after feeding, rats were intraperitoneally injected twice with a dose of DEN(50mg/kg BW). As a promoter, 0.05% phenobarbital was fed in drinking water from one week after DEN treatment until the end of experiment. At the end of 8th week, serum level of vitamin C, serum and hepatic levels of retinol and tocopherol were determined. In addition, activities of cytosolic glutathione peroxidase, glutathione reductase, glutathione S transferase, catalase and hepatic superoxide dismutase(SOD) were measured. By ${\gamma}$ irradiation, there was no significant effect on serum and hepatic levels of vitamin C and tocopherol except a significant decreasing effect on hepatic retinol level. There was also no significant effect on the activities of enzymes involved in antioxidative defense system, However, DEN treatment led to a significant increase in activities of glutathione reductase and glutathione S transferase while the activity of glutathione peroxidase was decreased. The activities of hepatic SOD and catalase were not changed by DEN treatment. Overall results indicate that the consumption of low dose of ${\gamma}$ irradiated beef does not affect antioxidative defense system.

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Effects of Diallyl Disulfide on the Hepatic Glutathione Peroxidase Activity in Rat (흰쥐 간 Glutathione peroxidase 활성에 미치는 Diallyl disulfide의 영향)

  • Huh, Keun;Lee, Sang-Il;Park, Jong-Min
    • The Korean Journal of Pharmacology
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    • v.22 no.2
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    • pp.144-150
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    • 1986
  • Glutathione peroxidase might play an important role in the protection of cellular structures against oxidative challange by hydrogen peroxide and several organic hydroperoxides. It is widely accepted that allicin is biological active component of garlic, and allicin is easily degraded to diallyl disulfide and other components. This study was attempted to elucidate the effect of diallyl disulfide on some biological activities. It was observed that the activity of serum transaminase and glutathione level in liver were not changed by the treatment of diallyl disulfide. The liver cytosolic glutathione peroxidase activity was significantly enhanced. Whereas, mitochondrial enzyme activity was slightly increased. In the presence of diallyl disulfide in vitro, $V_{max}$ value of glutathione peroxidase for hydrogen peroxide was increased. On the other hand, Km value was not changed.

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