• Title/Summary/Keyword: Rat liver homogenate

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Effect of G009 on $CCl_4-Induced$ Hepatic Injury and Lipid Peroxidation in Rats (G009가 $CCl_4$로 유발된 간손상 및 지질 과산화에 미치는 영향)

  • Jeong, Hoon;Han, Man-Deuk;Baek, Sung-Jin;Kim, Yong-Seok;Kang, Sang-Mo;Lee, June-Woo
    • Korean Journal of Pharmacognosy
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    • v.27 no.3
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    • pp.159-166
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    • 1996
  • To evaluate hepatoprotective effects of G009, an hepatoprotective agent which was extracted from the mycelia of Ganoderma lucidum IY009, we were, studied using $CCl_4$-and galactosamine-induced hepatotoxicity in rats. The ratio of liver weight to body weight, the value of glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase (GPT) activities, the change of a lipids in serum, and the inhibitory activity of malondialdehyde (MDA) formation in serum and liver homogenate were determined in rats. G009 was not significantly changed of the ratio of liver weight to body weight and the content of lipids in serum, but reduced the serum GOT and GPT values in $CCl_4$-and galactosamine-induced hepatotoxicity in rat. Especially, protective effect of G009 on rat hepatic injuries induced by galactosamine was significantly appeared. $CCl_4$ increased markedly the formation of lipid peroxides in the liver homogenate, and serum. The increase of lipid peroxides by $CCl_4$-induced hepatotoxicity was markedly reduced by the treatment with G009. These results suggest that the hepatoprotective effects of G009 may be correlated with its anti-lipid peroxidative activity, therefore, it may be potential agent for hepatic disease.

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Effects of Rhynchosia molubilis Saponin on Hepatotoxicity and Pathology (간독성과 병리학적인 면에서 노두 사포닌의 효과)

  • Ha Bae-Jin
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.186-191
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    • 2006
  • This study was carried out to investigate the effects of Rhynchosia molubilis saponin on carbon tetrachloride ($CCl_4$)-induced hepatotoxicity. Sprague-Dawley rats were intraperitoneally administered the Rhynchosia molubilis saponin at 100 mg/kg every day for two weeks, then $CCl_4$ (3.3 ml/kg) was injected into rats. 12 hours later, they were anesthesized with ether and dissected. Rhynchosia molubilis saponin-administered group showed 59.92% and 62.28% of inhibitory effects on aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities compared to $CCl_4$-treated group (p<0.05). Malonedialdehyde (MDA) levels of Rhynchosia molubilis Saponin-administered and $CCl_4$-treated (RSC) group in liver homogenate and mitochondria were significantly inhibited to 61.83%, 81.11 %, respectively, compared to $CCl_4$-treated group (p<0.05). Superoxide dismutase (SOD) activities of RSC group in liver homogenate and mitochondria were significantly inhibited to 66.53%, 31.04%, respectively, compared to $CCl_4$-treated group (p<0.05). GPx activities of RSC group in liver homogenate and mitochondria were significantly inhibited to 72.74%, 72.68%, respectively, compared to $CCl_4$-treated group (p<0.05). The histological examinations showed that the liver cell necrosis and centrilobular congestion aggregation induced by $CCl_4$ were dearly eliminated by the administration of Rhynchosia molubilis saponin. These results suggest that Rhynchosia molubilis saponin could have the protective effects against hepatotoxicity.

Rat Liver 10-formyltetrahydrofolate Dehydrogenase, Carbamoyl Phosphate Synthetase 1 and Betaine Homocysteine S-methytransferase were Co-purified on Kunitz-type Soybean Trypsin Inhibitor-coupled Sepharose CL-4B

  • Kim, Hyun-Sic;Kim, Ji-Man;Roh, Kyung-Baeg;Lee, Hyeon-Hwa;Kim, Su-Jin;Shin, Young-Hee;Lee, Bok-Luel
    • BMB Reports
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    • v.40 no.4
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    • pp.604-609
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    • 2007
  • An Asp/His catalytic site of 10-formyltetrahydrofolate dehydrogenase (FDH) was suggested to have a similar catalytic topology with the Asp/His catalytic site of serine proteases. Many studies supported the hypothesis that serine protease inhibitors can bind and modulate the activity of serine proteases by binding to the catalytic site of serine proteases. To explore the possibility that soybean trypsin inhibitor (SBTI) can recognize catalytic sites of FDH and can make a stable complex, we carried out an SBTI-affinity column by using rat liver homogenate. Surprisingly, the Rat FDH molecule with two typical liver proteins, carbamoyl-phosphate synthetase 1 (CPS1) and betaine homocysteine S-methyltransferase (BHMT) were co-purified to homogeneity on SBTI-coupled Sepharose and Sephacryl S-200 followed by Superdex 200 FPLC columns. These three liver-specific proteins make a protein complex with 300 kDa molecular mass on the gel-filtration column chromatography in vitro. Immuno-precipitation experiments by using anti-FDH and anti-SBTI antibodies also supported the fact that FDH binds to SBTI in vitro and in vivo. These results demonstrate that the catalytic site of rat FDH has a similar structure with those of serine proteases. Also, the SBTI-affinity column will be useful for the purification of rat liver proteins such as FDH, CPS1 and BHMT.

Effect of p-Phenylenediamine Application to Rat Skin on the Liver Oxygen Free Radical Systems

  • Park, Hye-Jung;Lee, Sang-Hee;Yoon, Chong-Guk
    • Biomedical Science Letters
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    • v.9 no.2
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    • pp.75-84
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    • 2003
  • To evaluate the influence of hepatic oxygen free radical systems on liver injury by topical p-phenylenediamine (PPD) application on rat skin, PPD (25 mg/16.5 $\textrm{cm}^2$) was topically applied to the abdominal region 5 times every other day and sacrificed. By PPD treatment, increasing rate of liver weight/body weight (%), serum activities of alanine aminotransferase and aspartate aminotransferase and decreasing rate of microsomal glucose-6-phosphatase activity were higher in the rats fed tungstate supplemented diet than those fed a standard diet. These findings indicate that group fed tungstate supplemented diet have more severe liver injury compared with group fed standard diet on topical PPD application. However, the activities of oxygen free radical generating enzymes such as xanthine oxidase (XO) and cytochrome P450 dependent aniline hydroxylase and those of oxygen free radical scavenging enzymes were not found to be different between these two animal groups. In the present study, a novel monitoring method to detect the generating of oxygen free radicals in liver extract was devised. Throughout this method, the oxidized PPD produced by oxygen free radicals was determined colorimetrically. The increasing rate of PPD oxidation by liver homogenate was higher in tungstate fed animals than in standard diet fed ones. Among the fractionations of liver extract, the mitochondrial and postmitochondrial fractions in the liver extract of tungstate fed animals led to a higher availability of PPD oxidation by PPD treatment compared with standard diet fed ones. In conclusion, these results suggest that an enhanced liver injury in tungstate fed animals treated with PPD may be due to oxygen free radicals produced in other systems except oxygen free radicals generating from cytosolic XO system. Especially, oxidative availability by PPD can be used for oxygen free radical detection in some tissue.

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Study on the Effect of Concentrated Extracts of Pteridium sp. Tube Feeding on the Serum Lipid Peroxides and Hepatic Homogenate Lipid Peroxides value (Pteridium sp.의 농축추출액(濃縮抽出液)이 동물혈청(動物血淸) 및 간조직(肝組織)의 지질과산화(脂質過酸化)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Lee, Yong-Ock
    • Journal of the Korean Applied Science and Technology
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    • v.3 no.2
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    • pp.1-8
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    • 1986
  • This experiment was carried out to investigate variation of plasma lipid level and liver peroxide level(Thiobarbituric acid, T.B.A) caused by tube feeding of Pteridium sp. extracts for seven weeks in rats. A seven week rat's plasma triacylglyceride (T.G.) level showed remarkably higher than that of the control group and that of the three week rats. On the other hand, no remarkable differences of plasma phospholipid, free fatty acid and thiobarbituric acid level have been recognized. Liver homogenate lipid peroxides value of seven week feeding rats showed remarkably higher level compared with that of the control group and the three weeks rats. So it could not be agreed that short time feeding of Pteridium sp. plant or even it's concentrated extracts influenced to the levels of liver homogenate lipid peroxides, plasma lipids and its peroxides.

Antioxidative Activity of Zinc-Enriched Saccharomyces cerevisiae FF-10 in In vitro Model Systems (아연-고함유 효모 Saccharomyces cerevisiae FF-10 세포액의 항산화효과)

  • Cha, Jae-Young;Park, Bo-Kyung;Ahn, Hee-Young;Eom, Kyung-Eun;Jun, Bang-Sil;Cho, Young-Su
    • Journal of Life Science
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    • v.19 no.2
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    • pp.179-184
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    • 2009
  • Zinc is an essential trace element for human and plays an important biological role in antioxidant properties. We have been reported that zinc-enriched S. cerevisiae FF-10 contained 392 mg% in the YM basal and 3,193 mg% in the YM optimal medium. Antioxidative activity of FF-10 was tested in vitro models by DPPH (${\alpha},{\alpha}'$-diphenyl-${\beta}$-picrylhydrazyl) radical scavenging activity and lipid peroxidation using linoleic acid (LA) and rat liver homogenate. DPPH radical scavenging activity was higher in the cell-free extract of FF-10 cultured in the YM optimal medium (YMOM) than that in the YM basal medium (YMBM). The inhibition activity of lipid peroxidation using rat liver homogenate was shown in the following order: BHT > YMOM > YMBM and these values were dose dependently. The lipid peroxidation of the control mixture by ferric thiocyanate and TBA methods using LA was increased rapidly as typical peroxidation curve of LA from one day and the antioxidation activity of the cell free extracts by cultivating FF-10 in the YMOM were higher than that of the YMBM. Result of this study indicate that the cell-free extracts containing a high intercellular zinc of S. cerevisiae FF-10 cultured in YMOM showed strong antioxidation capacities in DPPH radical scavenging activity and lipid peroxidation using LA and rat liver homogenate.

The Anti-Oxidative Effects of Oldenlandiae Diffusae Herba Extract on Oxidative Hepatic Injury (간세포(肝細胞)의 산화적 손상에 대한 백화사설초(白花蛇舌草)의 항산화효과)

  • Kim, Hyeong-Hwan;Lee, Chae-Jung;Park, Chul-Soo;Kim, Mi-Rang;Kim, Jong-Dae;Mun, Jin-Young;An, Jung-Hwan
    • The Journal of Internal Korean Medicine
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    • v.23 no.1
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    • pp.57-64
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    • 2002
  • Objective : This study was designed to investigate the anti-oxidative effects of Oldenlandiae Diffusae Herba Water extract (ODHW) on lipid peroxidation by free radicals oxidative hepatic injury. Methods : In order to evaluate anti-oxidative activities of ODHW in the liver cell, cultured normal rat liver cells(Ac2F) were incubated with or without ODHW. After 16 hours to 18 hours of experiment, cells were placed in DMEM medium without serum, and then incubated with 1mM tert-butyl hydro-peroxide(t-BHP) for two hours. Viable cells were detected by MTT assay. The levels of LPO induced by hydroxyl radical derived from H2O2-Fe2+ system in rat liver homogenate were determined by means of TBA. Inhibitory effect of ODHW on superoxide generation was measured by xanthine-xanthine oxidase system. Results : In the linoleic acid autoxidation system, ODHW exhibited antioxidant activity, which inhibited 85% of linoleic acid peroxidation. These effects were similar to those of dl-a-tocopherol. ODHW showed scavenging effects on DPPH radical, inhibited superoxide generation in xanthine-xanthine oxidase system, and also inhibited lipid peroxidation of rat liver tissue with hydroxyl radical derived from $H_2O_2-Fe^{2+}$ system. In addition, ODHW protected the cell death induced by t-BHP and it significantly increased cell viability in a normal rat liver cell(Ac2F)

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Effects of Amino Acids on the Activities of Alcohol Metabolizing Enzyme Alcohol Dehydrogenase (ADH) and Acetaldehyde Dehydrogenase (ALDH) (알코올 대사 효소 alcohol dehydrogenase (ADH) 및 acetaldehyde dehydrogenase (ALDH) 활성에 미치는 아미노산의 영향)

  • Cha, Jae-Young;Jung, Hae-Jung;Jeong, Jae-Jun;Yang, Hyun-Ju;Kim, Yong-Taek;Lee, Yong-Soo
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1321-1327
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    • 2009
  • The present study examined the comparative effects of various amino acids on the alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities of yeast Saccharomyces cereviciae and rat liver homogenate in vitro. Methionine showed the highest activity in yeast ADH among the amino acids used in this study, but this was not higher than that of the hangover product, Condition-Power (CP) used as positive control. Methionine was also found to be the best amino acid in terms of the ALDH activity in rat liver homogenate among the treatment amino acids, which was comparatively higher than that of positive control CP. It was chosen for further experiments and yeast ADH activity increased in parallel with increased methionine concentration, but not rat liver ALDH activity, and it was comparatively higher than those of the positive control. Arginine showed the highest values in yeast ALDH and rat liver ADH activities among amino acids, and it was chosen for further experiments. Yeast ALDH activity increased in parallel with increased arginine concentration, which was higher than that of positive control CP, and rat liver ADH activity was also comparatively higher in all treatment concentrations of arginine than that of positive control CP. The native electrophoresis of ADH and ALDH from cell-free extracts of yeast Saccharomyces cerevisiae cultured in the growth medium containing various arginine concentrations by $0{\sim}0.1%$ showed two active bands upon zymogram staining analysis, and the straining intensity of ADH and ALDH active bands in arginine treatment yeast was stronger than that of non-yeast or low treatment yeast. These results indicate that alcohol metabolizing enzyme activities can be enhanced by arginine and methionine, suggesting that arginine and methionine have potent ethanol-metabolizing activities.

Transferrin Receptors in the Liver Cell Membrane of Carcinogen (3-methyl-4-dimethyl-arninoazobenzene) Treated Rat (Carcinogen (3-methyl-4-dimethyl-aminoazo benzene) 처리후 간세포막에서의 Transferrin Receptor 변동에 관한 연구)

  • Lee, Jae-Heun;Ro, Eu-Sun;Hur, Gang-Min;Lee, Choong-Sik;Seok, Jeong-Ho
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.85-96
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    • 1993
  • To investigate the alteration of transferrin receptor (TfR) in the proliferating or transformed liver cells, $^{125}I-transferrin$ binding experiment was carried out in the isolated parenchymal cells (PC) or nonparenchymal cells (NPC) from normal regenerated rat liver after partial hepatectomy and from the liver of 3-methyl-4-dimethyl-aminoazobenzene (3-Me-DAB) treated rat. With the administration of 3-Me-DAB for 8 weeks, the liver tissue showed marked morphologic changes of oval cell proliferation, regenerations of hepatocytes, and atypical proliferations of bile ducts, but these changes were little affected by partial hepatectomy. Transferrin binding values in PC or NPC homogenate from the regenerated liver of normal rat, were increased by 3rd day and diminished to control level at 7th day after partial hepatectomy. With the treatement of 3-Me-DAB for 8 weeks, transferrin binding sites in homogenates were higher than those of normal rat liver and increased by 7th day after partial hepatectomy. Transferrin binding sites (Bmax) in the cell membrane of NPC were higher than those of PC of normal rat liver, but there was no significant difference in Kd values between both groups (5.05, 6.3 nM). In the normal resenerated rat liver, transferrin binding sites in the PC or NPC plasma membrane, were increased by 3rd day and diminished to control level at 7th day after partial hepatectomy. With 3-Me-DAB tratment, transferrin binding sites in both liver NPC and PC plasma membrane were increased about 3 folds, compared to those in each plasma membrane of normal rat liver. And after partial hepatectomy of 3-Me-DAB trated rat, transferrin binding sites were increased by the 3rd day in the NPC plasma membrane but increased by the 7th day in the PC plasma membrane. In the transferrin binding sites of the PC or NPC plasma membrane of 3-Me-DAB treated liver, two kinds of Kd values $(3.1{\sim}4.7\;nM,\;25.4{\sim}54.1\;nM)$ were detected. The present results suggest that 1) TfRs are distributed in the liver PC as well as NPC; 2) Increased TfRs in PC or NPC plasma membrane of normal regenerated liver after partial hepatectomy and 3-Me-DAB treated rat liver, may be due to increased intracellular synthesis; 3) Increased TfRs in normal regenerated liver after partial hepatectomy might be related to the expression of a single type of high affinity site $(Kd,\;3.1{\sim}7.5\;nM)$, but in 3-Me-DAB treated rat liver might be related to the expression of high and low affinity types of receptors $(Kd,\;25.4{\sim}54.1\;nm)$.

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Effects of Isatis indigitica on Anti-lipid Peroxidation and Low Density Lipoprotein Oxidation (판람근의 지질과산화 및 저밀도지단백산화에 미치는 영향)

  • 장현진;양기숙
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.448-451
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    • 2002
  • Isatis indigotica is commonly used as traditional chinese medicine for antipyretic, antiviral, and detoxifying purpose in China. In order to examine whether it prevents lipid peroxidation, root of Isatis indigotica was extracted with methanol and fractionated with hexane, ethyl acetate, butanol and water. The activity was evaluated by DPPH method and lipid peroxidation in rat liver homogenate and C $u^{++}$ -induced oxidated LDL. The results showed that ethyl acetate fraction and indigo, which is known as its constituent, had significantly anti-lipid peroxidative effects.s.