• Title/Summary/Keyword: Liver enzyme

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Production of Lignin Peroxidase by Phellinus igniarius and Cytotoxic Effects of Lignin Hydrolysates Derived from Wood Biomass on Cancer Cells

  • Lee, Jae-Sung;Lee, Jong-Suk;Yoon, Jae-Don;Beak, Sung-Mok;Bosire, Kefa-O.;Lee, Yong-Soo;Kim, Jung-Ae
    • Biomolecules & Therapeutics
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    • v.12 no.3
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    • pp.189-193
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    • 2004
  • Over the past several years, research efforts have been directed both at economically producing valuable substances from the wood biomass and at producing lignolytic enzymes at a lower cost. In the present study, we found that Phellinus igniarius, the basidiomycetes, secreted lignin peroxidase as a main lignolytic enzyme, which was detected maximum activity at 16th day of culture and showed 37 kDa of molecular mass in identification by activity assay and purification by anion-exchange chromatography. The Phellinus igniarius-derived lignin peroxidase hydrolyzed steam-exploded wood (Quercus mongolica) powder into small molecules showing cytotoxicity against cancer cel1s (HepG2 hepatoma, SK-N-SH neuroblastoma, B16 melanoma, MBT-2 bladder cancer). In addition, the enzyme hydrlysates of lignins (ELg) that were extracted from the steam-exploded oak showed more potent cytotoxic effects on the cancer cells than the enzyme hydrolysates of wood biomass (EWp), indicating that the cytotoxic effect of EWp may be due to the enzyme-degraded products of lignin among the lignocellulosics. Furthermore, the cytotoxic effect of ELg on Chang, normal liver cells, was much less potent than that of ELg on HepG2 and B16 cancer cells, indicating that the cytotoxic effect of ELg may be specific for cancer cells. The present results suggest that Phellinus igniarius may be a useful resource for the large-scale production of lignin peroxidase and that the lignin peroxidase may be applied for the generation of valuable biodegradation products from wood lignocellulosics for medical use.

Effects of Silkworm(Bombyx mori L..) Powder on Lipofuscin, Acetylcholine and Its Related Enzyme Activities in Brain of Rats (뇌조직의 리포푸신, 아세틸콜린 및 그 관련효소 활성에 미치는 누에분말의 영향)

  • 최진호;김대익;박수현;김정민;조원기;이희삼;류강선
    • Journal of Life Science
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    • v.10 no.6
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    • pp.564-569
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    • 2000
  • This study was designed to investigate the effect of silkworm (Bombyx mori L.) powder on lipofuscin, acetylcholine (ACh) and its related enzyme activities in brain of rats. Sprague-Dawley (SD) male rats ($160{\pm}10g$) were fed basic diet (control group), and experimental diets (SWP-200 and SWP-400 groups) added 200 and 400 mg/kg BW/day for 6 weeks. In case of liver membranes, lipofuscin (LF) levels resulted in a slight decreases (4.6% and 11.5%, respectively) in SWP-200 and SWP-400 groups compared with control group. But in case of barin as the most sensitive organ, LF levels were remarkably inhibited about 16.7% and 20.0% in SWP-200 and SWP-400 groups compared with control group. There were no significant differences in acetylcholine (ACh) syntheses as a very important neurotransmitter, and choline acetyltransferase (ChAT) activities as a synthesis enzyme of ACh, and acetylcholinesterase (AChE) activities as a hydrolysis enzyme, which were concerned in transmission of neuron through synapses in brain of SWP-200 and SWP-400 groups compared with control group. Monoamine oxidase-B (MAO-B) activities were significantly inhibited (about 10.2%) in brain of SWP-400 groups compared with control group. These results suggest that inhibiting effects of LF accumulation and MAO-B activity of silkworm powder (SWP) may play a pivotal role in attenuating a various age-related changes for improvement of brain function.

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Effects of Silk Fibroin Powder on Lipofuscin, Acetylcholine and Its Related Enzyme Activities in Brain of SD Rats (뇌조직의 리포푸신, 아세틸콜린 및 그 관련효소 활성에 미치는 실크 피브로인의 영향)

  • 최진호;김대익;박수현;김동우;이광길;여주홍;김정민;이용우
    • Journal of Sericultural and Entomological Science
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    • v.42 no.2
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    • pp.120-125
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    • 2000
  • This study was designed to investigate the effects of silk fibroin (Mw 500) powder (SFP) on lipofuscin, acetylcholine (ACh) and its related enzyme activities in brain of rats. Sprague-Dawley (SD) male rats (160$\pm$10 g) were fed basic diet (control group), and experimental diets (SFP-2.5 and SFp-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. In case of liver membranes, lipofuscin (LF) levels resulted in a considerable decreases (11.5% and 13.8%, respectively) in SFP-2.5 and SFP-5.0 groups compared with control group. But in case of brain as the most sensitive organ, LF levels were remarkably inhibited about 18.3% and 21.7% in SFP-2.5 and SFP-5.0 groups compared with control group. Acetylcholine (ACh) levels were considerable decrease (3.0% and 9.2%, respectively) in brain membranes of SFP-2.5 and SFP-5.0 groups compared with control group. choine acetyltranferase (ChAT) activities as a synthesis enzyme of ACh, and acetylcholinesterase (AChE) activities as a hydrolysis enzyme resulted in a slight increases (2.4% and 3.0%, 4.6% and 6.3%, respectively), but significance difference between ChAT and AChE activities by SFP administration could be not obtained. Monoamine oxidase-B (MAO-B) activities were significantly inhibited (9.5% and 12.6%, respectively) in brain of SEP-2.5 and SFP-5.0 groups compared with control group. These results suggest that inhibiting effects of LF accumulation and MAO-B activity of silk fibroin(SFP) may play a pivotal role in protecting learning memory impairments by attenuating a various age-related changes for improvement of brain function.

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Development of Fecal Microbial Enzyme Mix for Mutagenicity Assay of Natural Products

  • Yeo, Hee-Kyung;Hyun, Yang-Jin;Jang, Se-Eun;Han, Myung-Joo;Lee, Yong-Sup;Kim, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.838-848
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    • 2012
  • Orally administered herbal glycosides are metabolized to their hydrophobic compounds by intestinal microflora in the intestine of animals and human, not liver enzymes, and absorbed from the intestine to the blood. Of these metabolites, some, such as quercetin and kaempherol, are mutagenic. The fecal bacterial enzyme fraction (fecalase) of human or animals has been used for measuring the mutagenicity of dietary glycosides. However, the fecalase activity between individuals is significantly different and its preparation is laborious and odious. Therefore, we developed a fecal microbial enzyme mix (FM) usable in the Ames test to remediate the fluctuated reaction system activating natural glycosides to mutagens. We selected, cultured, and mixed 4 bacteria highly producing glycosidase activities based on a cell-free extract of feces (fecalase) from 100 healthy Korean volunteers. When the mutagenicities of rutin and methanol extract of the flos of Sophora japonica L. (SFME), of which the major constituent is rutin, towards Salmonella typhimurium strains TA 98, 100, 102, 1,535, and 1,537 were tested using FM and/or S9 mix, these agents were potently mutagenic. These mutagenicities using FM were not significantly different compared with those using Korean fecalase. SFME and rutin were potently mutagenic in the test when these were treated with fecalase or FM in the presence of S9 mix, followed by those treated with S9 mix alone and those with fecalase or FM. Freeze-dried FM was more stable in storage than fecalase. Based on these findings, FM could be usable instead of human fecalase in the Ames test.

Polymerization of ADP-Ribose Pyrophosphatase: Conversion Mechanism of $Mg^{2+}-Dependent$ ADP-Ribose Pyrophosphatase into $Mg^{2+}-Independent$ Form

  • Kim, Dae-Ki;Kim, Jong-Hyun;Song, Eun-Kyung;Han, Myung-Kwan;Kim, Jong-Suk
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.826-831
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    • 2003
  • ADP-ribose pyrophosphatase (ADPRase) hydrolyzes ADP-ribose (ADPR) into AMP and ribose-5'-phosphate. It is classified into two groups, $Mg^{2+}$-dependent and $Mg^{2+}$-independent ADPRase, depending on its $Mg^{2+}$requirement. Here, we purified $Mg^{2+}$-dependent ADPRase from rabbit liver and examined what factors affect $Mg^{2+}$ requirement. The purified enzyme showed a single band with the molecular weight of 34 kDa on SDS-PAGE both in the presence and absence of 2-mercaptoethanol. The molecular weight of the native enzyme calculated by gel filtration was 68 kDa, indicating that ADPRase is a dimer made up of two identical subunits. $Mg^{2+}$-dependent ADPRase with the highest ADPR affinity had a $K_m$ of 160$\pm$10 $\mu$M and a pH optimum of around pH 9.5. Treatment of the purified ADPRase with heated cytosol fractions at 37$^{\circ}C$ for 3 h caused some changes in the chemical properties of the enzyme, including an increase in molecular weight, a decrease in solubility, and a loss of $Mg^{2+}$-dependency. The molecular weight of the cytosol-treated ADPRase measured by gel filtration was over 420 kDa, suggesting, for the first time, that ADPRase could be polymerized by undefined cytoplasmic factors, and that polymerization is accompanied by changes in the solubility and metal ion dependency of the enzyme.

Ursodeoxycholic Acid in the Prevention of Pediatric Parenteral Nutrition-associated Cholestasis (소아 총정맥영양의 간담도계 합병증에 대한 Ursodeoxycholic acid의 예방효과에 대한 연구)

  • Kim, Ji Hee;Min, Myung Sook;In, Yong Won;Shon, Kie Ho;Choi, Kyung Eob;Choe, Yon Ho;Beck, Nam Sun;Lee, Suk Hyang;Park, Tae Sung
    • Korean Journal of Clinical Pharmacy
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    • v.15 no.1
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    • pp.9-20
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    • 2005
  • Cholestatic liver disease is a frequent complication of prolonged parenteral nutrition, especially in premature infants. Numerous factors have been cited as contributing to TPN associated cholestasis. However the exact etiology remains obscure. Ursodeoxycholic acid (UDCA) has been reported to be beneficial far children and adults with various chronic cholestatic liver disease. The aim of this prospective, randomized, double-blind, placebo-controlled study was to determine the preventive effects of UDCA administration during TPN. Seventeen pediatric patients (8 boys and 9 girls) undergoing TPN were assigned randomly to two groups, UDCA and placebo group. UDCA group (n=9) received 15 mg/kg/day UDCA and placebo group (n=8) received 15 mg/kg/day placebo enterally during the TPN period. Liver function tests (total bilirubin, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase) were per-formed before TPN and weekly or three times a week. The patients' weights, complete blood count, composition of TPN, and the infusion rate of TPN and lipid were monitored everyday. Calcium and phosphate were monitored twice a week. Between the UDCA and placebo groups, there were no differences in weight at the onset of TPN, birth weight, duration of TPN, respiratory distress syndrome associated with prematurity, age at the onset of TPN, gestational age, the number of days the patients received antibiotics, the number of patients received enteral nutritions and the composition of TPN. In contrast, there was a significant difference between the UDCA and placebo groups in alanine aminotransferase levels during TPN. It doesn't seem that UDCA administration during TPN correlates directly with improvement of liver function. But the preventive administration of UDCA may be effective in reducing liver enzyme, alanine aminotransferase and has no adverse effects.

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Protective Effect of the Herbal Combination HCE-2 on Alcohol Metabolism and Alcohol-induced Liver Injury in Ethanol-loaded Rats (한약 복합추출물 HCE-2가 알코올을 투여한 쥐의 알코올 대사 및 간기능에 미치는 영향)

  • Han, Dong-Oh;Park, Soo-Jin;Sur, Bong-Jun;Lee, Hye-Jung;Kim, Sung-Hoon;Hahm, Dae-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.5
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    • pp.1081-1086
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    • 2007
  • This study was performed to investigate the medicinal effects of the herbal combination extracts-2 (HCE-2), consisting of Artemisia capillaris Thunb., Lonicera japonica Thunb., Prunella vulgaris var. lilacina, and Hovenia dulcis Thunb. on the alcohol-induced liver injury in rats. The rats were randomly divided into four groups: normal group (n =6), non-treated control group (n =6), saline-treated group (n =6) and the herbal combination extract (HCE-2)-treated group (n =6). The rats in the alcohol-loaded groups were orally administered with ethanol at a daily dose of 4 g/kg-body weight for 5 weeks. Thirty minutes before the ethanol injection, saline or herbal combination extracts was administered by using a gastrogavage. Blood and liver tissue samples were taken out from the hearts and livers of the rats, respectively, on 15th and 38th days. The activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using an enzyme-linked immunosorbent assay (ELISA). We also investigated the protective effect of the herbal combination extracts by Hematoxylin-Eosin staining on histological sections of rat liver. In this study, the oral administration of the herbal combination extracts significantly reduced the serum levels of AST and ALT, which had been raised by alcohol-induced liver injury. Histological analysis and apparent observation of liver also showed the preventive effect of the herbal combination extracts in a chronic alcohol-induced rat model. Theses results revealed that the herbal combination extracts effectively prevented hepatic damage consequent to the chronic exposure to repetitive administration of ethanol and could be used as a primary resource of a health beverage or herbal medicine, alleviating the alcohol-induced hepatic injury and hangover symptoms.

Effects of Water Extract from Orostachys japonicus on Lipid and Antioxidant Metabolism in the Liver of Obese Mice (와송이 고지방 식이로 유도된 비만 쥐의 간내 지질 및 항산화 대사에 미치는 영향)

  • Lee, Hyeong-Seon;Kim, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.515-521
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    • 2019
  • The purpose of this study was to investigate the changes in the liver lipid and antioxidant metabolism in obese rats following ingestion of Orostachys japonicus (OJ) hot water extract. The rats were fed a high fat diet for 8 weeks to induce obesity. In the experimental group, 2.5% or 5% OJ hot water extract was added to the diets for 6 weeks. To determine the changes in the liver lipid and antioxidant metabolism in obese rats, the livers were collected and weighed, lipid content was measured, and antioxidant enzyme activity was determined. A significant decrease in liver weight was observed in the group fed with OJ hot water extracts and decreased lipid accumulation was observed in the morphological observation of liver tissue. Total cholesterol and triglyceride contents were also decreased. Both OJ extracts significantly decreased 2,2-diphenyl-1-picrylhydrazyl radical and intracellular reactive oxygen species levels. Further, the levels of the antioxidant enzymes superoxide dismutase and catalase were significantly increased in rats treated with OJ extracts. These results suggest that the intake of OJ hot water extracts may improve liver lipid metabolism by activating antioxidant enzymes.

Functionalities of Squid Liver Hydrolysates (오징어 간 가수분해물의 기능성)

  • Lee, Su-Seon;Park, Si-Hyang;Park, Joo-Dong;Konno, Kunihiko;Choi, Yeung Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.12
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    • pp.1677-1685
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    • 2012
  • The autolysate and hydrolysate of a common squid liver, Todarodes pacificus, were prepared. Autolysis (liver ratio, pH, temperature) and Protamex-treated hydrolysis (pH, temperature, ratio of protease to liver) conditions were optimized by response surface methodology using central composite design for under 1 hr of hydrolysis time. The desirability profile indicated that maximum DH could be achieved at a squid liver of 93.5%, pH 6.4, and $47^{\circ}C$ in autolysis, while that of Protamex-treated hydrolysis did at a Protamex-to-squid liver level of 0.33%, pH 6.0, and $55^{\circ}C$. Three amino acids, proline, cysteine, and methionine, were not detected in the total amino acid composition of the Protamex-treated hydrolysate, while they were detected in the free amino acid composition. Cadmium was $8.32{\pm}0.03$ mg/100 g-powder for raw, $3.56{\pm}0.02$ mg/100 g-powder for the autolysate, and $13.26{\pm}0.04$ mg/100 g powder for the Protamex-treated hydrolysate. The major molecular weight ranged from 1.0 to 1.5 kDa for the autolysate and from 210 to 470 Da for the Protamex-treated hydrolysate. Food functionalities of the autolysate, such as surface hydrolphobicity, emulsion activity index, emulsion stability, water, and fat adsorption, were similar to the Protamex-treated hydrolysate. Both the autolysate and Protamex-hydrolysate showed high inhibitory activities on the angiotensin-I converting enzyme. Cell toxicity against the HepG2 cell line was not detected in the autolysate or the Protamex-treated hydrolysate by 200 ${\mu}g/mL$.

Effect of Cold Exposure on Thyroid Thermogenesis in Rats (한냉에 노출된 흰쥐에서 갑상선 호르몬이 체열 생산인 미치는 영향)

  • 황애란
    • Journal of Korean Academy of Nursing
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    • v.13 no.2
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    • pp.87-104
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    • 1983
  • It has been well documented that animals exposed to cold show increased activity of thyroid gland. The calorigenic action of thyroid hormone has been demonstrated by a variety of in vivo and in vitro studies. According to Edelman et al., the thyroid thermogenesis is due to activation of energy consuming processes, especially the active sodium transport by the hormone in target tissues. If so, the increase in thyroid activity during cold exposure should induce increased capacity of sodium transport in target tissue and the change in tissue metabolism should be precisely correlated with the change in Na+_K+_ATPase activity of the tissue. This possibility was tested in the present study: in one series, changes in oxygen consumption and Na+_K+_-ATPase activity of liver preparations were measured in rats as a function of thyroid status, in order to establish the effect of thyroid hormone on the tissue respiration and enzyme system in another series, the effect of cold stimulus on the serum thyroid hormone level, hepatic tissue oxygen consumption and Na+_K+_ATPase activity in rats. The results obtained are as follows: 1. The Na+_dependent oxygen consumption of liver slices, the oxygen consumption of liver mitochondria and the Na+_K+_ATPase activity of liver preparations were significantly inhibited in hypothyroidism and activated in hyperthyroidism. Kinetic analysis indicated that the Vmax. of Na+_K+_ATPase was decreased in hypothyroidism and increased in hyperth)'roidism. 2. In cold exposed rats, the serum triiodothyronine (T₃) level increased rapidly during the initial one day of cold exposure, then declined slowly to the control level after two weeks. The serum thyroxine (T₄) level decreased gradually throughout the cold exposure. Accordingly the T₃/T₄ratio increased. The mitochondrial oxygen consumption and the Na+_dependent oxygen consumption of liver slices increased during the first two days and then remained unchanged thereafter The activity of the Na+_K+_ATPase in liver preparations increased during cold exposure with a time course similar to that of oxygen consumption. Kinetic analysis indicated that the Vmax. of Na+_K+_ATPase increased. 3. Once the animal was adapted to cold, induction of hypothyroidism did not significantly alter the hepatic oxygen consumption and Na+_K+_ATPase activity. These results indicate that: 1) thyroid hormone increases capacities of mitochondrial respiration and active sodium transport in target tissues such as liver; 2) the increased T₃level during the initial period of cold exposure facilitates biosynthesis of Na+_K+_ATPase and mitochondrial enzymes for oxidative phosphorylation, leading to enhanced production and utilization of ATP, hence heat production.

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