• Title/Summary/Keyword: DEHYDROGENASE

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The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture (양파(Allium cepa) 추출물의 간보호 및 항산화 효과)

  • Rhim, Tae-Jin;Lim, Sang-Cheol
    • Proceedings of the Plant Resources Society of Korea Conference
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    • v.18 no.1
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    • pp.52-60
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    • 2005
  • The objectives of present study were to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tert-butyl hydroperoxide(t-BHP), potent oxidizing agent for liver injury for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring catalase, glutathione peroxidase(GSH-Px), glutathione reductase(GSH-Rd) activities as well as DNA strand breaking assay. Incubation with t-BHP alone increased GOT and LDH activities and TBARS concentration but decreased MTT reduction. Onion extracts at the concentration of 0.05 mg/ml began to decrease GOT and LDH activities induced by 1.5 mM t-BHP. Decreased MTT reduction began to be increased by onion extract at the concentration of 0.01 mg/ml. Onion extracts at the concentration of 0.01 mg/ml began to decrease TBARS concentration induced by t-BHP. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, GSH-Px and GSH-Rd activities of hepatocytes were significantly decreased by 1.5 mM t-BHP for 1 hr incubation. Onion extracts, on the other hand, at the concentration of 0.1 mg/ml began to prevent t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton regents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition in lipid peroxidation.

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The Effect of Treponema Denticola and Treponema Lecithinolyticum on Periodontal Ligament Cells (Treponema Denticola와 Treponema Lecithinolyticum이 치주인대세포에 미치는 영향)

  • Jung, Jung-Hag;Choi, Bong-Kyu;Moon, Ik-Sang;Cho, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.29 no.2
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    • pp.311-326
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    • 1999
  • This study was investigated to observe the effect of Treponema denticola(TDC) and Treponema lecithinolyticum(TLC) on cultured human periodontal ligament cells. Several experiments were performed including MTT test for the inhibition effect of cell proliferation, LDH test for the cytotoxicity , gelatin zymography for the gelatinase activation and observation of cell morphology change using the phase-contrast microscopy. The results were as follows. 1. The effect of concentration on cell proliferation with time showed an inhibitory effect at high concentration $(150{\mu}g/well)$ for TLC and at low concentration( $9.4{\mu}gwell$ ) for TDC. 2. The effect of time on cell proliferation with concentration showed an inhibitory effect at $150{\mu}g/well$ on 2-day incubation for TLC and at $9.4{\mu}g/well$ on 2-day incubation for TDC. 3. The effect of heat-treated TDC and TLC on the inhibition of cell proliferation showed the difference in the heat-treated group compared to the non-heat treated group for TDC, whereas no difference was found for TLC. 4. The morphological changes which were observed from the phase-contrast microscopy showed the difference in the test group compared to the control group. The loss of spindle-like appearance, cell-to-cell detachment and inhibition of cell proliferation were observed. 5. There was no difference of the cytotoxicity effect between the test group and the control group in the LDH test. 6. The active form of progelatinase A with molecular weight 72kDa was activated in both TDC and TLC on the gelatin zymography. Regarding to the above results, TDC and TLC have an effect on periodontal ligament cells by playing an inhibitory role in cell proliferation and appears to activate progelatinase A which degrades type IV collagen.

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Antioxidant Effects of Eriodictyol on Hydrogen Peroxide-Induced Oxidative Stress in HepG2 Cells (산화스트레스가 유도된 HepG2 세포에서 Eriodictyol의 항산화 효과)

  • Joo, Tae-Woo;Hong, Sung-Hyun;Park, Sun-Young;Kim, Gur-Yoo;Jhoo, Jin-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.510-517
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    • 2016
  • This study was conducted to investigate the antioxidant and hepatoprotective effects of eriodictyol compound against hydrogen peroxide-induced oxidative stress in HepG2 cells by measuring expression levels of antioxidant enzymes, liver function index enzyme activities, and inhibitory effects against reactive oxygen species (ROS) production. HepG2 cell viability was assessed using 3-(4,5-dimethyl thiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. In the concentration range of $10{\sim}50{\mu}g/mL$, eriodictyol displayed over 98% cell viability in HepG2 cells. The effects of increased gene expression on hydrogen peroxide-induced oxidative stress were analyzed by monitoring antioxidant enzyme (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPx) gene expression levels using real-time PCR. Eriodictyol compound significantly increased gene expression levels of SOD, CAT, and GPx in a dose-dependent manner ($10{\sim}50{\mu}g/mL$). Hepatoprotective effects against hydrogen peroxide-induced oxidative stress were analyzed by monitoring glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) activities in HepG2 cell culture medium using a biochemistry analyzer. Eriodictyol compound significantly reduced GOT, LDH, and GGT activities in a dose-dependent manner in HepG2 cells. ROS level in HepG2 cells was analyzed by 2',7'-dichlorofluorescein fluorescence diacetate assay, and eriodictyol compound effectively reduced the intracellular ROS level in HepG2 cells. The results reveal that eriodictyol compound can be useful for development of effective antioxidant and hepatoprotective agents.

Effects of Ramie Leaves on Improvement of Lipid Metabolism and Antiobesity Effect in Rats Fed a High Fat/High Cholesterol Diet (모시잎이 고지방-고콜레스테롤 식이 흰쥐의 지질대사 개선 및 항비만 효과에 미치는 영향)

  • Lee, Jae-Joon;Park, Mi-Ran;Kim, Ah-Ra;Lee, Myung-Yul
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.83-90
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    • 2011
  • This study was performed to investigate the effects of ramie (Boehmeria nivea) leaf powder on improvements in lipid metabolism in serum, liver and adipose tissue, and the anti-obesity effect in rats fed a high fat/high cholesterol diet for 4 weeks to induce a hyperlipidemic and obese model rat. Male Sprague-Dawley rats weighting 210 g were divided into four groups; a normal diet group (N), a high fat/high cholesterol diet group (HFC), a high fat/high cholesterol diet with 5% ramie leaf powder group (HFC-RL), and a high fat/high cholesterol diet with 10% ramie leaf powder group (HFC-RH). The body weight gain increased with a high fat/high cholesterol diet, but gradually decreased in the ramie leaf powder fed groups compared with the HFC group. The liver index in the HFC group was highest among the four groups, although the difference was not significant compared with the ramie leaf powder fed groups. The adipose tissues weight in the HFC group was heavier than that of the N group, whereas those of groups fed ramie leaf powder decreased gradually. Alkaline phosphatase activity was not different between the HFC groups, but serum alkaline aminotransferase and lactate dehydrogenase activities decreased significantly after ramie leaf powder feeding. Levels of serum triglyceride, total cholesterol, LDL-cholesterol, atherogenic index and cardiac risk factors tended to decrease in the ramie leaf powder fed groups compared with the HFC group, whereas serum HDL-cholesterol level decreased in the HFC group and markedly increased in the ramie leaf powder fed groups. Levels of triglyceride and total cholesterol in the liver were significantly lower in the ramie leaf powder fed groups than in the HFC group. Levels of triglyceride and total cholesterol in adipose tissues were also lower in the ramie leaves powder fed groups than in the HFC group. The activities of heparinreleasable lipoprotein lipase (LPL) and total-extractable LPL in adipose tissues increased in the HFC group compared to that of the N group, but those of the ramie leaf powder fed groups decreased significantly. These results suggest that ramie leaf powder may improve lipid metabolism in serum, liver and adipose tissue and potentially reduce lipid storage.

Hepatoprotective effect of Ainsliaea acerifolia water extract on LPS/D-GalN-induced acute liver injury in human HepG2 cells (단풍취 열수 추출물의 HepG2 인간간세포의 LPS/D-Gal에 의해 유발된 급성 간 손상에 대한 간보호 효과)

  • Lee, Sun-Yeop;Han, Joon-Hee;Choi, Da-Hye;Hong, Min;Kwon, Tae-Hyung;Lee, Yong-Jin;Yu, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.52 no.5
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    • pp.476-481
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    • 2020
  • This study investigated the antioxidant and hepatoprotective effects of Ainsliaea acerifolia water extract (AAWE) on HepG2 cells. Five types of caffeoylquinic acid (CQA) were detected in AAWE, namely, 4,5-di-O-caffeoylquinic acid (4,5-DCQA; 11.16 mg/g), 3,4-di-O-caffeoylquinic acid (3,4-DCQA; 5.23 mg/g), 5-O-caffeoylquinic acid (5-CQA; 4.88 mg/g), 3,5-di-O-caffeoylquinic acid (3,5-DCQA; 3.51 mg/g), and 4-O-caffeoylquinic acid (4-CQA; 3.31 mg/g). AAWE exerted ABTS+ antioxidant effects, evidenced by polyphenol content and 2,2'2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH radical scavenging) activities. AAWE (300 ㎍/mL) treatment significantly decreased the activities of gamma glutamyl transferase (GGT), aspartate transaminase (AST), and lactate dehydrogenase (LDH) as compared to control and exerted protective effects against the increase in liver function index induced by lipopolysaccharide (LPS)/galactosamine (D-GalN) in HepG2 cells. In addition, the secretion of tumor necrosis factor (TNF)-α by HepG2 cells induced by LPS/D-GalN significantly increased in all treatment groups compared to that in the control. However, AAWE (100-300 ㎍/mL) treatment significantly decreased the secretion of TNF-α compared to that in the control. These results suggest that AAWE treatment reduces hepatotoxicity by increasing antioxidant activities, reducing GGT, AST, and LDH activities, and inhibiting TNF-α secretion.

Effects of Water in Extracts of Pueraria Radix on Serum Enzymes Activities and Hormone in Aluminum-Administeredrats (갈근 열수 추출물이 알루미늄을 투여한 흰쥐의 혈청 효소 활성도와 호르몬에 미치는 영향)

  • Han, Sung-Hee;Shin, Mee-Kyung
    • Journal of the Korean Society of Food Culture
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    • v.20 no.1
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    • pp.113-122
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    • 2005
  • This study was designed to investigates the effects of Korean pueraris radix water extract in Al(Aluminum) administered rats. Forty-eight male Sprague-Dawley rats weighing $100{\pm}10g$ were used for this experiment and divided into following 6 groups; control group, 3% pueraria radix in water extract group, 1000 and 2000ppm Al group, 1000 and 2000ppm Al group with 3% pueraria radix in water extract group. The Al administered rats were given 1000 and 2000 ppm of $Al_2(SO_4)_3$ disoved in the distilled water. The Al content in the rats tissue of Al administered group was lower than in the rats tissue of Al group with 3% pueraria radix in water extract group. Plasma levels of renin and aldosterone activity was increased by Al administration group, compared with 3% pueraria radix in water extract group and Al administred group. Glutamate oxaloacetate transaminase(GOT) and Glutamate pyruvate transaminase(GPT) were increased in Al-administered group and lower in the 3% extracts of pueraria radix in water extract group. Lactate dehydrogenase(LDH) was lower in the 3% extracts of pueraria radix-Al group than in the Al group. This results suggested that pueraria radix in water extract group has a lowering effects on the accumulation of Al and it is belived that the pueraria radix in extracted water group has some protective effects to Al administered in rats, but the mechanism of these effects was obscure.

Development of an Analytical Method for Fluxapyroxad Determination in Agricultural Commodities by HPLC-UVD (HPLC-UVD를 이용한 농산물 중 Fluxapyroxad 잔류분석법 개발)

  • Kwon, Ji-Eun;Kim, HeeJung;Do, Jung-Ah;Park, Hyejin;Yoon, Ji-Young;Lee, Ji-Young;Chang, Moon-Ik;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.29 no.3
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    • pp.234-240
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    • 2014
  • Fluxapyroxad is classified as carboxamide fungicide that inhibits succinate dehydrogenase in complex II of mitochondrial respiratory chain, which results in inhibition of mycelial growth within the fungus target species. This study was carried out to assure the safety of fluxapyroxad residues in agricultural products by developing an official analytical method. A new, reliable analytical method was developed and validated using High Performance liquid Chromatograph-UV/visible detector (HPLC-UVD) for the determination of fluxapyroxad residues. The fluxapyroxad residues in samples were extracted with acetonitrile, partitioned with dichloromethane, and then purified with silica solid phase extraction (SPE) cartridge. Correlation coefficient($R^2$) of fluxapyroxad standard solution was 0.9999. The method was validated using apple, pear, peanut, pepper, hulled rice, potato, and soybean spiked with fluxapyroxad at 0.05 and 0.5 mg/kg. Average recoveries were 80.6~114.0% with relative standard deviation less than 10%, and limit of detection (LOD) and limit of quantification (LOQ) were 0.01 and 0.05 mg/kg, respectively. All validation parameters were followed with Codex guideline (CAC/GL 40). LC-MS (Liquid Chromatograph-Mass Spectrometer) was also applied to confirm the analytical method. Base on these results, this method was found to be appropriate fluxapyroxad residue determination and can be used as the official method of analysis.

Early Diagnosis of Fatty Liver-Hemorrhagic Syndrome using Blood Biochemistry in Commercial Layers (산란계의 혈액화학치 측정에 의한 지방간 출혈 증후군의 조기 진단)

  • So, H.H.;Jeon, E.O.;Byun, S.H.;Mo, I.P.
    • Korean Journal of Poultry Science
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    • v.36 no.2
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    • pp.165-175
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    • 2009
  • Fatty liver-hemorrhagic syndrome (FLHS) is a common nutritional disease in commercial layers and breeders. The most important clinical sign of FLHS is a sudden drop in egg production and increased mortality which causes significant economic loss in the poultry industry. However, the current diagnostic method for FLHS is based on the gross findings at necropsy which is not helpful to reduce the economic loss because of lateness of diagnosis. Therefore, we need early diagnosis and diagnostic methods before chickens were affected by FLHS. In this study we tried to evaluate the effectiveness of clinical pathology including blood chemistry as an early diagnostic method for FLHS in commercial chickens. Profiles of blood biochemistry were compared between two flocks selected in the same commercial layer farm based on the presence of FLHS clinical sings. A flock with clinical signs of FLHS was designated as FLHS and other flock without clinical signs of FLHS as Non-FLHS. Several parameters of blood biochemistry were selected and compared between FLHS and Non-FLHS to evaluate the possibility of early diagnosis. Average concentrations of serum cholesterol, serum calcium, aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and creatine kinase (CK) were $139.4\;{\pm}\;87.2$ (mg/dL), $24.5\;{\pm}\;5.4$ (mg/dL), $153.6\;{\pm}\;23.1$ (IU/L), $1238.3\;{\pm}\;475.2$ (IU/L) and $1107.3\;{\pm}\;422.8$ (IU/L) in Non-FLHS flock, respectively, and $210.2\;{\pm}\;173.2$ (mg/dL), $25.2\;{\pm}\;4.1$ (mg/dL), $174.3\;{\pm}\;53.5$ (IU/L), $1694.9\;{\pm}\;691.3$ (IU/L) and $1104.9\;{\pm}\;472.9$ (IU/L) in FLHS flock, respectively. The activities of serum cholesterol, AST and LDH except CK, were significantly higher in FLHS than those in Non-FLHS flock (p<0.05). Some birds of FLHS flock showed 2~17 times greater than in Non-FLHS flock. For the definitive diagnosis of FLHS in the flocks tested for blood chemistry, we analyzed fat content and histological lesion score in the liver sampled from both FLHS and Non-FLHS flock. Average liver fat contents based on dry weight were $16.1\;{\pm}\;0.4$ (%) in Non-FLHS flock and were $21.6\;{\pm}\;16.0$ (%) in FLHS flock. These result confirmed that FLHS flock was definitely affected by FLHS. The above results suggest that selected parameters of blood biochemistry, particularly AST, could be useful to diagnose FLHS before significant liver damage occurred in commercial layers.

Effects of Baicalin on Gene Expression Profiles during Adipogenesis of 3T3-L1 Cells (3T3-L1 세포의 지방세포형성과정에서 Baicalin에 의한 유전자 발현 프로파일 분석)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Chung, Sang-In;Cho, Soo-Hyun;Yoon, Yoo-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.1
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    • pp.54-63
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    • 2010
  • Baicalin, a flavonoid, was shown to have diverse effects such as anti-inflammatory, anti-cancer, anti-viral, anti-bacterial and others. Recently, we found that the baicalin inhibits adipogenesis through the modulations of anti-adipogenic and pro-adipogenic factors of the adipogenesis pathway. In the present study, we further characterized the molecular mechanism of the anti-adipogenic effect of baicalin using microarray technology. Microarray analyses were conducted to analyze the gene expression profiles during the differentiation time course (0 day, 2 day, 4 day and 7 day) in 3T3-L1 cells with or without baicalin treatment. We identified a total of 3972 genes of which expressions were changed more than 2 fold. These 3972 genes were further analyzed using hierarchical clustering analysis, resulting in 20 clusters. Four clusters among 20 showed clearly up-regulated expression patterns (cluster 8 and cluster 10) or clearly down-regulated expression patterns (cluster 12 and cluster 14) by baicalin treatment for over-all differentiation period. The cluster 8 and cluster 10 included many genes which enhance cell proliferation or inhibit adipogenesis. On the other hand, the cluster 12 and cluster 14 included many genes which are related with proliferation inhibition, cell cycle arrest, cell growth suppression or adipogenesis induction. In conclusion, these data provide detailed information on the molecular mechanism of baicalin-induced inhibition of adipogenesis.

Studies on the Radiation Induced Apoptosis by Morphological and Biochemical Analysis in A431 Cells (방사선에 조사된 인체세포주(A431)에서 형태학적 분석법과 생화학적 분석법을 이용한 방사선 유도 세포고사에 관한 연구)

  • Choi, Keun-Hee;Bom, Hee-Seung;Kim, Ji-Yeul
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.3
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    • pp.306-315
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    • 1999
  • Purpose: We performed this study to evaluate the process of radiation induced apoptosis in A431 skin epithelial cancer cell line. Materials and Methods: Low to high dose radiation (0, 2, 5, 10, 25 Gy) was given to A431 cells by Cs-137 cell irradiator. Apoptosis was evaluated by cell morphology, dye exclusion test, and DNA laddering. Results: Cell viability decreased as the radiation dose increased. Number of apoptotic bodies increased as radiation dose increased. It increased most significantly at 12 hours after irradiation. Lactate dehydrogenase activity in culture medium increased according to radiation dose and time after irradiation. DNA ladders could be identified in irradiated cells, but, it had no correlation with radiation dose or time after irradiation. Conclusion: Radiation-induced apoptosis which was the main course of cell death in A431 cells could be analyzed quantitatively by counting apoptotic bodies under microscope. Apoptosis increased as radiation dose increased.

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