• Title/Summary/Keyword: marker enzymes

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Hepatoprotective Activity of Salidroside Isolated from Acer Termentosum Max on D-galactosamine Induced Hepatotoxicity in Rats (산겨릅나무에서 분리한 salidroside의 간보호 효과)

  • Kim, Sung-Hoon;Park, Hee-Juhn;Choi, Jon-Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1525-1531
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    • 2008
  • Acer tegmentosum Max which is one of the specialized wildness medicinal herbs in gangwon province, has been widely used for hepatitis, liver cirrhosis, hepatic cancer, leukemia, diabetes mellitus, renal necrosis and edema, etc. In this study, the antioxidative and hepatoprotective effects of in vitro and in vivo were investigated in order to evaluate the possibility as hepatoprotective agents. Oral administration of methanol and butanol extact of Acer tegmentosum Max to d-galactosamine (D-GaIN) induced experimental liver injured rats was significantly reduced activities of marker enzymes(AST, ALT) and LDH activity in serum. Salidroside(Sal) isolated from the BuOH extract of Acer termentosum Max potenty showed the scavenzing effect on DPPH and inhibitory effect on lipid peroxidation. And significantly decrcease of MDA level in liver and activities of SOD GSH-Px and catalase were significantly improved by the treatment of Sal. Results of this study revealed that Sal could afford a significant protection in the alleviation of D-GaIN-induced hepatocellular injury.

Cloning and mutational analysis of pyrroquinoline quinone(PQQ) genes from a phosphate - solubilizing biocontrol bacterium Enterobacter intermedium.

  • Han, Song-Hee;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.94.2-95
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    • 2003
  • E. intermedium 60-2G possessing a strong ability to solubilize insoluble phosphate, has plant growth-promoting activity, induced systemic resistance activity against scab pathogen in cucumber, and antifungal activity against various phytopathogenic fungi. The phosphate solubilizing activity of 60-2G may be mainly accomplished by production of gluconic acid through a direct extracellular oxidation of glucose by glucose dehydrogenase that required a PQQ cofactor for its activation. A pqq gene cluster conferred Phosphate-solubilizing activity in E. coli DH5${\alpha}$ was cloned and sequenced. The 6,783 bP pqq sequence had six open reading frames (from A to F) and showed 50-95% homology to pqq genes from other bacteria. The E. coli strain expressing the pqq genes solubilized phosphate from hydroxyapatite after a pH drop to 4.0, which paralleled in time the secretion of gluconic acid. To study the role of PQQ in biocontrol traits of E. intermedium, PQQ mutants of 60-2G were constructed by marker exchangee mutagenesis. The PQQ mutants of E. intermedium were lost activities of solubilizing phosphate, growth inhibition of phytopathogenic fungi, and plant growth promotion. These findings suggest that PQQ plays an important role, possibly activation of certain enzymes, in several beneficial bacterial traits of E. intermedium by as yet an unknown mechanism.

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The Relative Identification of C. officinale and L. chuanxiong by PCR-Mediated Fingerprinting (천궁류(川芎類) 한약재의 유전자 감식 연구)

  • Choi, Ho-Young;Kim, Dong-Wook;Kim, Dong-Eun;Suh, Young-Bae;Ham, In-Hye
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.151-161
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    • 2005
  • Objectives : Our research purpose is to establish the standard identification analysis on C. officinale and L. chuanxiong in Korea and China by PCR-mediated fingerprinting. Methods : The Restriction Fragment Length Polymorphism (RFLP) and Randomly Amplified Polymorphic DNA (RAPD) method was used on Internal Transcribed Spacer (ITS) regions and rbcL regions to compare and discriminate genes extracted from crude drugs as C. officinale and L. chuanxiong in Korea and China. Results : L. chuanxiong Korea and China have very similar polymorphism, whereas L. chuanxiong in Korea and C. officinale have very different polymorphism in RFLP. And restriction enzymes AluI and SacI forms the specific fragment band only in C. officinale, they can be used as RFLP marker on ITS regions to discriminate among the species. Conclusions : The results could be applied in discriminating crude drugs among C. officinale and L. chuanxiong in Korea and China. Also they could be used in controlling drug quality, preserving medicinal plants, and improving plant description.

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Leukotriene Synthesis in Response to A23187 Is Inhibited by Methyl-β-Cyclodextrin in RBL-2H3 Cells

  • You, Hye Jin;Seo, Ji-Min;Moon, Ji-Young;Han, Sung-Sik;Ko, Young-Gyu;Kim, Jae-Hong
    • Molecules and Cells
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    • v.23 no.1
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    • pp.57-63
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    • 2007
  • Leukotrienes (LTs) are produced by several biosynthetic enzymes including cytosolic phospholipase $A_2$ ($cPLA_2$), 5-lipoxygenase (5-LO), and 5-lipoxygenase activating protein (FLAP) in the perinuclear area. In the present study, we showed that pretreatment with methyl-${\beta}$-cyclodextrin (MβCD), a cholesterol-depleting agent, dramatically reduced the synthesis of LTs in response to A23187 in mast cells. A23187-induced LT synthesis was inhibited by pretreatment with M${\beta}$CD, and this effect was reversed when cholesterol was added. In an approach to identifying the $M{\beta}CD$-sensitive protein(s), we observed that FLAP co-localized with flotillin-1, a lipid raft marker protein, in the lipid raft-rich low-density region of sucrose gradients. In addition, electron microscopic analysis revealed that FLAP co-localized with flotillin-1. Together, these results suggest that FLAP is present in cholesterol-rich lipid raft-like domains and that its localization in these domains is critical for LT synthesis.

Reversible Hepatic Toxic Effect of Crocin Dyes in Rats

  • Lin, Jen-Kun;Wang, Chau-Jong
    • Korean Journal of Pharmacognosy
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    • v.16 no.4
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    • pp.227-232
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    • 1985
  • Gardenia jasminodes has been medically used for anti-inflammation, sedation and anti-diarrhea; The extract of this plant has been traditionally used as food colorant and referred as crocin dyes. In the present study, the possible hepatic toxicity of this dye has been evaluated on the basis of its alteration on the marker enzymes, namely, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, alkaline phosphatase, lactate dehydrogenase and gamma-glutamyltransferase. Crocin dyes did not affect hepatic function when they were orally administered to rats in a daily dose of 50 mg/kg for 8 days, but could induce acute hepatic discoloration. A high dose of 100 mg/kg for 2 weeks could induce both hepatic damage and black pigmentation, but a lower dose of 10 mg/kg for 40 days did not The induced black pigmentation and the acute hepatic damage were completely reversible. In conclusion, the crocin dyes have a very low hepatic toxicity in rats, even in high experimental dosages which could hardly happen in human practice. It is therefore suggested that the crocin dyes are safe for coloring foods.

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Analysis of Gene Expression in 4,4'-Methylenedianiline-induced Acute Hepatotoxicity

  • Oh, Jung-Hwa;Yoon, Hea-Jin;Lim, Jung-Sun;Park, Han-Jin;Cho, Jae-Woo;Kwon, Myung-Sang;Yoon, Seok-Joo
    • Toxicological Research
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    • v.25 no.2
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    • pp.85-92
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    • 2009
  • 4,4'-Methylenedianiline (MDA) is an aromatic amine that is widely used in the industrial synthetic process. Genotoxic MDA forms DNA adducts in the liver and is known to induce liver damage in human and rats. To elucidate the molecular mechanisms associated with MDA-induced hepatotoxicity, we have identified genes differentially expressed by microarray approach. BALB/c male mice were treated once daily with MDA (20 mg/kg) up to 7 days via intraperitoneal injection (i.p.) and hepatic damages were revealed by histopathological observation and elevation of serum marker enzymes such as AST, ALT, ALP, cholesterol, DBIL, and TBIL. Microarray analysis showed that 952 genes were differentially expressed in the liver of MDA-treated mice and their biological functions and canonical pathways were further analyzed using Ingenuity Pathways Analysis (IPA). Toxicological functional analysis showed that genes related to hepatotoxicity such hyperplasia/hyperproliferation (Timp1), necrosis/cell death (Cd14, Mt1f, Timp1, and Pmaip1), hemorrhaging (Mt1f), cholestasis (Akr1c3, Hpx, and Slc10a2), and inflammation (Cd14 and Hpx) were differentially expressed in MDA-treated group. This gene expression profiling should be useful for elucidating the genetic events associated with aromatic amine-induced hepatotoxicity and for discovering the potential biomarkers for hepatotoxicity.

DNA Analysis of mtDNA COI Gene in the Sharp-toothed Eel (Muraenesox cinereus Forskal) from Yeosu, Jinhae, Jeju, Goseoung, Jangheung and Haenam Populations in Korea Using PCR-aided RFLP

  • Oh, Taeg-Yun;Jeong, Sun-Beom;Cho, Eun-Seob
    • Journal of Environmental Science International
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    • v.20 no.4
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    • pp.551-554
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    • 2011
  • The production of the sharp-toothed eel by commercial catch off waters of Korea is annually declined after 1978. This study was carried out to obtain the stock management of the sharp-toothed eel using the PCR-aided RFLP method. The mtDNA COI gene was amplified using species-specific primers and PCR product was observed to 700 bp. Amplified DNA fragments were treated with six kinds of restriction enzymes (BaeHI, EcoRI, PstI, Ksp22, HinfI and HaeIII). The treatment of HaeIII showed a distinct PCR product between Yeosu/Jinhae/Jeju/Goseoung and Jangheung/Haenam populations that were observed from 300 to 400 bp in reference to 100 bp molecular marker. However, DNA fragment within populations had an identical pattern. The phylogenetic homology is 82% between two populations inferred from RFLP PCR product pattern using NTsysPC ver. 2.1. The use of HaeIII plays an important role in discriminating populations. It is thought that adults after over-wintering in the southern part of Jeju migrate to the Yeosu, Jinhae and Goseoung regions to spawn instead of to southwestern waters. Individuals within populations showed a relatively active genetic mixing and migration regardless of geography. However, the genetic ancestor of Jangheung and Haenam populations is appeared to be more adjacent to China or Japan than Jeju.

Anticarcinogenic Effect and Modification of Cytochrome P450 2E1 by Dietary Garlic Powder in Diethylnitrosamine-Initiated Rat Hepatocarcinogenesis

  • Park, Kyung-Ae;Kweon, Sang-Hui;Choi, Hay-Mie
    • BMB Reports
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    • v.35 no.6
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    • pp.615-622
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    • 2002
  • The purpose of this study was to determine the effects of dietary garlic powder on diethylnitrosamine (DEN)-induced hepatocarcinogenesis and cytochrome P450 (CYP) enzymes in weaning male Sprague-Dawley rats by using the medium-term bioassay system of Ito et al. The rats were fed diets that contained 0, 0.5, 2.0 or 5.0% garlic powder for 8 weeks, beginning the diets with the intraperitoneal (i.p.) injection of DEN. The areas of placental glutathione S-transferase (GST-P) positive foci, an effective marker for DEN-initiated lesions, were significantly decreased in the rats that were fed garlic-powder diets; the numbers were significantly decreased only in the 2.0 and 5.0% garlic-powder diets. The p-nitrophenol hydroxylase (PNPH) activities and protein levels of CYP 2E1 in the hepatic microsomes of the rats that were fed the 2.0 and 5.0% garlic powder diet were much lower than those of the basal-diet groups. Pentoxyresorufin O-dealkylase (PROD) activity and CYP 2B1 protein level were not influenced by the garlic-powder diets and carcinogen treatment. Therefore, the suppression of CYP 2E1 by garlic in the diet might influence the formation of preneoplastic foci during hepatocarcinogenesis in rats that are initiated with DEN.

Effect of Swietenia macrophylla seeds on normal and alloxan-induced diabetic rats

  • A, Maiti;S, Dewanjee;SC, Mandal
    • Advances in Traditional Medicine
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    • v.7 no.4
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    • pp.363-371
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    • 2007
  • The methanol extract of the seed of Swietenia macrophylla (Meliaceae) seeds were investigated for its hypoglycaemic activity in normal and alloxan induced diabetic rats. Seed extract suspended in aqueous Tween 80 solution, the graded doses were administered to normal and experimental diabetic rats for 10 days. Fasting blood glucose levels, serum lipid profiles and changes in body weight were evaluated in normal rats while the hepatic marker enzymes and liver glycogen levels were evaluated additionally in diabetic rats. The diabetic groups treated with the seed extract were compared with standard drug glibenclamide. The findings showed the significant antidiabetic potential of the extract in ameliorating the diabetic conditions in diabetic rats. The extract also has hypoglycaemic action in normal rats. A toxicity study has been performed for the extract, which revealed that the extract is safe to use even at the doses of 3.2 mg/kg of body weight orally. The result from the present investigation indicates good correlation between the folk use and scientific investigation.

Phytochemical and pharmacological evaluation of the flowers of Sarcostemma brevistigma Wight

  • Lalitha, KG;Sethuraman, MG
    • Advances in Traditional Medicine
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    • v.9 no.3
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    • pp.252-258
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    • 2009
  • Shade-dried flowers of Sarcostemma brevistigma Wight Syn (SBF) belonging to Asclepiadaceae yielded a rare flavonol glycoside viz., Quercetin 5a prenyl, 3-O-glucosyl, 7-O-(4c-p-coumaroyl) neohesperidoside which were characterized based on chemical and spectral (including 2D NMR) studies. The ethyl acetate fraction of alcoholic concentrate (test sample) of flowers of this plant (SBF) was evaluated for its hepatoprotective and inhibition of lipid peroxidation activities to investigate the scientific basis of the traditional uses. The oral administration in varying doses viz., 125 and 250 mg/kg of aqueous suspension of SBF to rats for 7 days produced significant (P < 0.01) hepatoprotective effect comparable to that of standard drug silymarin. The SBF afforded good hepatoprotection against $CCl_4$ induced elevation levels of serum marker enzymes, serum bilirubin and liver weight. The free radical scavenging effects of SBF and flavonol glycoside (SA) were assigned by $Fe^{2+}$/ascorbate induction method (in vitro), which revealed the inhibition of lipid peroxidation. The SBF and SA showed prominent anti-lipid peroxidation activity ($IC_{50}$ about $\sim$180 mg/ml and 11.0 mg/ml respectively), which was comparable to standard drug curcumin ($IC_{50}$ about $\sim$8.25 mg/ml). Thus the hepatoprotective activity of SB could be correlated to the free radical scavenging property of the flavonol glycoside.