• Title/Summary/Keyword: Glutathione S transferase

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Effects of Sardine Oil Fedding and Vitamin E Supplementation on Histopathological Changes and $\alpha$-L-Fucosidase Activity in Experimental Hepatocarcinogenesis (정어리유 섭취와 비타민 E 보충이 실험적 간 발암과정의 병리조직학적 변화와 $\alpha$-L-fucosidase의 활성도에 미치는 영향)

  • 김정희;윤혜진
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.119-126
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    • 2001
  • Effects of Sardine Oil Feeding and Vitamin E Supplementation on Histopathological Changes and $\alpha$-L-fucosidase activity in experimental hepatocarcinogenesis. Sprague-Dawley rats weighing 80~90 g were fed the diet containing either 15% corn oil (CO) or sardine oil (SO) with or without vitamin E supplements (dl-$\alpha$-tocopherol acetate 800 IU/kg diet) for 8 weeks. After 2 weeks of feeding, the rats were given a single intraperitoneal injectin of diethylnitrosamine (DEN, 200 mg/kg BW). From the fifth week, rats were given 0.02% acetylaminofluorene (AAF) in diet for 4 weeks. At the seventh week, 0.05% phenobarbital in liver and hepatic glutathione S-transferase palcental form positive (GST-P+) foci were examined by Hematoxylin& Eosin (H&E) staining and immunohistochemical method, respectively. Serum $\alpha$-L-fucosidase activity was determined. The livers fromt he carcinogen treated rats showed significantly increased formation of GST-P+ foci at sacrifice points while the livers fromthe non-carcinogen treated groups showed almost no foci. Although GST-P+ foci formation was not affected by dietary oil, it was increased unexpectedly by vitamin E supplementation. Histopathological changes were similar to patterns of GST-P+ foci formation in almost all groups. Serum $\alpha$-L-fucosidase activities were increased by carcinogen treatment in all dietary groups. $\alpha$-L-fucosidase activities were positively correlated with GST-P+ foci formation. There results suggest that excessive vitamin E supplementation can enhance hepatocarcinogenesis although the mechanisms involved are not clearly understood.

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Heat Shock Protein $90{\beta}$ Inhibits Phospholipase $C{\gamma}-1$ Activity in vitro

  • Cho, Sang-Min;Kim, Sung-Kuk;Chang, Jong-Soo
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.419-425
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    • 2006
  • Phospholipase $C-{\gamma}1\;(PLC-{\gamma}1)$ is an important signaling molecule for cell proliferation and differentiation. $PLC-{\gamma}1$ contains two pleckstrin homology (PH) domains, which are responsible for protein-protein interaction and protein-lipid interaction. $PLC-{\gamma}1$ also has two Src homology (SH)2 domains and a SH3 domain, which are responsible for protein- protein interaction. To identity proteins that specifically binds to PH domain of $PLC-{\gamma}1$, we prepared and incubated the glutathione S-transferase(GST)-fused PH domains of $PLC-{\gamma}1$ with COS7 cell lysate. We found that 90 kDa protein specifically binds to PH domain of $PLC-{\gamma}1$. By matrix-assisted laser desorption ionization time of flight-mass spectrometry, the 90 kDa protein revealed to be heat shock protein (Hsp) $90{\beta}$. Hsp $90{\beta}$ is a molecular chaperone that stabilizes and facilitates the folding of proteins that are involved in cell signaling, including receptors for steroids hormones and a variety of protein kinases. To know whether Hsp $90{\beta}$ affects on $PLC-{\gamma}1$ activity, we performed $PIP_2$ hydrolyzing activity of $PLC-{\gamma}1$ in the presence of purified Hsp $90{\beta}$ in vitro. Our results show that the Hsp $90{\beta}$ dose-dependently inhibits the enzymatic activity of $PLC-{\gamma}1$ and further suggest that Hsp $90{\beta}$ regulates cell growth and differentiation via regulation of $PLC-{\gamma}1$ activity.

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A Simple ELISA for Screening Ligands of Peroxisome Proliferator-activated Receptor γ

  • Cho, Min-Chul;Lee, Hae-Sook;Kim, Jae-Hwa;Choe, Yong-Kyung;Hong, Jin-Tae;Paik, Sang-Gi;Yoon, Do-Young
    • BMB Reports
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    • v.36 no.2
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    • pp.207-213
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    • 2003
  • Peroxisome proliferator-activated receptors (PPARs) are orphan nuclear hormone receptors that are known to control the expression of genes that are involved in lipid homeostasis and energy balance. PPARs activate gene transcription in response to a variety of compounds, including hypolipidemic drugs. Most of these compounds have high affinity to the ligand-binding domain (LBD) of PPARs and cause a conformational change within PPARs. As a result, the receptor is converted to an activated mode that promotes the recruitment fo co-activators such as the steroid receptor co-activator-1 (SRC-1). Based on the activation mechanism of PPARs (the ligand binding to $PPAR{\gamma}$ induces interactions of the receptor with transcriptional co-activators), we performed Western blot and ELISA. These showed that the indomethacin, a $PPAR{\gamma}$ ligand, increased the binding between $PPAR{\gamma}$ and SRC-1 in a ligand dose-dependent manner. These results suggested that the in vitro conformational change of $PPAR{\gamma}$ by ligands was also induced, and increased the levels of the ligand-dependent interaction with SRC-1. Collectively, we developed a novel and useful ELISA system for the mass screening of $PPAR{\gamma}$ ligands. This screening system (based on the interaction between $PPAR{\gamma}$ and SRC-1) may be a promising system in the development of drugs for metabolic disorders.

The Effects of Houttuynia cordata on the Hepatic Bromobenzene Metabolizing Enzyme System in Rats and Isolation of Phenolic Compounds (흰쥐의 브로모벤젠대사계에 미치는 어성초의 영향과 페놀성 화합물의 분리)

  • Hur, Jong-Moon;Park, Ju-Gwon;Park, Sung-Jong;Lee, Jong-Ho;Sung, Nak-Ju;Choi, Myeong-Rak;Song, Sang-Ho;Kim, Moon-Sung;Choi, Jong-Won;Park, Jong-Cheol
    • Korean Journal of Pharmacognosy
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    • v.31 no.2
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    • pp.228-234
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    • 2000
  • Effects of Houttuynia cordata on the level of lipid peroxide and the enzyme activities of the liver were investigated in bromobenzene-induced rats. Lipid peroxide content in liver was increased by bromobenzene. It was decreased when the methanol extract from the aerial parts of H. cordata was treated to the rat. The methanol extract reduced the activities of aminopyrine N-demethylase and aniline hydroxylase that increased by bromobenzene, however did not affect glutathione S-transferase activity. The methanol extract recovered the activity of epoxide hydrolase activity that decreased significantly by bromobenzene. We suggest that under our experimental conditions the extract might play an important play in the prevention of hepatotoxicity by reduction of aminopyrine N-demethylase and aniline hydroxylase activities as well as enhancement of epoxide hydrolase activity. Six phenolic compounds have been isolated from H. cordata and identified by means of spectral analysis as protocatechuic acid, quercetin, apigenin, afzelin, hyperoside and quercitrin.

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The Antitumor Effect in Sarcoma-180 Tumor Cell of Mice Administered with Japanese Apricot, Garlic or Ginger Doenjang (매실, 마늘 및 생강첨가 된장을 투여한 쥐의 Sarcoma-180 종양세포에서 항암효과)

  • Park, Kun-Young;Lee, Soo-Jin;Lee, Kyeoung-Im;Rhee, Sook-Hee
    • Korean journal of food and cookery science
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    • v.21 no.5
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    • pp.599-606
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    • 2005
  • The study was carried out to measure the antitumor effect of traditional doenjang (Korean soybean paste) added with Japanese apricot, garlic or ginger. Four kinds of traditional doenjang inhibited significantly the tumor growth in mice injected sarcoma-I80 cells. Especially, traditional doenjang added with ginger (Gi-TD) showed an inhibition of tumor cell activity of 97% by the administration of 1.0 mg/kg methanol extracts. Among Balb/c mouse administered doenjang extracts, the liver weight of mice fed Gi-TD was heavier than that of the group not administered doenjang. However, no difference was found between the control and doenjang administrated groups in weights of body, spleen, kidney and heart. The activity of natural killer (NK) cells was relatively high in mice administrated with the four kinds of doenjang. Particularly, mice administrated with the Gi-TD methanol extracts showed a strong activity of 82.9%. The activity of glutathione S-transferase (GST) in mice administrated with the 4 kinds of doenjang was higher than that of the group not administered with doenjang. In particular, the GST activity was the strongest in the group administrated with Gi-TD. The results suggest that Gi-TD has a strong growth inhibition activity against sarcoma-180 tumor cells.

GSTP1 Gene Ile105Val Polymorphism Causes an Elevated Risk for Bladder Carcinogenesis in Smokers

  • Pandith, Arshad Ahmad;Lateef, Adil;Shahnawaz, Sheikh;Hussain, Aashaq;Malla, Tahir Mohiuddin;Azad, Niyaz;Shehjar, Fahim;Salim, Mosin;Shah, Zafar Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6375-6378
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    • 2013
  • Background: The glutathione S transferase (GST) family of enzymes plays a vital role in the phase II biotransformation of environmental carcinogens, pollutants, drugs and other xenobiotics. GSTs are polymorphic and polymorphisms in GST genes have been associated with cancer susceptibility and prognosis. GSTP1 is associated with risk of various cancers including bladder cancer. A case control study was conducted to determine the genotype distribution of GSTP1 A>G SNP, to elucidate the possible role of this SNP as a risk factor in urinary bladder cancer (UBC) development and to examine its correlation with clinico-pathologic variables inUBC cases. Materials and Methods: Using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) approach, we tested the genotype distribution of 180 bladder cancer patients in comparison with 210 cancer-free controls from the same geographical region with matched frequency in age and gender. Results: We did not observe significant genotype differences between the control and bladder cancer patients overall with an odds ratio (OR)=1.23 (p>0.05). The rare allele (AG+GG) was found to be present more in cases (28.3%) than in controls (24%), though the association was not significant (p<0.05). However, a significant risk of more than 2-fold was found for the variant allele (AG+GG) with smokers in cases as compared to controls (p>0.05). Conclusions: Thus, it is evident from our study that GSTP1 SNP is not implicated overall in bladder cancer, but that the rare, valine-related allele is connected with higher susceptibility to bladder cancer in smokers and also males.

Molecular characterization and expression of cytosolic OASTL control cysteine metabolism in Mimosa pudica L.

  • Harun-Ur-Rashid, Md.;Masakazu, Fukuta;Hossain, Md. Amzad;Oku, Hirosuke;Iwasaki, Hironori;Oogai, Shigeki;Anai, Toyoaki
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.224-224
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    • 2017
  • In plants, cysteine(Cys) is decisive for protein and glutathione that acts as an indispensable sulfur grantor for methionine and many other sulfur containing secondary products. Cys formation is involved in the consecutive two reactions using two enzymes-serine acetyl transferase (SAT) and O-acetylserine (thiol)lyase (OASTL) and appeared in plant cytosol, chloroplast and mitochondria. OASTL is able to produce mimosine with 3-hydroxy-4-pyridone (3H4P) in lieu of $H_2S$ for Cys. In this report, we describe the first time cloning, purification and characterization of cytosolic(cy)OASTL from M. pudica and its expression in Escherichia coli and try to find out the cross link between this OASTL and the mimosine formation and to elucidate the metabolic role of cy-OASTL in M. pudica. The purified recombinant protein was 34.7 KDa. The optimum reaction pH and temperature was 6.5 and $50^{\circ}C$, respectively. The Michaelis constant (Km) and the Vmax value of the enzyme was $252{\pm}25{\mu}M$ and $57{\pm}3{\mu}M\;cysteine\;min^{-1}\;{\mu}g\;protein^{-1}$ for sulfide and $159{\pm}21{\mu}M$ and $58{\pm}2.4{\mu}M\;cysteine\;min^{-1}\;{\mu}g\;protein^{-1}$ for OAS subsequently. After cleaving the His-tag, we tried to observe cy-OASTL to form mimosine with appropriate substrate but it was not successful. It may be concluded that cy-OASTL of the present study is only Cys specific, not mimosine.

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Hepatoprotective Effect of the Methanol Fraction of Chinese Cabbage on Liver Injury in Rats Treated by bromobenzene (Chinese Cabbage 잎과 뿌리가 메탄올층의 Bromobenzene 간손상에 대한 보호효과)

  • Lee Hyo Jung;Kim Kwan Hyun;Lee Eun Ok;Choi Jong Won;Kang Kyung Sun;Yoon Byong Su;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.5
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    • pp.1177-1181
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    • 2003
  • Chinese cabbage is a vegetable of Cruciferous family. It was usually consumed as Kimchi. It was known to have amount of vitamin c. Recently the trend for the development of functional food combined with oriental herbs. For this aim the study was performed to evaluate the hepatoprotective effect via antioxidant activity of leaf and root of Sanchon Chinese cabbage(Brassica campetris L.) comparatively. The methanol extracts of Chinese cabbage were tested for investigating the effects on the formation of lipid peroxide and the activities of free radical generating enzyme in vitro in bromobenzene-treated rats. The methanol extracts of chinese cabbage reduced bromobenzene-induced hepatic lipid peroxidation and inhibited the activity of xanthine oxidase. The methanol extracts of chinese cabbage did not activate amionopyrine N-demethylase, aniline hydroxylase and glutathione S-transferase. Epoxide hydrolase activity was decreased by bromobenzene, which was restored by pretreatment of the methanol extracts of chinese cabbage. The results suggest that the methanol extracts of Chinese cabbage is reduced by enhancing the activity of epoxide hydrolase.

Expression and Antigenicity of Replicase Protein from Snow Mountain-Like Caliciviruses, Korean Isolates (한국형 사람 Calicivirus Replicase 단백의 발현 및 항원성 평가)

  • Chang, Mi-Yoon;Yang, Jai-Myung;Kim, Kyung-Hee
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.151-160
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    • 1997
  • In view of the potential of replicase protein as a diagnostic reagent for human caliciviruses (HuCVs), we have cloned and over-expressed this gene from the Snow Mountain-like Korean strains in Escherichia coli as a fusion protein with glutathione S-transferase (GST), and described the preliminary antigenic characterization of the recombinant products. Each 470bp fragment corresponding to highly conserved region of RNA-dependent RNA polymerase was generated by RT-PCR from stools of two diarrheal children, cloned in pMOSBlue T-vector, and subcloned between the EcoRI and SalI restriction sites of pGEX-4T-3, a GST gene fusion vector, yielding $pGCV_{pol}$. This construct expressed a Snow Mountain-like HuCV replicase under the control of the IPTG-inducible tac promoter. An extract prepared by sonication of the E. coli cell inclusion bodies bearing $pGCV_{pol}$ products was purified and analyzed by SDS-PAGE. After Coomassie blue staining, it was shown that the recombinant replicase migrated on the gels with an approximate molecular mass of 46.5 kDa, that was subsequently cleaved into a 26 kDa GST fragment and a 20.5 kDa replicase protein upon digestion with thrombin protease. The replicase was recognized on immunoblotting with the sera from symptomatic children with the HuCV-associated diarrhea but not by asymptomatic sera from adults. The results presented the first biological activity of individually expressed HuCV replicase subunit and provided important reagents for diagnosis of HuCV infection.

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Modifying Effects of Ellagic Acid in Food on Carcinogenesis (식품 중 Ellagic acid의 발암수식효과)

  • 장동덕;신동환;홍충만;조재천;한정희
    • Journal of Food Hygiene and Safety
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    • v.13 no.1
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    • pp.29-33
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    • 1998
  • The effect of ellagic acid (EA) on hepatocarcinogenesis induced by diethylnitrosamine (DEN), and promoted by phenobarbital (PB), and hepatectomized partially was investigated in male Wi star rats. All rats were injected 200 mg of DEN intraperitoneally, received 0.05 % of PB in drinking water at week 2, and hepatectomized 2/3 of liver at week 3. Rats of group 2, 3 and 4 were fed diet containing 400ppm EA for 1 week before DEN administration, for 9 weeks from beginning of experiment to sacrifice and for 6 weeks from PB treatment to sacrifice respectively. Rats of group 5, 6 and 7 were fed 800 ppm EA in the same manner as group 2, 3 and 4. Animals were killed at 8 weeks after DEN administration. The number and area of preneoplastic lesions were quantified the glutathione-S-transferase placental-form (GST-P) positive foci using immunohistochemical method. Decrease of number and area of the positive foci was observed in the rats fed 400 ppm EA for 9 weeks. In addition, the reduction of the foci can examine in all group fed 800 ppm EA. In conclusion, EA inhibited the hepatocarcinogenesis induced by DEN when it was administrated 800 ppm.

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