• Title/Summary/Keyword: H-transferase

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The overexpression of Arachis hypogaea resveratrol synthase 3 (AhRS3) modified the expression pattern of phenylpropanoid pathway genes in developing rice seeds

  • Lee, Choonseok;Jeong, Namhee;Kim, Dool-Yi;Ok, Hyun-Choong;Choi, Man-Soo;Park, Ki-Do;Kim, Jaehyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.105-105
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    • 2017
  • Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

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Effects of Long-Term Vitamin E and Butylated Hydroxytoluene Supplemented Diets on Murine Intestinal and Hepatic Antioxidant Enzyme Activities

  • Jang, I.S.;Chae, K.R.;Kang, T.S.;Kim, Y.K.;Kim, C.K.;Hwang, J.H.;Hwang, D.Y.;Choi, C.B.;Jung, K.K.;Cho, J.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.6
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    • pp.932-938
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    • 1999
  • The present study was designed to determine long-term feeding effects of vitamin E and BHT (butylated hydroxytoluene) on serum biochemical profiles, organ weight, and intestinal and hepatic antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and glutathione-S-transferase (GST) in ICR mice. Four wk old ICR mice (n=8 per group) were fed the diets supplemented with vitamin E (I ; 0.03% and II ; 0.3%) and BHT (I ; 0.05% and II ; 0.5%) for 12 months. Feeding the diets containing vitamin E and BHT had no effects on growth and serum biochemical profiles. However, feeding the diets supplemented with 0.5% BHT for 12 months significantly increased liver weight of the mice. In the small intestine, there were no effects of vitamin E or BHT on SOD and GSH-PX activities in the mucosa. However, the activity of intestinal GST of the mice that received 0.5% BHT was almost twice as high as that of control mice. In the liver, the activity of SOD was not affected by feeding antioxidants for 12 months, whereas GSH-PX activity was significantly increased in mice that received the diets containing BHT (0.05%, 0.5%) and vitamin E (0.03%, 0.3%). In addition, supplementation of 0.5% BHT markedly enhanced hepatic GST activity compared with other groups. Enhanced activity of GSH-PX in response to feeding vitamin E or BHT might aid hepatic enzymes to eliminate active oxygen in organs from mice. However, we could not exclude the possibility of increased lipid peroxidation by high dosage of BHT supplementation. More detailed study is necessary for assessment of preventive or toxicological effects of high dosage of BHT supplementation.

Cytoprotective Effect of Zinc-Mediated Antioxidant Gene Expression on Cortisol-Induced Cytotoxicity (Cortisol 유발 세포독성에 대한 아연 관련 항산화 유전자 발현 증가에 의한 세포보호 효과)

  • Chung, Mi Ja;Kim, Sung Hyun;Hwang, In Min
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.5
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    • pp.649-656
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    • 2015
  • The protective effect of zinc against cortisol-induced cell injury was examined in rainbow trout gill epithelial cells. Cells exposed to cortisol for 24 h showed increased leakage of lactate dehydrogenase (LDH) as well as decreased cell viability in a dose-dependent manner. Treatment with zinc ($100{\mu}M$ $ZnSO_4$) reduced the severity of both LDH release and cell death as well as protected cells against cortisol-induced caspase-3 activation, indicating reduction of apoptosis. Cortisol-induced cell death, leakage of LDH, and caspase-3 activation were blocked by the glucocorticoid receptor antagonist Mifepristone (RU-486), suggesting that cell injury was cortisol-dependent. In addition, we studied the effect of zinc on the expression of antioxidant genes such as metallothionein A (MTA), metallothionein B (MTB), glutathione-S-transferase (GST), and glucose-6-phosphate dehydrogenase (G6PD) during cortisol-induced cell injury. MTA, MTB, GST, and G6PD mRNA levels increased after treatment with zinc or cortisol, separately or in combination. Higher mRNA levels of MTA, MTB, GST, and G6PD were detected when cells were treated with $100{\mu}M$ $ZnSO_4$ and $1{\mu}M$ cortisol in combination at the same time compared to treatment with zinc or cortisol separately. Cells treated with zinc showed increased intracellular free zinc concentrations, and this response was significantly enhanced in cells treated with cortisol and zinc. In conclusion, zinc treatment inhibited cortisol-induced cytotoxicity and apoptosis through indirect antioxidant action.

Antihepatotoxic effect of Heat-treated Allium victorialis var. platyphyllum in $CCl_4-induced$ Rats and the Gas Chromatographic Analysis of Volatile Sulfur Substances

  • Park, Hee-Juhn;Jung, Hyun-Ju;Lim, Sang-Cheol;Jung, Won-Tae;Kim, Won-Bae;Park, Kwang-Kyun;Lee, Jin-Ha;Choi, Jong-Won
    • Natural Product Sciences
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    • v.11 no.2
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    • pp.67-74
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    • 2005
  • The ethanolic extracts of the leaves and bulbs of Allium victorialis var. platyphyllum (Liliaceae) collected from Daegwallyoung (D) and Ullung Island (U) in Korea were obtained using three different extracting methods. The first extracts, DL-1 DB-1, UL-1 and UB-1, were obtained from leaves (L) and bulbs (B) dried at $90^{\circ}C$, respectively, and the second extracts, DL-2, DB-2, UL-2 and UB-2, were obtained by extracting the leaves and bulbs of fresh plant parts. The third extracts DL-3, DB-3, UL-3 and UB-3 were obtained by incubating leaves and bulbs at $36^{\circ}C$. The six extracts obtained from A. victorialis var. platyphyllum at Daegwanllyoung (cultivated site) were orally administered to examine for a possible antihepatotoxic effect in $CCl_4-induced$ rats. DL-1 exhibited the most pronounced effect. The extracts inhibited serum ALT, AST, SDH, ${\gamma}-GT$, ALP and LDH activities elevated by $CCl_4$ injection and attenuated decreased glutathione S-transferase, glutatione reductase and ${\gamma}-glutamylcysteine$ synthetase activities and a decreased hepatic glutathione. However, the extracts obtained from Ullung Is. (native site) were less active than the extracts from Daegwallyoung, suggesting that A. victorialis var. platyphyllum from the cultivated site is more useful for functional food than of native site. These results also suggest that the antihepatotoxic effect is due to a higher content of hepatic glutathione. Gas chromatography of the twelve extracts showed significantly different sulfides, disulfides or trisulfides contents belonging to volatile sulfur substances (VSS). Nine components were identified on the basis of their mass spectra, namely, dimethyl disulfide, dimethyl trisulfide, diallyl disulfide, dipropyl disulfide, allyl methyl sulfide, allyl methyl trisulfide, 2-vinyl-4H-1,3-dithiin, 3,4-dihydro-3-vinyl-1,2-dithiin, and allithiamine. Extract DL-1 had the highest VSS content. Dried plant materials contained larger amounts of the VSSs than other extracts, and the leaves contained larger amount than the bulbs. These results suggest that heat treatment increases the antiheaptotoxic ability of A. victorialis var. platyphyllum by increasing the proportion of VSSs.

Safening Mode of Action of 1, 8-Naphthalic Anhydride on Corn and Soybean Against Herbicide Bensulfuron and Imazaquin (제초제(除草劑) bensulfuron과 imazaquin에 대한 1, 8-naphthalic anhydride(NA)의 옥수수와 콩에 대한 약해경감작용기구(藥害輕減作用機構))

  • Hwang, I.T.;Choi, J.S.;Kim, J.S.;Cho, K.Y.;Chun, J.C.
    • Korean Journal of Weed Science
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    • v.14 no.1
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    • pp.8-15
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    • 1994
  • The mode of safening action and potency of the 1, 8-naphthalic anhydride(NA) were investigated in corn(Zea mays) and soybean(Glycine max) treated with herbicide bensulfuron[2-{{{{{(4,6-dimethoxy-2-pyrimidinyl)amino}carbonyl}amino}sulfonyl}methyl}benzoic acid] and imazaquin[2-{4,5-dihydro-4-methyl-4-(1-methylethyl)-5-oxo-1H-imidazol-2-yl} 3-quinolinonecarboxylic acid]. Seed dressing with 0.2%(w/w) NA showed successful protection in corn against injury from herbicide bensulfuron and imazaquin but not in soybean. Safening factors of NA against bensulfuron and imazaquin were 10.2 and 5.0, respectively, in corn, while they were both 1.3 in soybean. In vivo, Glutathione-S-transferase(GST) activity of NA-treated corn and soybean increased 1.8-and 1.3-fold, respectively, but the activity was not affected by the herbicides in vitro. Acetolactate synthase(ALS) levels of NA-treated corn was increased 1.3-fold, but not changed in soybean. Tolerance of ALS activity to the herbicides was slightly greater in ALS obtained from NA-treated corn than that from the untreated, whereas the difference was not found in soybean. A significant increase of ACCase due to NA occurred in corn, but not in soybean. The herbicides did not affect in vitro ACCase activity.

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In vivo Metabolism of Endosulfan in Carp (Cyprinus carpio) (In vivo 시험에 의한 잉어체내 $^{14}C-endosulfan$의 대사)

  • Lee, K.B.;Shim, J.H.;Suh, Y.T.
    • Applied Biological Chemistry
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    • v.37 no.3
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    • pp.203-209
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    • 1994
  • When $^{14}C-{\alpha}-endosulfan$ was incubated with carp liver, kidney and gut preparations, it was metabolized to water soluble and organosoluble metabolites. In an in vitro test, endosulfan was converted to endosulfan ${\alpha}-hydroxyether$ (EHE), endosulfan alcohol (EA) and endosulfan ether (EE). The addition of NADPH resulted in rapid conversion of endosulfan to the metabolites in 105,000 g soluble fraction and microsomes. However, the rate of metabolism of endosulfan in liver, kidney and gut supplemented with NADPH as a cofactor was higher in the 105,000 g soluble fraction than that in the microsomes of carp under incubation conditions. The enzymes probably involved in the metabolism of endosulfan include the glutathione S-transferase (GST) and the mixed function oxidases (MFO), based on the evidence that addition of either GSH or NADPH increased the degradation of endosulfan.

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Protective Effects of Water Extracts Composed of Adenophora triphylla var. japonica Hara on the Acetaminophen-induced Hepatotoxicity (아세트아미노펜에 의해 유도된 간독성 모델에서 잔대를 주원료로 하는 추출물의 간 보호 효과)

  • Gum, Sang-Il;Lee, Dong-Ung;Cho, Min-Kyung
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.688-693
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    • 2007
  • The hepatoprotective effects of water extracts composed of Adenophora triphylla var japonica Hara (ATJH) on acetaminophen (APAP)-induced hepatotoxicity were investigated in vivo and in vivo. The effects of ATJH on liver toxicity induced by APAP were assessed by blood biochemical and histopathological analyses. APAP treatment (350 mg/kg) caused severe liver injury in mice as indicated by their significantly elevated plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. Pretreatment with ATJH for 3 or 7 days attenuated the increases in ALT and AST when challenged with APAP. The reductions in viability caused by high dose of APAP (450 mg/kg) in vivo were reversed by pretreatment with ATJH. These protective effects of ATJH against APAP-induced toxicity were consistent with the results from the histopathological examinations. We next examined the effects of ATJH on the gene expression of glutathione S-transferases (GSTs) that detoxify the metabolic intermediates of APAP in H4IIE cells. The hepatic GST protein levels [$\alpha$ class (GSTA2, GSTA3/5)] were significantly elevated in a dose-dependent manner by ATJH treatment. In summary, ATJH is effective at protecting against APAP-induced hepatotoxicity by GST induction, implying that ATJH should be considered a potential chemopreventive agent.

Allopurinol Decreases Liver Damage Induced by Dermal Scald Burn Injury (피부 화상으로 유도된 간 손상에서 Allopurinol의 효과)

  • Cho, Hyun-Gug;Yoon, Chong-Guk;Park, Won-Hark
    • Applied Microscopy
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    • v.31 no.1
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    • pp.37-47
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    • 2001
  • In order to investigate a pathogenesis of liver damage induced by skin burn, thermal injury was induced by scald burn on entirely dorsal surface in rats (total burn surface area $20\sim25\%$) except for inhalated injury. At 5 and 24 h after scald burn, biochemical assay and morphological changes in serum and liver tissue were examined. Skin burn increased liver weight (% of body weight, p<0.05) and the activity of serum aniline amino-transferase (ALT, p<0.05), in addition, the activity of xanthine oxidase (XO), an enzyme of oxygen free radical generating system, was elevated (p<0.01) in serum, but not in skin and in liver. Postburn treatment of allopurinol intraperitoneally decreased liver weight, serum ALT activity and serum XO activity. Scald burn induced ultrastructurally swelling of endoplasmic reticulum, ribosome detachment, accumulation of lipid, dilatation of bile canaliculi and intercellular space, neutrophil infiltration, activation of Kupffer's cells and degeneration of hepatocytic microvilli. Futhermore , thermal injury decreased not only the protein concentration in plasma but also the number of intravascular leukocytes, that indicates induction of edema formation with protein exudation and inflammation by neutrophil infiltration into the internal organs. However allopurinol injection after burn inhibited post burn ultrastructural changes. These data suggest that acute dermal scald burn injury leads to liver damage, that is related to elevation of xanthine oxidase activity in serum. Xanthine oxidase may be a key role in the pathogenesis of liver damage induced by skin burn.

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Development of mixed Th1/Th2-type immune response in mice following immunization with GP63 from Leishmania donovani (내장리슈만편모충 유래 GP63 항원을 마우스에 접종한 후 관찰되는 Th1/Th2-type 복합 면역반응)

  • Shin, Sung-shik
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.211-218
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    • 2001
  • The $M_r$ 63,000 glycoprotein (GP63) and lipophosphoglycan (LPG) of Leishmania donovani were evaluated as vaccine candidates against visceral leishmaniasis. Mice were immunized with liposomeencapsulated GP63 and/or LPG that were purified from the soluble extract of L. donovani promastigotes, and were challenged with virulent amastigotes. Mice immunized with GP63/LPG in liposomes plus BCG resulted in a 27.4% reduction of amastigotes in the liver compared to the control group (liposomes plus BCG), and mice immunized with liposome-GP63 plus BCG failed to induce a protective immune response against the challenge infection. Immunization of mice with GP63 fused to the Schistosoma japonicum glutathione S-transferase (GP63-GST) plus BCG also failed to elicit protective immunity. To analyze the cause of failure to induce protection, cytokine release from the spleen cells of immunized mice and Leishmania-specific serum antibodies were analyzed. Spleen cells from mice immunized with GP63-GST plus BCG that were exposed to soluble extract of L. donovani in vitro produced 10-fold greater quantities of IFN-gamma and 3-fold greater quantities of IL-5 than cells from mice receiving BCG only or saline. Western blot analysis revealed that sera from these mice had Leishmania-specific antibodies recognizing 1 to 3 antigens of L. donovani with M. W. of 60-65 kDa. Although immunization of mice with GP63-GST induced a strong Th1 response, this study indicated that GP63-GST simultaneously elicited the Th2 response of the CD4+ T-cell, which was known to abrogate the protective immune response conferred by the Th1 effector function.

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Water Extract of Rosa laevigata Michx. Protects Hepatocytes from Arachidonic Acid and Iron-mediated Oxidative Stress (아라키돈산과 철 유도성 산화적 스트레스에 대한 금앵자(金櫻子) 열수 추출물의 간세포 보호 효능)

  • Ko, Hae Li;Jegal, Kyung Hwan;Song, Si Yeon;Kim, Nan Ee;Kang, Jiwon;Byun, Sung Hui;Kim, Young Woo;Cho, Il Je;Kim, Sang Chan
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.7-15
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    • 2015
  • Objectives : Rosa laevigata Michx. has been used for the treatment of renal disease in traditional Korean medicine. In this study, we investigated cytoprotective effect of R. laevigata water extract (RLE) against oxidative stress induced by arachidonic acid (AA) + iron.Methods : To evaluate the protective effects of RLE against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The effects of RLE on reduced glutathione level, production of reactive oxygen species and mitochondrial membrane potential were also monitored. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) was examined by immunoblot analysis. Additionally, Nrf2 transactivation and its downstream target genes expression were also determined by reporter gene and realtime RT-PCR analyses.Results : RLE pretreatment (30-300 μg/ml) prevented cells from AA + iron-mediated cell death in a concentration dependent manner. In addition, 100 μg/ml RLE inhibited AA + iron-induced glutathione depletion, reactive oxygen species production and mitochondrial dysfunction. RLE accumulated nuclear Nrf2 and also transactivated Nrf2, which was evidenced by antioxidant response element- and glutathione S-transferase A2-driven luciferase activities and mRNA level of glutamate-cysteine ligase catalytic subunit, NAD(P)H:quinone oxidoreductase 1 and sestrin 2. Moreover, protective effect of RLE against AA + iron was abolished in Nrf2 knockout cells.Conclusions : These results indicate that RLE has the ability to protect hepatocyte against oxidative stress through Nrf2 activation.