• Title/Summary/Keyword: Oxidase

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A Molecular Study of Sopungsungi-won(Shufengshunqiyuan) about Regulation of PPARs in Mouse NMu2Li Liver Cells and C2C12 Skeletal Muscle Myogenic Progenital Cells (소풍순기원(疏風順氣元)이 mouse의 NMu2Li 간세포와 C2C12 골격근세포에서 PPARs 조절의 분자기전에 미치는 영향)

  • Oh, Young-Jin;Shin, Soon-Shik;Yoon, Mi-Chung;Kim, Bo-Kyung
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.1
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    • pp.147-164
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    • 2009
  • Objectives : We investigated the effects of Sopungsungi-won(Shu!engshunqiyuan) (SSEx1, SSEx2) to treat the metabolic syndrome by the molecular mechanism of regulation of PPAR and modulation of mitochondrial MCAD, VLCAD mRNA expression. Methods : Mouse NMu2Li liver cells and C2C12 skeletal muscle myogenic progenital cells were transiently transfected with expression plasmids for PPAR(PPAR${\alpha}$, PPAR${\delta}$), a luciferase reporter gene construct containing 3 copies of the PPRE from the rat acyl-CoA oxidase gene and ${\beta}$-galactosidase gene. Cells were treated with several concentrated kinds of SSEx1, SSEx2 at the initial time of culture and analyzed PPAR${\alpha}$, PPAR${\delta}$ reporter gene activity using spectrophotometer (405 nm). Total RNA was extracted from SSEx1, SSEx2 and measured mRNA levels of mitochondrial MCAD, VLCAD. Representative RT-PCR bands are shown. Results : 1. SSEx1 increased the expression of PPAR${\alpha}$ reporter gene activities at 0.1 ${\mu}$g/ml (p${\mu}$g/ml (p<0.05), SSEx2 at 0.1 ${\mu}$g/ml (p${\mu}$g/ml (p<0.05) significantly in NMu2Li liver cell lines. 2. SSEx1 increased the expression of PPAR${\alpha}$ reporter gene activities at 1 ${\mu}$g/ml (p${\mu}$g/ml (p${\alpha}$ reporter gene activities in C2C12 skeletal muscle cells. 4. SSEx1 increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in NMu2Li liver cell lines. 5. SSEx1, SSEx2 both increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in C2C12 skeletal muscle cells. Conclusions : These results show the SSEx1, SSEx2 can be used as therapeutic agent for metabolic syndrome and it's molecular mechanisms of PPAR more contribute to the activation of PPAR${\alpha}$ then PPAR${\delta}$ reporter gene activities and it's total RNA more contribute to the modulation of mitochondrial MCAD then VLCAD mRNA expression.

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Effect of Bambusae caulis in Liquamen on Streptozocin-Induced Diabetic C57BL/6 Mice

  • Deung Young-Kun;Park Seung-Kyu;Jin Dan;Yang Eun-Ju;Lim Soo-Jung;Kwon Ki-Rok;Kim Dong-Heui;Lee Kyu-Jae
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.343-347
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    • 2005
  • Bambusae caulis in Liquamen is one of the important herbal medicine produced by heating bamboo indirectly and is used for treatment of stroke, hypertension, and diabetes etc. Recently the mechanism of clinical effects on Bambusae caulis in Liquamen has been studied. This experiment was conducted to confirm the clinical effects of Bambusae caulis in Liquamen on type 1 diabetes and its related mechanism. We divided C57BL/6 mice into 3 groups and induced them to be type 1 diabetes by injection of streptozocin into peritoneum. The dosage of each group was 150 mg/kg once only, 140 mg/kg once only and 40 mg/kg for 5 days respectively. The two groups injected streptozocin for once took orally Bambusae caulis in Liquamen after the induction of diabetes, and the other one group was given Bambusae caulis in Liquamen during the diabetes inducing period. As the result, the two diabetes-induced groups showed blood glucose decreasing effect by Bambusae caulis in Liquamen on an average, but they didn't show the signiftcant differences statistically. But Bambusae caulis in Liquamen showed the anti-diabetic effect suppressing blood sugar rising trend during the diabetes inducing peried (P<0.05). The anti-oxidative effect of Bambusae caulis in Liquamen was measured with the hypoxanthine/xanthine oxidase (HX/XOD) system. The quantity of ROS was measured using DCFDA reagent indirectly. As the result, $10\%$ solution of Bambusae caulis in Liquamen showed anti-oxidative effect by scavenging $93.4\%$ superoxide as compared with control group. It is suspected that the anti-oxidative effect of Bambusae caulis in Liquamen suppressed the increase of blood glucose in the diabetes-inducing group. These results could be useful data to understand the effect of Bambusae caulis in Liquamen on type 1 diabetes and type 1 diabetes developing because ROS were closely connected with the induction and complications of diabetes.

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Effects of Solvent-extracted Fractions from Salicornia herbacea on Anti-oxidative Activity and Lipopolysaccharide-induced NO Production in Murine Macrophage RAW264.7 Cells

  • Lee, Whi-Min;Sung, Hye-Jin;Song, Jae-Chan;Cho, Jae-Youl;Park, Hwa-Jin;Kim, Suk;Rhee, Man-Hee
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.161-168
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    • 2007
  • Salicornia herbacea L. (Chenopodiaceae: S. herbacea) is a salt marsh plant, which has long been prescribed in traditional medicines for the treatment of intestinal ailments, nephropathy, and hepatitis in Oriental countries. In order to elucidate the mechanisms of this herb, we conducted an anti-oxidative activity, the inhibition of nitric oxide (NO) production, and the suppression of the pro-inflammatory cytokine genes, with the solvent-extracts of S. herbacea. We found that both ethyl acetate and n-butanol tractions showed potent anti-oxidative effects in comparison to other fractions using xanthine oxidase assay with $IC_{50}$ values of $66.0{\pm}0.5\;{\mu}g/ml$ and $82.5{\pm}3.8\;{\mu}g/ml$, respectively. In addition, both ethyl acetate and n-butanol fractions showed more electron donating activity (EDA) than other tractions, according to DPPH (2, 2-Diphenyl-lpicrylhydrazyl radical) assay. The EDA of ethyl acetate fraction ($IC_{50}$ values of $117.5{\pm}3.8\;{\mu}g/ml$) is more significant than that of n-butanol fraction ($IC_{50}$ values of $375.0{\pm}12.5\;{\mu}g/ml$). Among potential anti-oxidative tractions, ethyl acetate traction dose-dependently suppressed lipopolysaccharide (LPS, $0.1\;{\mu}g/ml$)-induced nitric oxide (NO) production in RAW264.7 cell, while n-butanol did not. As expected, ethyl acetate fraction suppressed the expression of inducible NO synthase (iNOS) in RAW264.7 cell stimulated by $0.1\;{\mu}g/ml$ of LPS. Moreover, the ethyl acetate traction suppressed the expression of interleukin-1 $(IL)-1{\beta}$ and granulocyte/macrophage colony-stimulating factor (GM-CSF) mRNA in LPS-stimulated RAW264.7 cells. Therefore, these results suggest that S. herbacea may have anti-oxidative and anti-inflammatory activities by modulating radical-induced toxicity and various pro-inflammatory responses.

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Effect of Dried Powders and Water Extracts of Paecilomyces Tenuipes and Cordyceps Militaris on Lipid Metabolism, Antioxidative Capacity and Immune Status in Rats (동충하초의 건분 및 물추출물이 흰쥐의 지방대사, 항산화 및 면역능에 미치는 효과)

  • Kwon, Sang-Hee;Woo, Hee-Jong;Han, Dae-Seok;Kim, Mi-Kyung
    • Journal of Nutrition and Health
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    • v.34 no.3
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    • pp.271-284
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    • 2001
  • This study was performed to investigate the effects of dried powders and water extracts of Paecilomyces tenuipes(P. tenuipes) and Cordyceps militaris(C. militaris) on lipid metabolism, lipid peroxidation and antioxidative capacity and immune status in rats. Thirty-five male Sprague-Dawley rats weighting 195$\pm$21g were grouped into five according to body weight. Ratw were raised for four weeks with diet containing either 4%, 2%(w/w) of dried P. tenuipes powders(TP-4, TP-2) or water extracts from equal amounts of each 4% P. tenuipes and C. militaris powder(TE-4, ME-4). Food intake, weight gain of all groups were not significantly different from those of control group. Lipid metabolism in general was not significantly different among all the groups. However both dried P. tenuipes powder lowered plasma cholesterol level slightly, water extract groups showed tendency of higher plasma HDL-cholesterol and lower liver cholesterol levels than control. Plasma and liver thiobarbituric acid reactive substance(TBARS) concentrations of all the experimental groups were lower than control group. Red blood cell(RBC) and liver superoxide dismutase(SOD) activities were not generally different among all groups. Liver xanthine oxidase(XOD) activities of all groups were tended to be lower than control group. Proliferation of aplenocytes induced by mitogens, concanavalin A and lipopolysaccharide, were increased in TP-2 group. The TP-4 group showed increased CD8 T cells and MHC class II expression without changes in CD4 T cells, B cells and G/M ratio, suggesting activated cytotoxic T cell activity in vivo. Increase of G/M ratio but not of MHC class II in TP-2 group indicated the possible acute inflammatory reaction by the ingested substances in gastrointestinal tract. ME-4 group showed enhanced cellular immunity without vigorous changes of immune parameters in brief periods. In conclusion, both P. tenuipes and C. militaris stimulated antioxidant capacity and immune status in rats. Among groups, water extract of C. militaris was most effective in both capacities, though dried powder of P. tenuipes at 2% dietary level was more effective in antioxidant activity, as various results by different strains were observed.(Korean J Nutrition 34(3) : 271~284, 2001)

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Effect of Methionine Levels on Brain Lipid Peroxidation in Ethanol-treated Rats of Selenium Deficiency (메티오닌과 셀렌이 에탄올 중독된 흰쥐의 뇌지질과산화에 미치는 영향)

  • 조수열;이미경;박은미;장주연;김명주
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.109-115
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    • 1997
  • This study was designed to investigate the effects of methionine(Met) on the activities of brain lipid peroxidation related enzymes in ethanol administrated rats of selenium(Se) deficiency. Male Sprague-Dawley rats were fed Se deficiency diets containing one of the three levels of Met (0, 3, 9g/kg diet) and ethanol(2.5g/kg of body weight) was administrated as 25v/v% ethanol treated groups orally. The rats sacrificed after 5 and 10 weeks of feeding periods. Alcohol dehydrogenase activity was increased in ethanol treated groups and was higher Met normal group than Met deficiency and excessive groups at 5 and 10 weeks dieting. Aldehyde dehydrogenase activity was decreased in ethanol treated groups and significantly decreased in Met deficiency group. Monoamine oxidase activity in brain was increased in ethanol treated groups and was predominently increased in Met deficiency groups. Superoxide dismutase and glutathione peroxidase activities were decreased in ethanol treated groups and tended to increase in proportion to level of dietary methionine. Glutathione S-transferase and catalase activities and lipid peroxide content were increased by ethanol administration and were higher Met deficiency group than normal and excessive groups.

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Comparative analysis on genome-wide DNA methylation in longissimus dorsi muscle between Small Tailed Han and Dorper×Small Tailed Han crossbred sheep

  • Cao, Yang;Jin, Hai-Guo;Ma, Hui-Hai;Zhao, Zhi-Hui
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1529-1539
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    • 2017
  • Objective: The objective of this study was to compare the DNA methylation profile in the longissimus dorsi muscle between Small Tailed Han and Dorper${\times}$Small Tailed Han crossbred sheep which were known to exhibit significant difference in meat-production. Methods: Six samples (three in each group) were subjected to the methylated DNA immunoprecipitation sequencing (MeDIP-seq) and subsequent bioinformatics analyses to detect differentially methylated regions (DMRs) between the two groups. Results: 23.08 Gb clean data from six samples were generated and 808 DMRs were identified in gene body or their neighboring up/downstream regions. Compared with Small Tailed Han sheep, we observed a tendency toward a global loss of DNA methylation in these DMRs in the crossbred group. Gene ontology enrichment analysis found several gene sets which were hypomethylated in gene-body region, including nucleoside binding, motor activity, phospholipid binding and cell junction. Numerous genes were found to be differentially methylated between the two groups with several genes significantly differentially methylated, including transforming growth factor beta 3 (TGFB3), acyl-CoA synthetase long chain family member 1 (ACSL1), ryanodine receptor 1 (RYR1), acyl-CoA oxidase 2 (ACOX2), peroxisome proliferator activated receptor-gamma2 (PPARG2), netrin 1 (NTN1), ras and rab interactor 2 (RIN2), microtubule associated protein RP/EB family member 1 (MAPRE1), ADAM metallopeptidase with thrombospondin type 1 motif 2 (ADAMTS2), myomesin 1 (MYOM1), zinc finger, DHHC type containing 13 (ZDHHC13), and SH3 and PX domains 2B (SH3PXD2B). The real-time quantitative polymerase chain reaction validation showed that the 12 genes are differentially expressed between the two groups. Conclusion: In the current study, a tendency to a global loss of DNA methylation in these DMRs in the crossbred group was found. Twelve genes, TGFB3, ACSL1, RYR1, ACOX2, PPARG2, NTN1, RIN2, MAPRE1, ADAMTS2, MYOM1, ZDHHC13, and SH3PXD2B, were found to be differentially methylated between the two groups by gene ontology enrichment analysis. There are differences in the expression of 12 genes, of which ACSL1, RIN2, and ADAMTS2 have a negative correlation with methylation levels and the data suggest that DNA methylation levels in DMRs of the 3 genes may have an influence on the expression. These results will serve as a valuable resource for DNA methylation investigations on screening candidate genes which might be related to meat production in sheep.

Effect of Water and Ethanol Extracts of Persimmon Leaf and Green Tea Different Conditions on Lipid Metabolism and Antioxidative Capacity in 12-month-old Rats (추출 조건을 달리한 감잎과 녹차의 물 및 에탄올 추출물이 노령쥐의 지방대사와 항산화능에 미치는 영향)

  • 김성경;이혜진;김미경
    • Journal of Nutrition and Health
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    • v.34 no.5
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    • pp.499-512
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    • 2001
  • This study was performed to investigate effects of dried leaf powders, water, 75% and 95% ethanol extracts of persimmon leaf and green tea on lipid metabolism, lipid peroxidation and antioxidative enzyme activity in 12-month-old rats. Fifty-four male Sprague-Dawley rats weighing 542$\pm$4.5g were blocked into groups according to their body weight and were raised for four weeks with the diets containing 5%(w/w) dried leaf powders of persimmon(Diospyros kaki Thunb) and green tea(Camellia Sinensis O. Ktze), water or 75% and 95% ethanol extracts from same amount of each dried tea powder. Food intake was not significantly different among all groups, but weight gain of green tea powder group was significantly lower than that of control group. Plasma and liver lipid levels of all the tea diet groups were lower than those of control group. Especially, 75% ethanol extract of persimmon leaf decreased total lipid and triglyceride concentrations in plasma and 95% ethanol extract of persimmon leaf decreased liver total lipid level. However, there was no difference between 75% ethanol extracts groups and 95% ethanol extracts groups in lipid metabolism. Superoxide dismutase(SOD) and catalase activities in erythrocyte were remarkably increased by all the green tea diets. SOD, catalase and glutathione peroxidase activities in liver were increased by the feeding of ethanol extracts from green tea and persimmon leaf powder. Liver xanthine oxidase activity was not different among all groups. Plasma Thiobarbirutic acid reactive substance(TBARS) concentrations of all the green tea diet groups were significantly low. It was thought that high flavonoids in green tea inhibited plasma lipid peroxidation by promoting SOD, catalase activities in erythrocyte. 95% ethanol extract of persimmon leaf also inhibited plasma lipid peroxidation by high vitamin E and beta-carotene. Persimmon leaf powder decreased liver TBARS concentration by vitamin E, betacarotene and vitamin C and by increasing activities of antioxidative enzymes with flavonoids. In conclusion, dried leaf powders, water, 75% and 95% ethanol extracts of persimmon leaf and green tea were effective in lowering lipid levels and inhibiting lipid peroxidation in 12-month-old rats. Above all, ethanol extracts of persimmon leaf decreased plasma and liver lipid levels and persimmon leaf powder effectively inhibited liver lipid peroxidation. Extracts of green tea leaf inhibited plasma lipid peroxidation. In lowering lipid levels and inhibiting lipid peroxidation, ethanol extracts were more effective than water extracts, but there was no difference between 75% ethanol extracts and 95% ethanol extracts in lipid metabolism. (Korean J Nutrition 34(5) : 499~512, 2001)

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Effect of Dry Powders, Ethanol Extracts and Juices of Radish and Onion on Lipid Metabolism and Antioxidative Capacity in Rats (무, 양파의 시료제조 방법에 따른 흰쥐의 지방대사와 항산화능에 관한 연구)

  • 안소진;김미경
    • Journal of Nutrition and Health
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    • v.34 no.5
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    • pp.513-524
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    • 2001
  • This study was performed to investigate the effects of dry powders, ethanol extracts and juices of radish and onion on lipid metabolism, lipid peroxidation and antioxidative enzyme activity in rats. Forty-nine male Sprague-Dawley rats weighing 157$\pm$6g were blocked into seven groups according to body weight and raised for four weeks with diets containing 5%(w/w) dry powders of two different vegetables consumed frequently by Korean-radish(Raphanus sativus L.) and onion(Allium cepa L.), ethanol extracts and juices from equal amount of each dry powder. All the powders, ethanol extracts and juices of radish and onion decreased total lipids, triglycerides and total cholesterol concentrations in plasma and liver. Above all, onion ethanol extract decreased them most remarkably. It was thought that organosulfur compounds and flavonoids extracted from onion by ethanol inhibited biosynthesis and absorption of lipid and promoted degradation of lipid. Radish powder also decreased them by increasing fecal excretions of total lipids, triglycerides and total cholesterol most effectively. Catalase and glutathine peroxidase(GSH-px) activities in red blood cell(RBC) were most remarkably increased by radish powder and onion powder respectively. Superoxide dismutase(SOD), catalase and GSH-px activities in liver were most remarkably increased by onion ethanol extract, radish powder and onion ethanol extract respectively. Xanthine oxidase(XOD) activities in liver were most effectively decreased by ethanol extracts of radish and onion. Thiobarbituric acid reactive substance (TBARS) levels in plasma and liver of experimental groups were significantly lower than those of controls. Above all, onion powder decreased them most effectively. It was thought that vitamin E and high flavonoids in onion powder inhibited lipid peroxidation, promoting liver and RBC SOD, catalase and GSH-px activities and inhibiting XOD activities effectively. Flavonoids in onion ethanol extract inhibited lipid peroxidation, promoting three antioxident enzyme activities and inhibiting XOD activities most remarkably. Also flavonoids and high vitamin C in radish powder inhibited lipid peroxidation, promoting liver and RBC catalase most remarkably and inhibiting XOD activities. In conclusion, radish and onion were effective in lowering lipid levels and inhibiting of lipid peroxidation in animal tissue. From these data, radish and onion can be recommended in the treatment and prevention of diseases such as cardiovascular disease and cancer and in delaying aging. As ethanol from onion were most effective in lowering lipid level and promoting three antioxident enzymes, and inhibited lipid peroxidation as did we should try to utilize onion skin which is discarded though reported to have abundant flavonoids. (Korean J Nutrition 34(5) : 513~524, 2001)

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Effect of Dried Powders or Ethanol Extracts of Garlic Flesh and Peel on Antioxidative Capacity in 16-month-old Rats (마늘 육질과 껍질의 건분 및 에탄올추출물이 노령흰쥐의 항산화능에 미치는 영향)

  • 신성희;김미경
    • Journal of Nutrition and Health
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    • v.37 no.8
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    • pp.633-644
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    • 2004
  • This study was performed to investigate effect of dried powders and ethanol extracts of garlic flesh and peel on antioxidative capacity in 16-month- old rats. Forty Sprague-Dawley male rats weighing 618.1$\pm$6.5g were blocked into five groups according to body weight and raised for 3 months with experimental diets containing 5% (w/w) of dried powders of garlic flesh or peel, or ethanol extracts from equal amount of each dried powder. Total polyphenols, flavonoids, /3 -carotene, vitamin C, vitamin E, and total antioxidant status (TAS) levels were determined in garlic preparations. Thiobarbituric acid reactive substances (TBARS) levels in plasma, liver and VLDL + LDL fraction, oxidative DNA damage (8-hydroxy-2' -deoxyguanosine, 80HdG) in kidney, xanthine oxidase (XO) activities in plasma and liver, superoxide dismutase (SOD) activities in erythrocyte and liver, and carotenoid concentration, and total antioxidant status (TAS) in plasma were measured. Total polyphenols and flavonoids contents in garlic preparations were highest in peel ethanol extract. Vitamin C content was not different significantly among preparations, but peel powder contains slightly more vitamin C. The content of $\beta$-carotene was highest in peel ethanol extract and vitmain E content was highest in flesh ethanol extract. The highest level of TAS was observed in peel ethanol extract. Plasma TBARS levels in all the experimental groups were found to be significantly lower than control group, and TBARS concentration in VLDL + LDL fraction was decreased in all the experimental groups in comparison to control group. Also levels of 80HdG in kidney in experimental groups were lower than that of control group. Plasma and liver XO activities were. decreased in all experimental groups, and erythrocyte and liver SOD activities were higher in experimental groups compared to control group. All experimental groups also showed higher plasma TAS levels than control group. Especially, garlic flesh powder group was significantly lower in plasma and liver XO activities, and significantly higher in erythrocyte and liver SOD activities than control group. Moreover, plasma TBARS level and kidney 8OHdG level were decreased in flesh powder group. In conclusion, garlic diets showed effect of improving antioxidative capacity in 16-month old rats, especially, garlic flesh powder was prominent in inhibiting XO activity, promoting SOD activity and decreasing kidney 8OHdG level among experimental groups.

Human brain pyridoxal-5'-phosphate phosphatase (PLPP): protein transduction of PEP-1-PLPP into PC12 cells

  • Lee, Yeom-Pyo;Kim, Dae-Won;Lee, Min-Jung;Jeong, Min-Seop;Kim, So-Young;Lee, Sun-Hwa;Jang, Sang-Ho;Park, Jin-Seu;Kang, Tae-Cheon;Won, Moo-Ho;Cho, Sung-Woo;Kwon, Oh-Shin;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.41 no.5
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    • pp.408-413
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    • 2008
  • Pyridoxal-5'-phosphate phosphatase (PLPP) catalyzes the dephosphorylation of pyridoxal-5'-phosphate (PLP). A human brain PLPP gene was fused with a PEP-1 peptide and produced a genetic in-frame PEP-1-PLPP fusion protein. The purified PEP-1-PLPP fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced PEP-1-PLPP fusion protein was stable for 36 h. The concentration of PLP was markedly decreased by the addition of exogenous PEP-1-PLPP to media pretreated with the vitamin $B_6$ precursors; pyridoxine, pyridoxal kinase and pyridoxine-5'-phosphate oxidase into cells. The results suggest that the transduction of the PEP-1-PLPP fusion protein can be one mode of PLP level regulation, and to replenish this enzyme in the various neurological disorders related to vitamin $B_6$.