The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.
Dietary effect of soyfiber beni-koji (SBK) with chitosan-ascorbate (CA) on the level of serum lipids in rats fed a high fat diet was investigated The experimental groups which were divided into high fat diet control group(HC-group), $2\%\;SBK+0.1\%$ CA mixture diet group(CA1-group), $2\%\;SBK+0.15\%$ CA mixture diet group(CA2-group), and $2\%\;SBK+0.2\%$ CA mixture diet group (CA2-group) were fed for 4 weeks. Weight gains in CA2- and CA3-group were $5.3\%\;and\;9.5\%$ lower than that of HC-group, respectively, while there was no significant difference in feed intakes, feed efficiency ratio and organs weight Level of serum triglyceride in $C_3-group\;was\;21\%$ lower than that of HC-group. Level of serum total cholesterol and LDL-cholesterol in CA2- and CA3-group were $22.1\~22.7\%\;and\;58.6\~64.3\%$ lower than those of HC-group, respectively. Atherogenic index decreased with the higher level of CA. Level of lipid peroxide in CA3-group was $24\%$ lower than that of HC-group, while there was no significant difference in GSH(Glutathione-S-transferase) content O type activities of XOD(xanthine oxidase) in the treated groups were lower, especially the activity in CA3-group was $51.6\%$ lower than that of HC-group. Also, O/T ratio of XOD was lower, showing $21.7\~23.5\%$ in treated groups and $34.0\%$ in HC-group(p<0.05). GST activities were 332.52 units in HC-group and $350.28\~355.63$ units in the treated groups, but there were no significant differences among them.
Journal of the Korean Society of Food Science and Nutrition
/
v.30
no.5
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pp.928-932
/
2001
The protective effect of Hericium erinaceus methanol extract was investigated on benzo($\alpha$)pyrene(B($\alpha$)P) induced hepatotoxicity in mice, Hericium erinaceus extract was intraperitioneally injected once a hay for successive 5 days. followed by treatment with B($\alpha$)P on the fifth day. The elevated activities of serum aminotransferase and hepatic cytochrome P-450 by B($\alpha$)P was decreased by pretretament with Hericium erinaceus extract. Moreover, hepatic lipid peroxide content and glutathions S-transferase activity increased by B($\alpha$)P were significantly decrease, but depletion of glutathione content induced by B($\alpha$)P was prevented by Hericium erinaceus extract. In addition, the increased activities of superoxide diamutase, catalase and glutathions peroxidase after B($\alpha$)P-treatment were decreasd. Immunoblott analysis of hepatic microsomes showed that methanol extract of Hericium erinaceus suppressed protein level of the cytochrome P-450 1AI increaed by B($\alpha$)P. These results suggest that Hericium erinaceus extract may protect liver from damage induced by B($\alpha$)P.
Zhengxuan, Wang;Mingcai, Liang;Hui, Li;Bingxiao, Liu;Lin, Yang
Nutrition Research and Practice
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v.16
no.6
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pp.729-744
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2022
BACKGROUND/OBJECTIVES: 4-Hydroxy-2-nonenal (HNE) is a biomarker for oxidative stress to induce inflammation. Methionine is an essential sulfur-containing amino acid with antioxidative activity. On the other hand, the evidence on whether and how methionine can depress HNE-derived inflammation is lacking. In particular, the link between the regulation of the nuclear factor-κB (NF-κB) signaling pathway and methionine intake is unclear. This study examined the link between depression from HNE accumulation and the anti-inflammatory function of ⳑ-methionine in rats. MATERIALS/METHODS: Male Wistar rats (3-week-old, weighing 70-80 g) were administered different levels of ⳑ-methionine orally at 215.0, 268.8, 322.5, and 430.0 mg/kg body weight for two weeks. The control group was fed commercial pellets. The hepatic HNE contents and the protein expression and mRNA levels of the inflammatory mediators were measured. The interleukin-10 (IL-10) and glutathione S-transferase (GST) levels were also estimated. RESULTS: Compared to the control group, hepatic HNE levels were reduced significantly in all groups fed ⳑ-methionine, which were attributed to the stimulation of GST by ⳑ-methionine. With decreasing HNE levels, ⳑ-methionine inhibited the activation of NF-κB by up-regulating inhibitory κBα and depressing phosphoinositide 3 kinase/protein kinase B. The mRNA levels of the inflammatory mediators (cyclooxygenase-2, interleukin-1β, interleukin-6, inducible nitric oxide synthase, tumor necrotic factor alpha) were decreased significantly by ⳑ-methionine. In contrast, the protein expression of these inflammatory mediators was effectively down regulated by ⳑ-methionine. The anti-inflammatory action of ⳑ-methionine was also reflected by the up-regulation of IL-10. CONCLUSIONS: This study revealed a link between the inhibition of HNE accumulation and the depression of inflammation in growing rats, which was attributed to ⳑ-methionine availability. The anti-inflammatory mechanism exerted by ⳑ-methionine was to inhibit NF-κB activation and to up-regulate GST.
Screening of Biologically Active Essential Oils from Ligusticum tenuissimum. Kim, Min-Hae, Young-Gil Kim, Jin-Ha Lee, Keo-Pyo Hong, Jung-Ki Hong, Young-Joon Kong, and Hyeon-Yong Lee*. Division of Food and Biotechnology, Kangwon National University, Chunchon 200-701, Korea, 1 Regional Crop Development Station, Kangwon Agricultural Research & Extension Services, Chunchon 200-150, Korea-The biological activities of the crude essential oils from Ligusticum tenuissimum and the control(phthalic anhydride) were compared. About 60% of the growth of MCF7, A549, and Rep3B cells were inhibited by adding 1.0 mg/ml of the crude essential oils and below 40% was observed by the control. Cytotoxicity on human normal lung cell(IMR90) was scored as 34.4% for the crude oil and 26.4% for control, respectively. It was found that the crude essential oils were more effective than the control in anti mutagenecity tested by both Rec-assay and CRG V79 cells. The growth of human T-cell(Jurkat) was enhanced up to 1.21 times by adding the crude essential oil compared with the control. 50% of a-glucosidase activity was inhibited by both the crude essential oil and the control. ACE activities were inhibited 80.1 % and 65.3% by adding 1.0 mg/ml of the crude oil and the control, respectively. The higher enhancement of glutathione-S-transferase activity was observed in the crude oil than those in the control: 301 % v.s 234% at 1.0 mg/ml of the treatment. Thrombolytic activity was measured as 42.9% and 28.6% for the crude oil and the standard, respectively. The effect of the oil on the nerve cells PCI2, was observed as follows: the neurite of PCl2 cells was lengthened up to 255 /-lm longer than 205 /-lm of control. The number of neurite-bearing cells were about two times higher than control. The survival ratio of the crude essential oil was also increased up to 56.4% which was about two fold higher than in control.
The current study was performed to develop natural bio-active substances as additives for the production of high quality broiler chickens. A total of 120 male 3 day-old broiler chicks were randomly allocated to CON (control), GK2.5 (ginkgo leaf 2.5%), GK5.0 (ginkgo leaf 5.0%), PK2.5 (pumpkin 2.5%) and PK5.0 (pumpkin 5.0%) of five groups in cages (24 birds per group). All birds were fed corresponding diets from 3 to 35 d of age and determined growth performance and biological parameters including blood biochemical profiles, antioxidant status and intestinal microflora. During the entire feeding trial, GK5.0 and PK5.0 groups resulted in a significantly (P<0.05) higher FCR than GK2.5 and PK2.5 groups. Plasma triglyceride significantly (P<0.05) increased in GK5.0 group compared with the other groups, and the level of alanine transaminase (ALT) increased (P<0.05) in GK5.0 and PK5.0 groups compared with that in PK2.5 group. Dietary addition of ginkgo leaf and pumpkin significantly (P<0.05) increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the small intestine. Also, the addition of 2.5% ginkgo leaf significantly (P<0.05) increased the activities of SOD, GSH-Px and glutathione-S-transferase (GST) in the liver. Futhermore, muscle GST activity significantly (P<0.05) enhanced by dietary addition of ginko leaf and pumpkin. However, the level of lipid peroxidation (MDA) in the small intestine and muscle turned to be higher (P<0.05) in PK5.0 group. The colony forming units (CFU) of E. coli in intestinal digesta significantly (P<0.05) decreased in both ginko leaf and pumpkin supplemented groups compared with CON group. In conclusion, dietary addition of 2.5% ginko leaf and pumpkin as dietary sources can be applicable for the production of high quality broiler chickens.
This study was performed to investigate anti-thrombogenic, anti-inflammatory effects of n-BuOH (B) and $CH_2Cl_2$ (MC) fractions extracted from Sancho (Zanthoxylum. schinifolium) leaves in rats fed high fat diets. The experimental animal groups were consisted of eight including one 5% fat (N) and one 20% fat (H) without the test materials in diets and six H groups of feeding three levels (50, 100 and 150 mg/day) of the B and the MC fractions from Z. schinifolium, respectively. Plasma activated partial thromboplastin times and thrombin times of H group were decreased compared to the N group, but they were increased by feeding the MC fraction of 50 mg and over. Polymorphonuclear leukocyte 5#-lipo-xygenase activities and leukotriene $B_4$ contents of the H group were significantly increased compared to the N group, but they were decreased in the 100 mg and 150 mg of B fraction or the 150 mg of MC fraction fed groups. Liver cytochrome $P_{450}$, $O_2^-$, $H_2O_2$ and GSSG contents were increased by the high fat diet but decreased by feeding the B fraction or the MC fraction, while GSH content and glutathione S-transferase activity lowered by high fat diet were increased by feeding the two solvent fractions. The effects of the solvent fractions were evident at the level of 100 mg/day and over. The present results confirmed that two solvent fractions from the leaves of Z, schinifolium have enhancing effects on anti-thrombosis and anti-inflammation partly by antioxidant action and partly by direct modulation of the respective processeds. In conclusion, the n-BuOH and $CH_2Cl_2$ fractions from leaves of Z, schinifolium can be utilized as the proper ingredients of functional foods for preventing chronic degenerative disease.
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.7
/
pp.1043-1053
/
2013
We investigated the ability of soybean curd residue (SCR) and its fermented products to inhibit obesity and improve the blood lipid profiles of obese mice fed a high-fat diet. Samples were prepared by fermenting SCR with Aspergillus oryzae var effuses KACC 44990 (ASCR), a microbe used for the fermentation of traditional Korean Meju, and with Monascus pilosus IFO 4480 (MSCR), a microbe used for the production of red rice. In addition, AMSCR, a mixture composed of equal amounts of ASCR and MSCR, was also prepared. Male mice were divided into six groups and fed with either a normal diet, a high-fat diet, or a high-fat diet supplemented with SCR, ASCR, MSCR, or AMSCR. After 8 weeks, body weight gain, serum and hepatic lipid profiles, and the activities of enzymes that generate or scavenge reactive oxygen species (ROS) were evaluated. Compared with the high-fat diet group, all the test groups showed a significant reduction in body, organ, and epididymal fat weight gain. These effects were observed with supplements in the order AMSCR>ASCR>MSCR>SCR. Similarly, supplements of test samples reduced high levels of serum and hepatic triglycerides (TG), total cholesterol, and low-density lipoprotein (LDL) cholesterol caused by hight-fat diet, while high-density lipoprotein (HDL) cholesterol was increased. Interestingly, the ability of ASCR to lower serum TG was stronger than that of MSCR, while MSCR showed a stronger hypocholesterolemic effect than ASCR. Meanwhile, AMSCR returned comprehensively serum lipid levels to normal. In addition, hepatic damage was prevented with effects in the order AMSCR>ASCR>MSCR>SCR. Hepatic ROS generating system including xanthine oxidase (XO) and ROS scavenging system including superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione S-transferase (GST) were recovered to normal level by all test diets. In conclusion, this study suggests that SCR and its fermented products can inhibit obesity and improve lipid profiles.
To investigate the effect of dietary supplementation of Acanthopanax senticosus (AS) and Eucommia ulmoides (EU) on antioxidant defense system in laying hens, a total of three hundreds sixty 20-wk old Hyline brown commercial laying hens were assigned to five dietary groups for 10-wk: (1) control diet, (2) control diet supplemented with AS at 0.5%, (3) control diet supplemented with AS at 1.0%, (4) control diet supplemented with EU at 0.5% and (5) control diet supplemented with EU at 1.0%. Total antioxidant status (TAS) in blood and antioxidant enzymes including superoxide dismutase (SOD), gluthathione -S- transferase (GST) and glutathione peroxidase (GSH-Px), and lipid peroxidation in the small intestine and liver were measured. There were no changes in body weight for 10-wk dietary treatment. TAS in blood significantly (P<0.05) increased in birds fed the diet supplemented with 1% AS and 0.5 and 1.0% EU compared with those fed control diet. Especially, dietary EU showed much higher (P<0.05) TAS compared with AS. In the antioxidant defense enzymes, GST activity of the small intestine was shown to be significantly (P<0.05) increased in birds fed the diets supplemented with 0.5 and 1.0% EU compared with those fed the control diet. In addition, intestinal SOD activity significantly (P<0.05) increased in birds fed the diets supplemented with 0.5% of AS and EU. However, we could not observe any significant dietary treatment effect of those antioxidant parameters in the liver. In conclusion, dietary supplementation of 0.5% AS and EU in a laying hen diet could be applied as a potential antioxidant source to improves bio-activity of antioxidant and economical aspect in laying hens.
The preparation method of a soluble dietary fiber from oak wood (Quercus mongolica) and the effect of the soluble dietary fiber on physiological function in rat fed high cholesterol diets was investigated. The best condition for steam explosion method was 25 kgf/㎤ pressure for 6 min. The exploded samples were delignified by the filtration treatment with 1% NaOH for several times, which is the best condition. The enzymatic hydrolysis of Cellusoft cellulase was more effective than Onozuka R-10 cellulase. The manufactured soluble dietary fiber was assayed using gel permeation chromatography (GPC) and it was dissolved in water. Average molecular weight distribution of manufactured soluble dietary fiber was about 348-1,200 and it was assumed the oligomer form fraction. In order to compare the manufactured soluble dietary fiber with commercial soluble dietary fiber (pectin) on the physiological function, Sprague-Dawley male rats weighing 100$\pm$10 g were randomly assigned to one normal diet and five high cholesterol diet containing 1% cholesterol. The high cholesterol diet groups were classified to fiber free diet (FF group), 5% pectin (5P group), 10% pectin (l0P group), 5% manufactured soluble dietary fiber (5M group) and 10% manufactured soluble dietary fiber (10M group). Body weight gains in all soluble dietary fiber groups were lower than FF group. Food intakes were increased in all soluble dietary fiber groups than that of FF group. Food efficiency ratio (FER) was significantly decreased in all soluble dietary fiber groups than that of the FF group, and it was especially was highest in 10% supplemented soluble dietary fiber group. The weight of liver of the soluble dietary fiber supplemented groups were lower than those of the FF group, but weights of cecum and small intestine of all supplemented soluble dietary fiber groups were significantly increased, compared with that of FF group. The weights and water contents in feces were significantly increased by the soluble dietary fiber. The activity of the glutamic oxaloacetic transaminase in soluble dietary fiber groups were significantly decreased than those of FF group. The hepatic glutathione S-transferase activity in all soluble dietary fiber supplemented groups were higher than that of FF group. The physiological effects of the manufactured soluble dietary fiber are the same as the commercial soluble dietary fiber (pectin). The preparation method of the soluble dietary fiber from the oak chips suited to its purpose. (Korean J Nutrition 36(1) : 9~17, 2003)
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