• Journal of Veterinary Clinics
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• v.29 no.5
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• pp.372-376
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• 2012

Acute kidney injury (AKI) is a serious problem associated with high morbidity and mortality. Ischemia-reperfusion is an important cause of acute kidney injury. This study was performed to ascertain clinically useful biomarkers for the diagnosis of AKI. In three miniature pigs, AKI were induced by 60 minutes of bilateral renal ischemia by the clamping renal artery. Blood and urine samples were collected from the pigs prior to clamping (baseline) and 0, 1, 3 and 5 days post-clamping. Serum blood urea nitrogen (BUN), creatinine, sodium and uric acid were measured in serum and urine samples. Fractional excretion of sodium ($FE_{Na}$) and fractional excretion of uric acid ($FE_{UA}$) were calculated. Also, interleukin (IL)-6, IL-18, liver type fatty acid binding protein (L-FABP) and glutathione-S-transferase (GST) were detected by Western immunoblotting. Serum BUN and creatinine levels were increased significantly at day 1 post-clamping in all three miniature pigs. However, $FE_{Na}$ and $FE_{UA}$ showed marked individual differences. Western immunoblotting revealed significantly increased levels of IL-6, IL-18, L-FABP and GST in post-ischemic urine, compared to pre-clamping. While more research concerning the variance of $FE_{Na}$ and $FE_{UA}$ is needed, serum BUN, creatinine, IL-6, IL-18, L-FABP and GST may be sensitive urine biomarkers for diagnosis of AKI together with other biomarkers in the porcine ischemia-reperfusion model.

• 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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.1
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• pp.81-86
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• 2002

The present study was conducted to investigate the effect of dietary supplementation of vitamin A or $\beta$-carotene on oxidative damage induced by acute ethanol administration. Sprague-Dawley rats were fed on the experimental diets supplemented with retinyl acetate (2.86 mg/kg diet) or $\beta$-carotene (15.2 mg/kg diet) for 5 weeks. After fed the diet, rats were administered 20% ethanol solution (3g/kg B.W.) acutely. Lipid peroxide values in hepatic tissue, hepatic antioxidative enzyme activities and contents of antioxidative nutrient such as vitamins A and E in serum and hepatic tissue were measured. Hepatic level of malondialdehyde decreased in $\beta$-carotene group compared to the control group. However, there was no significant difference between retinal acetate and $\beta$-carotene groups. Superoxide dismutase activity was higher in retinal acetate group than in the control group. Hepatic glutathione-S-transferase activity of retinal acetate and $\beta$-carotene groups significantly decreased as compared with that of control group. The hepatic content of retinol increased in retinal acetate and $\beta$-carotene groups, especially, in retinyl acetate group. But there was no significant difference in serum content of retinol among the groups. Hepatic content of $\alpha$-tocopherol was significantly increased in retinyl acetate and $\beta$-carotene groups. In conclusion, acute ethanol administration might induce lipid peroxidation, and the dietary supplementation of retinyl acetate or $\beta$-carotene improve partly the antioxidative system through activation of superoxide dismutase and retention of hepatic $\alpha$-tocopherol in ethanol-treated rats.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.8
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• pp.1010-1015
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• 2006

p-Pheylenediamimine (PPD) is one of hair dye's ingredients, and the mixture of PPD with hydrogen peroxide is generally used to dye hair at beauty shop. This study is conducted to investigate the effect of oxidized PPD on rat skin. 6% hydrogen peroxide, PPD (5% PPD in 2% $NH_4OH$) or the mixture (isovolumed mixture of 5% PPD and 6% hydrogen peroxide in 2% $NH_4OH$) was applied to rat skin ($25\;mg/16.5\;cm^2$) five times every other day. The activity of acid phosphatase (ACP) was more increased in the mixture of PPD with hydrogen peroxide applied group than PPD applied group. Furthermore, the activity of glucose 6-phosphatase (G6Pase) in the mixture of PPD with hydrogen peroxide applied group showed higher decreasing rate than that of PPD applied group. In histopathological findings, the mixed PPD with hydrogen peroxide applied group showed more thickening of epithelium, increased numbers of dermal fibroblasts, and the dilatation of dermal capillaries than PPD applied group. The significant increasing of xanthine oxidase (XO) activity was determined in mixture of PPD with hydrogen peroxide applied group compared with PPD applied group. However, reactive oxygen species (ROS) scavenging system, the activities of superoxide dismutase (SOD) and glutathione S-transferase (GST) were more significantly decreased in mixed PPD with hydrogen peroxide applied groups than in PPD applied group. In conclusion, topical application with the mixture of PPD with hydrogen peroxide compared with PPD application resulted in imbalance with ROS generating and scavenging which probably led to severe skin injury.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• Journal of Life Science
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• v.20 no.7
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• pp.1005-1011
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• 2010

$\gamma$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system. GABA transporters (GATs) control extracellular GABA levels by reuptake of released GABA from the synaptic cleft. However, how GATs are regulated has not yet been elucidated. Here, we used the yeast two-hybrid system to identify the specific binding protein(s) that interacts with the carboxyl (C)-terminal region of GAT1, the major isoform in the brain and find a specific interaction with the ubiquitin-specific protease 14 (Usp14), a deubiquitinating enzyme. Usp14 protein bound to the tail region of GAT1 and GAT2 but not to other GAT members in the yeast two-hybrid assay. The C-terminal region of Usp14 is essential for interaction with GAT1. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to GAT1 specifically co-immunoprecipitated Usp14 from mouse brain extracts. These results suggest that Usp14 may regulate the number of GAT1 at the cell surface.

• Korean Journal of Medicinal Crop Science
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• v.15 no.6
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• pp.437-443
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• 2007

This study was carried out to investigate the effect of dietary supplementation with red ginseng water-extracts on the induction of microsomal cytochrome P-450 in rats. Phenobarbital (PB) and 3-methylcholanthrene (3-MC), P-450 inducers, were administered to 3- or 12-month old rats received red ginseng extracts (25 mg/kg) from 6 weeks to 12 months for 3 days. PB and 3-MC increased levels of P-450, P-450 reductase, ethoxycoumarin O-deethylase, benzphetamine N-demethylase and glutathione-S-transferase in the liver of rats. However, chronic administration of red ginseng significantly reduced these increase of enzyme levels induced by P-450 inducers. Chronic administration of red ginseng did not affect the induction of cytochrome $b_5$ and NADH cytochrome $b_5$ reductase by P-450 inducers. It is suggested that the induction of cytochrome P-450 system in the liver in relation to xenobiotics toxicity can be modulated by long-term supplementation with Korean red ginseng to rats.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.11
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• pp.1286-1292
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• 2017

This study was conducted to investigate the hepatoprotective effects of 3,5-dicaffeoylquinic acid (3,5-DCQA) isolated from Ligularia fischeri against hydrogen peroxide-induced oxidative stress in HepG2 cells. Antioxidative effects of 3,5-DCQA were determined by measuring antioxidant enzyme [superoxide dismutase (SOD), catalase (CAT) glutathione peroxidase (GPx)] expression levels against hydrogen peroxide-induced oxidative stress using real-time PCR analysis. 3,5-DCQA treatment significantly increased gene expression levels of SOD, CAT, and GPx in a dose-dependent manner ($10{\sim}30{\mu}g/mL$) in HepG2 cells. Hepatoprotective effects were analyzed by measuring glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) activities using a biochemistry analyzer in hydrogen peroxide-treated HepG2 cells. 3,5-DCQA treatment significantly reduced GOT, LDH, and GGT activities in a dose-dependent manner ($10{\sim}30{\mu}g/mL$) against increased liver function index enzyme activities induced by hydrogen peroxide oxidative stress in HepG2 cells. The results reveal that 3,5-DCQA compound isolated from Ligularia fischeri can be useful for the development of an effective hepatoprotective agent.

• Korean Journal of Food Science and Technology
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• v.34 no.4
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• pp.684-687
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• 2002

Water and methanol extracts of cricket were examined for their liver protective effects against $CCl_4-intoxication$ in ICR-mice. Serum transaminases (S-GOT and S-GPT), lactate dehydrogense (LDH) and glutathione-S-transferase (GST) activities and TBARS (Thiobarbiturate-reactive substances) content were measured for evaluation of liver protective effects. The activities of GOT, GPT, LDH and hepatic content of lipid peroxide after $CCl_4-treatment$ were higher than normal control but those levels decreased th 74, 50, 101 and 40%, respectively, by the treatment of cricket methanol extract. The anti-fatigue effects of water and methanol extracts investigated by an acute weight-loaded forced swimming test showed significantly prolonged swimming time in the mice administered cricket extracts. These results suggest us that water/alcohol extract of G. bimaculatus may be used as a liver protective food.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.928-932
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• 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.