Antioxidant enzymes are scavenger reactive-oxygen intermediates and are involved in many cellular defense systems. We previously reported that a crude extract of Garnoderma lucidum, a medicinally potent mushroom, profoundly increased the catalase gene expression and enzyme activities in mouse livers (Park et al., J. Biochem. Mol. Biol. 34. 144-149, 2001). In this study, we elucidated the detailed mechanism whereby G. lucidum stimulates the catalase activity and expression. The major active fraction was isolated from G. lucidum and methyl linoleate was considered the most major component of the fraction. In order to determine whether methyl linoleate increases mRNA and protein synthesis of catalase, Northern and Western blot analyses were performed in vivo with methyl linoleate-treated mouse liver homogenate after feeding methyl linoleate to the mice. Northern and Western blot analyses of the crude liver homogenates in the mice that were administered methyl linoleate revealed that the expression catalase was significantly increased when compared to the untreated controls. In addition, the catalase protein levels and enzymatic activities increased in the mouse liver homogenates. These results suggest that methyl linoleate that is produced by G. lucidum stimulates the catalase expression at the transcription level.
The purpose of this study is to investigate a possible contribution of nonspecific esterases, which occur in the oral cavity, to the degradation of ester bonds in polymethacrylates. One of the problems connected with the use of composite resins for restorations is their inadequate resistance to wear. It has been shown that methacrylate hydrolysis can be catalyzed by enzymes and that a carboxylic hydrolase (porcine liver esterase) catalyzed the hydrolysis of several mono - and dimethacrylates. The softening effect on a BISGMA/TEGDMA polymer induced by hydrolase will accelerate the in vivo wear of the polymer. Porcine liver esterase (EC 3.1.1.1) 3.2 mol/L $(NH_4)_2$$SO_4$ was obtained from Sigma Chemical Company. The esterase activity of one unit is defined as the amount of enzyme capable of hydrolyzing $l{\mu}mol$ ethyl butyrate per min at pH 8.0 AT $25^{\circ}C$. Phosphate buffer, 10mmol/L, pH 7.0, was made by adjustment of a solution of $Na_2HPO_4$ with $H_3PO_4$. Composite resins used in this study are Silux Plus, Z-100, Durafil VS, and Prisma APH. Cylindrical specimens, 14mm in diameter and 3mm thick, of Silux Plus, Z-100, Durafil VS, Prisma APH were polymerized under the celluloid strip. 60 specimens were divided into 2 groups. One group was emersed only in buffer solution, the other group was emersed in buffer and enzyme solution. Silux Plus and Z-100 were divided into 2 subgroups, one subgroup was cured only Visilux 2. And the other subgroup was cured Visilux 2 and Triaid II. Thereafter, specimens were polished to its best achievable surface according to manufacture's directions. The Vickers hardness of the specimens was measured after 1, 2, 4, 7, 9, 15, 50 days. The solutions were changed after each measurement. Composite resin surfaces were evaluated for the surface roughness with profilometer (${\alpha}$-step 200, Tencor instruments, USA) after 1 and 50 days. And then surfaces of specimens were pictured with stereosopy after 1 and 50 days. The results were as follows. 1. The surface hardness of Silux plus, durafil VS, and Prisma APH were decreased with time. But, the surface hardness of Z-100 was not decreased. 2. The surface hardness of all composite resins was decreased by esterase. 3. Composite resins, which were light-cured by Visilux 2 and concomitantly baked by oven, showed more hardened surface than light-cured by Visilux 2 only. 4. Significant surface changes were occured in Silux plus after esterase treatment.
This study was carried out to investigate the effect of porphyran on enzyme activity in rats and immunity in mice. Animals were divided into 5 groups, and were given porphyran diet for 4 weeks. Porphyran was extracted from Porphyra yezoensis: Diet groups were normal diet, control diet fed high fat, cholesterol and sodium cholate, control and 1% porphyran diet (1% PD), control and 5% porphyran diet (5% PD), control and 10% of porphyran diet (10% PD). Also Balb/c female mouse were injected i.p. with porphyran extract every other day for 20 days at levels of 1%, 2% and 5%. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activities were lower in the porphyran diet group than those in control group. Superoxide dismutase and catalase activities in liver homogenates were reduced in porphyran diet group compared to those of control group. Also, the level of liver thiobarbituric acid reactive substance (TBARS) was lower in porphyran group than that of control group. Porphyran increased IL-1 production in a dose-dependent manner, however, interleukine-2 production was reduced as the amount of porphyran increases. These results showed that supplementation of porphyran lowered antioxidant enzyme activities and has possibility of modulating immunological function.
Drug-metabolizing activities of Korean native cattle and swine were investigated from viewpoints of the cytochrome P-450's level, their dependent mixed function oxidase activities, the reactive oxygen species formation and cytosolic enzyme acitivities from each liver homogenates. Level of cytochrome P-450 in the liver microsome of Korean native cattle was $0.28{\pm}0.05nmole/mg$ and that in pigs $0.35{\pm}0.03nmole/mg$. Level of cytochrome $b_5$ of Korean native cattle was $0.24{\pm}0.06nmole/mg$, and that of pigs $0.2{\pm}0.05nmole/mg$, showing no difference between two species. NADPH P-450 reductase were higher in Korean native cattle ($58.3{\pm}5.3nmole/mg/min$) than in pigs ($29.9{\pm}3.8nmole/mg/min$)(p<0.01). The activities of cytochrome P-450 dependent monooxygenases such as ethoxyresorufin O-deethylase (cattle, $96.5{\pm}12.5nmole/mg/min$ ; pigs, $13.6{\pm}2.1nmole/mg/min$), N-benzphetamine N-demethylase (cattle, $5.23{\pm}0.82nmole/mg/min$ ; pigs, $0.76{\pm}0.3nmole/mg/min$) and aniline hydroxylase (cattle, $0.95{\pm}0.1nmole/mg/min$ ; pigs, $0.33{\pm}0.08nmole/mg/min$) were much higher in Korean native cattle than in swine(p<0.01). However, the activity of testosterone $7{\alpha}$-hydroxylase was higher in swine ($90.4{\pm}1.2nmole/mg/min$) than cattle (cattle, $32.8{\pm}1.2nmole/mg/min$). Interestingly, testosterone $16{\alpha}$-hydroxylase, a marker enzyme for P-450 IIA was not detected in both animal species. These results suggest that Korean native cattle and pigs have high contents of P-450 IA1 and P-450 IIIA. Total sulfhydryl compound (cattle, $10.3{\pm}1.1nmole/mg$ ; Pigs, $14.5{\pm}1.8nmole/mg$) and glutathione related enzymes except glutathione reductase (cattle, $38.1{\pm}7.9nmole/mg/min$; swine, $22{\pm}3.6nmole/mg/min$) showed higher levels in swine than in Korean native cattle. Superoxide dismutase (cattle, $7.64{\pm}0.84nmole/mg/min$ ; pigs, $4.47{\pm}0.94nmole/mg/min$) and catalase (cattle, $30.4{\pm}3.7nmole/mg/min$ ; pigs, $17.2{\pm}1.8nmole/mg/min$) were remarkably higher in Korean native cattle than in swine (p<0.05).
5,10-Methenyltetrahydrofolate synthetase from chicken liver was purified through 30-70% ammonium sulfate fractionation, Q Sepharose Fast Flow anion exchange and Source 15Phe hydrophobic interaction chromatography. Specific activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 0.0085, 0.031, 0.80 and 1.27 U/mg, respectively. Purification fold activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 1, 3.7, 94.1 and 149.4, respectively. HPLC gel permeation chromatography and SDS-polyacrylamide electrophoresis experiments indicated that the enzyme is a monomeric protein with a molecular weight of 22.8 kDa. Km for 5-methyl THF and Mg-ATP were $7.1\;{\mu}M$ and $63\;{\mu}M$, respectively. Optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. The data for metal ion specificity and stoichiometry showed that the maximum activity was obtained with a 1:l. ratio of $Mg^{2+}$. The ATP and Km values increased in the order of MgATP, MgCTP, MgUTP and MgGTP, and the maximum activities also decreased in the same order, indicating MgATP as the most efficient substrate. The enzyme was chemically modified only by tetranitrometane and 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, indicating that tyrosine and carboxylate are present in the active site.
Effects on the feeds of streptozotocin-induced diabetic rats with a giant embryonic rice on lipid peroxides level and antioxidative enzyme activites in plasma and liver tissues were investigated. Along with the experimental periods, all animals in diabetic groups had a lower increase rate in body weight than the normal control group. A giant embryonic rice-fed group showed a inhibition in the decrease of body weight, and a increase in feed intake compared to the normal control group. The organ weights of the diabetic control group were heavier than those of the normal control while rice-fed groups including the giant embryonic rice-fed group were found to have lower organ weights, and its blood sugar level was found to be lower than those of the normal group. Lipid peroxides of the giant embryonic rice-fed animals showed a lower lipid peroxidation values compared to that of the diabetic control group. Plasma vitamin A and E concentrations of the diabetic control group were significantly decreased compared to the normal control while those of the giant embryonic rice-fed group were found to be significantly higher than those of the diabetic control. Of the hepatic antioxidative enzymes, SOD activity of the giant embryonic rice-fed group was higher than that of the diabetic control group. Taken these together, low lipid peroxidation values and, in contrast, high antioxidative enzyme activities were thought to be a cause for decreasing hepatic oxidative damages.
The purpose of this study was to demonstrate the effect of Momordica Charantia L. water extracts, one of the natural chelator, on the biochemical and enzyme activity changes in the rats liver and kidney caused by lead acetate. Rat approximately 250 g in weight were grouped into the control, lead acetate treated, and the Momordica Charantia L. boiling water extracts treated after lead acetate groups. Lead acetate (1,000 ppm) and Momordica Charantia L. water extracts (5%, 10%) were delivered drinking water. Serum AST, ALT and BUN were measured, histological alteration of liver and kidney were examined by light microscopy. Momordica Charantia L. extract group was decreased serum AST, ALT and BUN level induced by lead. Optical observations of liver tissue, lead group were observed necrosis of hepatic cells and infiltration of inflammatory cells, but Momordica Charantia L. extract group was observed only slight infiltration of inflammatory cells around the central vein. Optical observations of kidney tissue, lead acetate induced atrophy and necrosis of glomerulus and infiltration of inflammatory cell around renal tubule. For the group treated with Momordica charantia L. extract, the glomerulus was similar to the control, some around the renal tubule was observed infiltration of inflammatory cells. In conclusion, Momordica Charantia L. water extract may protect the lead-induced toxicity on liver and kidney.
Hur, Sung-Ik;Park, Si-Hyang;Lee, Su-Seon;Choung, Se Young;Choi, Yeung Joon
Journal of the Korean Society of Food Science and Nutrition
/
v.42
no.12
/
pp.1940-1948
/
2013
This study is conducted to investigate the antioxidative effect of oyster hydrolysates in the serum and liver of SD-rats through the determination of lipid content, production of free radicals and antioxidant enzyme activities. Two different hydrolysates, Protamex-treated and Neutrase-treated hydrolysate with the cross-linking of protein by transglutaminase (TGPN group) and without (PN group), were fed for 6 weeks. TGPN hydrolysate in serum and liver significantly decreased the total cholesterol in the range of 26.1% to 28.9%, and triglyceride in the liver of up to 6.3%. Superoxide radical in the serum and lipid peroxide radical in the liver were significantly decreased in SD-rats fed 200 mg TGPN hydrolysate. Superoxide dismutase activity was significantly decreased in the liver of SD-rats. These results indicate that TGPN hydrolysate could scavenge the superoxide and hydroxyl radicals, and reduce the superoxide dismutase and catalase activities. The TGPN is also protected the oxidation of protein by the free radicals.
$L-Triiodothyronine(T_3)$ binding to purified plasma membrane and to isolated nuclei from the same liver in obese(ob/ob) mice and their lean littermates was examined. The maximal binding capacity(Bmax) for $T_3$ receptor of liver nuclei, as compared to lean control, was significantly lower in the obese mouse$(obese 527{\pm}80fmol/mg\;DNA ; lean 883{\pm}62fmol/mg\;DNA)$, without an apparent difference in dissociation constant(Kd). The finding that obese mice have fewer liver nuclear $T_3$ receptors confirms previous reports. The Bmax and Kd of liver plasma membrane $T_3$ receptor were not significantly different between obese and lean mouse, which suggests no defect to be occurring in the function of the plasma membrane $T_3$ receptor and reinforces the view that the peripherally impaired thyroid hormone action in obese mice is a post plasma membrane receptor event. These results further support the hypothesis that the major defect of the thyroid hormone metabolism in genetic obesity occurs at the level of the nuclear receptor.
Geun-Hui Nam;Young-Bin Lee;Sea-Hwan Sohn;In-Surk Jang
Korean Journal of Poultry Science
/
v.51
no.2
/
pp.107-116
/
2024
The study was conducted to examine age-related development of digestive organs, intestinal microvilli hydrolase, and hepatic antioxidant enzyme in Korean native chicks (KNC) aged from 0-d to 28-d of post-hatching. Body weight did not significantly increase from 0-d to 3-d-old, but after that remarkably increased from 3-d to 28-d-old (P<0.05). The relative weight (g/100 g of BW) of the proventriculus, gizzard, and liver was significantly higher at 3- and 7-d-old chicks than that of the other ages. The relative weight of the intestine, mucosal tissues, and pancreas was markedly developed at the ages of 3-, 14-, and (or) 21-d-old chicks (P<0.05). In the small intestine, the specific activities of maltase and sucrose were significantly higher at 14-d-old compared with the other ages (P<0.05). Leucine aminopeptidase activity showed a constant level from 0- to 28-d-old without significance. The specific activity of alkaline phosphatase was significantly higher at 0-d-old compared with the other ages (P<0.05). In the liver, the specific activities of superoxide dismutase, glutathione peroxidase, and glutathione S-transferase were shown to be lowest at 0-d-old, but they continued to increase as the age increased. The lipid peroxidation was significantly high at the age of 21-d (P<0.05), after that its level decreased at 28-d old. In conclusion, the KNC rapidly developed digestive organs and intestinal microvilli hydrolase activity from 3- to 14-d-old after hatching. Hepatic antioxidant enzyme activity continued to increase as the age increased after hatching, resulting in 28-d-old chicks showing the highest antioxidant enzyme activity in the KNC.
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