This study was done to investigate effects of ${\gamma}$-irradiated pork feeding on the formation of glutathione S-transferase placental form positive (GST-P$^{+}$) foci, lipid peroxidation, cytochrome P450 system and microsomal glucose 6-phosphatase activity in diethylnitrosamine (DEN)-initiated rat hepatocarcinogenesis. Weaning Sprague-Dawley male rats were fed the diet containing ${\gamma}$-irradiated ground pork at the dose of 0, 3, 10, 30 kGy as a 20% of protein source for 8 weeks. One week after feeding, rats were intraperitoneally injected twice with a dose of DEN (50 mg/kg BW). As a promote.,0.05%phenobarbital was fed in drinking water from one week after DEN treatment until the end of experiment. At the end of 8th week, rats were sacrificed and hepatic GST-P$^{+}$ foci, microsomal malondialdehyde (MDA) and conjugated diene contents were determined. In addition, cytochrome P450 content and the activities of NADPH cytochrome P450 reductase and glucose 6-phosphatase were also measured. There was no significant effect by gamma irradiation on microsomal MDA content, conjugated diene, cytochrome P450 content and activities of NADPH cytochrome P450 reductase and glucose 6-phosphatase. However with DEN treatment, microsomal MDA content showed a increasing tendency. Cytochrome P450 content was also significantly increased while microsomal glucose 6-phophatase activity was significantly decreased with DEN treatment. However the activity of NADPH cytochrome P450 reductase was not affected. An interesting finding in this study was that the number and area of hepatic GST-P$^{+}$ foci of rats fed gamma irradiated pork were tended to be decreased by high dose of irradiation, but were not significantly different. These results might imply that the consumption of low dose of gamma irradiated pork does not affect the formation of hepatic GST-P$^{+}$ foci and lipid peroxide and membrane stability.ability.
This study was done to investigate effects of ${\gamma}$ irradiated beef feeding on the formation of gluta thione S transferase placental form positive(GST P+) foci, lipid peroxidation, cytochrome P450 system and microsomal glucose 6 phosphate activity in diethylnitrosamine(DEN) initiated rat hepatocarci nogenesis. Weaning Sprague Dawley male rats were fed the diet containing ${\gamma}$ irradiatied ground beef at the dose of 0, 3, 5kGy as a 20% of protein source for 8 weeks. One week after feeding, rats were intraperitoneally injected twice with a dose of DEN(50mg/kg BW). As a promoter, 0.05% phenobarbital was fed in drinking water from one week after DEN treatment until the end of experiment. At the end of 8th week, rats were sacrificed and hepatic GST P+ foci, microsomal malondialdehyde(MDA) and conjugated diene contents were determined. In addition, cytochrome P450 content and the activities of NADPH cytochrome P450 reductase and glucose 6 phosphatase were also measured. There was no significant effect by gamma irradiation on microsomal MDA content, conjugated diene, cytochrome P450 content and activities of NADPH cytochrome P450 reductase and glucose 6 phosphatase. However with DEN treatment, microsomal MDA content and conjugated diene contents were significantly changed. Cytochrome P450 content was also significantly increased while microsomal glucose 6 phophatase activity was significantly decreased with DEN treatment. However activity of NADPH cytochrome P450 reductase was not affected. An interesting finding in this study was that the number and area of hepatic GST P+ foci of the rats fed gamma irradiated beef were significantly(p<0.05) lower than those of the control. Such a lowering effect on GST P+ foci formation was highest at the dose of 3kGy than others. Overall results suggest that the consumption of low dose of gamma irradiated beef does not affect the formation of lipid peroxide, cytochrome P450 system and membrane stability.
Adriamycin is a commonly used chemotherapeutic agent for cancer, including acute leukemia, lymphoma, and a number of solid human tumors. However, recent studies have recognized severe cardiotoxicity after an acute dose, which are likely the result of generation of free radicals and lipid peroxidation. Therefore, the clinical uses of adriamycin have been limited. Melatonin, the pineal gland hormone known for its ability to modulate circardian rhythm, has recently been studied in its several functions, including cancer growth inhibition, stimulating the immune system, and acting as an antioxidant and radical scavenging effects. In the present study, we evaluated the effect of melatonin administration on adriamycin-induced cardiotoxicity in rat. Heart slices were prepared using a Stadie-Riggs microtome for the measurement of malondialdehyde (MDA) content used as an index of lipid peroxidation and lactate dehydrogenase (LDH) release as an indicator of lethal cell injury. Serious adriamycin-induced lethality was observed in rat by a single intraperitoneal injection in a dose-dependent manner. A single injection of adriamycin (25 mg/kg, i.p.) induced a lethality rate of 86%, with melatonin (10 mg/kg s.c. for 6 days) treatment reducing the adriamycin-induced lethality rate to 20%. The severe body weight loss caused by adriamycin was also significantly attenuated by melatonin treatment. Treatment of melatonin marked reduced adriamycin-induced the levels of MDA formation and LDH release. A cell damage indicated by the loss of myofibrils, swelling of the mitochondria as well as cytoplasmic vacuolization was seen in adriamycin-treated group. Melatonin attenuated the adriamycin-induced structural alterations. These data provide evidence that melatonin prevents adriamycin-induced cardiotoxicity and might serve as a combination with adriamycin to limit free radical-mediated cardiotoxicity.
The antioxidative effect, antimutagenic capacity and inhibitory effect of cancer cell proliferation in ethanol extracts of Ganoderma lucidum were studied for suggestion of prevention and dietetic treatment of chronic diseases and development of antioxidative, antimutagenic and anticancer functional food by employing biological and biochemical assay. The IC/sub 50/ of MDA with BSA conjugation reaction, lipid peroxidation and scavenging effect on DPPH radical in ethanol extracts of Ganoderma lucidum showed 1026.21 mg/mL, 0.152 mg/mL and 0.412 mg/mL, respectively. So, the most effective antioxidative capacity in ethanol extracts of Ganoderma lucidum was the lipid peroxidation, among the method used this study. The indirect and direct antimutagenic effects of ethanol extracts of Ganoderma lucidum were examined by Ames test using Salmonella typimurium TA98 and TA100. The inhibition rates on indirect mutagenicity mediated by 2-Anthramine showed 100% in the Salmonella typimurium TA98 and 98.74% in the Salmonella typimurium TA100 and the direct mutagenicity mediated by sodium azide in Salmonella typimurium TA100 was 82.96%. But, the inhibitory effect on indirect mutagenicity mediated by 2-Nitrofluorene in Salmonella typimurium TA98 was low(7.81%). The inhibitory Effect of Ganoderma lucidum ethanol extracts on cell proliferation in Hela and MCF-7 by MTT test were 74.36% in HeLa and 73.90% in MCF-7 at the 0.50 mg/assay concentration and IC/sub 50/ were 0.163 mg/mL and 0.196 mg/mL respectively. From this result, it is suggested that Ganoderma lucidum is believed to have a possible antioxidative, antimutagenic and anticancer capacities.
This study was conducted to compare the effects of n-6 linoleic acid and n-3 linolenic acid on lipid peroxidation and the activities of enzymes defending against oxidation, which are involved in the tumor promotion, and histolOgical changes of hepatocarcinogen treated rat liver. In this study, weanling male Sprague-Dawley rats were fed one of three diets, containing 15% (w/w) of beef fat (BF), com oil (CO) or perilla oil (PO), for 11 weeks. During the 3rd week, experimental groups were injected with 2-AAF (50 mg/kg of BW) intraperitoneally 3 times. Findings show that the com oil diet group has greater liver MDA content than the beef fat and perilla oil diet groups. Also, it is observed that the perilla oil diet group has lower MDA content than beef fat and com oil diet groups, even though perilla oil is more desaturated than beef fat and com oil. In terms of activity, mixed-function oxidase activity is not Significantly affected by the different dietary fats and 2-AAF treatment. GSH-peroxidase, GSH-reductase and GSH-Stransferase activities are significantly higher in the CO+AAF group than those of the other groups. GST and GSH-Px are activated by 2-AAF treatment in the com oil diet group only. The hepatocytes of the BF+AAF group were the most severely degenerated, the second was the CO+AAF group and the least was the PO+AAF group. It was also found that dietary com oil increased lipid peroxidation and activated defense enzymes against oxidation in liver, but dietary perilla oil did not, or supressed defense enzymes. Therefore it is concluded that dietary n-3 linolenic acid in perilla oil inhibits lipid peroxidation and carcinoenesis in rat liver following 2-AAF treatment.
Objective : This study was undertaken to examine whether Carthami Semen aquacupuncture (CSA) exerts protective effect against Hg-induced cell injury in rabbit liver. Methods : The cell injury was evaluated by ALT activity and lipid peroxidation was estimated by measuring malondialdehyde (MDA). Results : Hg caused an increase of ALT activity and lipid peroxidation in a dose-dependent-manner over concentrations of 0.1-1 mM, which were prevented by addition of 0.005% CSA. The protective effect of CSA was dose-dependent in concentration range of 0.001 to 0.01%. The increase of ALT activity and lipid peroxidation induced by 0.5 mM Hg were almost completely decreased by addition of 0.01% CSA. When the liver tissues were exposed to 0.5 mM Hg, GSH content was decreased, which was significantly restored by 0.01% CSA. 0.5 mM Hg caused decrease in the amount of total and nonprotein sulfhydryl groups, and 0.01% CSA prevented Hg-induced reduction of nonprotein sulfhydryl group but not protein sulfhydryl group. Conclusions : These results suggest that CSA exerts protective effect against Hg-induced cell injury by antioxidant action resulting from enhancement of nonprotein sulfhydryl group content including GSH in liver.
Objectives : The purpose of this study is to examine the antioxidant effects on male mouse reproductive cells of the extract of Platycladi Semen. Methods : The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, cell viability by a modified MTT assay, the effects on $H_2O_2$-induced cytotoxicity by MTT assay, lipid peroxidation by malondialdehyde (MDA) formation and super oxide dismutase (SOD), respectively. Results : The results showed that the extract scavenged DPPH radical in a dose-dependent manner by up to 74.87%. The cell viability of the extract was within 72~96% on Leydig cells and GC-2 cells at concentrations of 1, 5, 10, 50 and 100 ug/ml. The hydrogen peroxide-induced cytotoxicity of Leydig cells was protected to 72.09% by the extract at concentration of 100 ${\mu}g/ml$. The hydrogen peroxide-induced lipid peroxidation of MDA formation was decreased to 1.80 and 1.65 nmoles/mg protein by the extract at concentrations of 50 and 100 ${\mu}g/ml$. The extract at all concentrations, SOD activity was not significantly changed. Conclusions : In conclusion, the extract of Platycladi Semen has antioxidant effects on Leydig cells and protect male reproductive system against oxidative stress.
The purpose of this study was to determine the extent of oxidative stress in NIDDM patients with diabetic complications and to determine the relationship between oxidative stress and diabetic complications. For this study, 139 NIDDM patients were recruited, 85 with diabetic complications and 54 without complications were recruited. The concentration of malondialdehyde(MDA) and the activities of antioxidant enzymes including catalase, superoxide dismutase(SOD), gluthatione peroxidase(GSH-Px)were determined. The daily intakes and plasma concentrations of beta-carotene, lycopene, lutein nd alpha-tocopherol were determined by food frequency questionnaire and by high performance liquid chromatography(HPLC), respectively. Among the antioxidant enzymes studied, only GSH-Px activity was lower in NIDDM patient, with diabetic complications than in those without complications(2.91$\pm$0.80 vs 3.54$\pm$0.44 U/mgHb, p<0.05). Those NIDDM patients with diabetic complications had higher MDA concentrations than those without diabetic complications(1.40$\pm$0.25 vs 1.25$\pm$0.11 nmol/ml, p<0.05). There were no significant differences in the dietary intakes of total carotenoids(2854 vs 2824ug/day)or vitamin E (9.5$\pm$3.2 vs 9.5$\pm$2.0mg/day)between NIDDA patients with and without complications. However, the plasma concentrations of beta-carotene and lycopene were significantly lower in NIDDM patients with complications than in NIDDM patients without complications (Beta-carotene : 24.2$\pm$12.5 vs 33.1$\pm$16.2(ug/dl), lycopene : 2.8$\pm$2.1 vs 4.3$\pm$2.8(ug/dl)). This study showed that in NIDDM patients with complications, the lipid peroxidation of erythrocytes was higher increased and the antioxidant reserves were significantly dipleted, compared with NIDDM patients without complications. The lower plasma concentrations of beta-carotene and lycopene in NIDDM patients may be due to the presence of diabetic complication, not due to the lower dietary intakes of antioxidant vitamins. To define the role of carotenoids in diabetes, more experimental and clinical studies are needed.
Malondialdehyde (MDA) and total antioxidant capacity (T-AOC) levels, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), glutathione transferase (GST) and xanthine oxidase (XOD) activities were analyzed in serum, livers and kidneys of pigs treated with graded doses of fluoride (as NaF). Ninety-six Duroc-Landrace-Yorkshire crossbred growing pigs (48 barrows and 48 gilts, respectively), with similar initial weight 24.14${\pm}$1.12kg, were randomly assigned to four different treatments. These treatments containing the following added F: basal control; 50 mg/kg F; 100 mg/kg F and 150 mg/kg F were randomly assigned to four pens (three barrows and three gilts) each in a completely randomized design. The results showed pigs treated with 150 mg/kg F significantly decreased average daily gain (ADG) (p<0.05) and increased feed/gain ratio (F/G) (p<0.05) compared to the controls. In the groups treated with fluoride, the contents of MDA increased, T-AOC levels and the activities of SOD, GSH-PX, CAT, GST and XOD decreased, and most of which altered significantly (p<0.05). The study therefore indicated the mechanism of excess fluoride on the impairment of soft tissues involved in lipid peroxidation and decreased the activities of some enzymes associated with free radical metabolism.
This study investigated the protective effect of the butanol (BuOH) fraction from fermented Laminaria japonica extract (BFLJ) on AAPH-induced oxidative stress in porcine kidney epithelial cells (LLC-PK1 cells). L. japonica was fermented by Aspergillus oryzae at $35{\pm}1^{\circ}C$ for 72 h. Freeze-dried fermented L. japonica was extracted with distilled water, and the extracted solution was mixed with ethanol and then centrifuged. The supernatant was subjected to sequential fractionation with various solvents. The BuOH fraction was used in this study because it possessed the strongest antioxidant activity among the various solvent fractions. The BuOH fraction of fermented L. japonica had a protective effect against the AAPH-induced LLC-PK1 cells damage and increased cell viability while reducing lipid peroxidation formation and increased activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. The inhibitory effect of BFLJ on lipid peroxidation formation had a higher value of $0.11{\pm}0.01nmol$ MDA at $100{\mu}g/mL$ concentration in comparison with intact BuOH fraction showing $0.22{\pm}0.08nmol$ MDA at the same concentration. Furthermore, BFLJ treatment increased glutathione concentration. GSH concentration in the cell treated with BFLJ of $100{\mu}g/mL$ was $1.80pmol/L{\times}10^5cells$. These results indicate that BFLJ protects the LLC-PK1 cells against AAPH-induced cell damage by inhibiting lipid peroxidation formation and increasing antioxidant enzyme activities and glutathione concentration.
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