mRNA Expression Levels Of Superoxide Dismutase Research Articles

Effects of Dietary Eucommia ulmoides Leaf Extract on Growth, Muscle Composition, Hepatopancreas Histology, Immune Responses and Microcystin-LR Resistance of Juvenile Red Claw Crayfish (Cherax quadricarinatus)

The objective of the present study was to investigate the effects of different doses of dietary Eucommia ulmoides leaf extract (ELE) on juvenile red claw crayfish (Cherax quadricarinatus). A total number of 720 red claw crayfish (initial body weight of 0.24 ± 0.01 g) were randomly assigned to six groups and fed diets containing 0 (Diet 1), 0.5 (Diet 2), 1 (Diet 3), 2 (Diet 4), 4 (Diet 5) and 10 (Diet 6) g dry weight (dw) ELE kg (dw)−1 diets for eight weeks and challenged with microcystin-LR stress. The results indicated that dietary supplementation with 1–2 g dw ELE kg (dw)−1 diet could significantly improve the weight gain rate (WGR) and specific growth rate (SGR) of crayfish. Muscle crude protein contents of crayfish fed Diet 2, Diet 3, and Diet 4 were significantly higher than those of the control group. Compared with the control group, dietary ELE could increase total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GPx), acid phosphatase (ACP), alkaline phosphatase (AKP), and phenoloxidase (PO) activities and decrease malondialdehyde (MDA) level of crayfish. Dietary ELE significantly increased the relative expression levels of SOD, thioredoxin 1 (TRX1), GPx, selenium-dependent glutathione peroxidase (Se-GPx), cytochrome P450 (CYP450), anti-lipopolysaccharide factor (ALF) and C-type lysozyme (C-LZM) mRNA of crayfish compared with the control group during the feeding experiment. When subjected to MC-LR stress for 48 h, the mRNA expression levels of SOD, GPx, Se-GPx, glutathione-s-transferase 1 (GST1), ALF, hemocyanin (HEM), and C-LZM in the hepatopancreas could be improved to varying degrees compared with the Diet 1. Supplementation of 1–2 g dw ELE kg (dw)−1 diet could improve the survival rate (SR) of crayfish under MC-LR stress. These results indicated that dietary ELE (1–2 g dw ELE kg (dw)−1 diet) could improve the growth performance, muscle protein, and non-specific immune response and increase the SR of crayfish under MC-LR stress by regulating the mRNA expression levels of the immune- and antioxidant-related genes.

Protective effect of Insect tea primary leaf (Malus sieboldii (Regal) Rehd.) extract on H2O2-induced oxidative damage in human embryonic kidney 293T cells

In this study, Insect tea primary leaf (Malus sieboldii (Regal) Rehd.) was used as the research object to investigate the protective effect of Insect tea primary extract (ITPLE) on hydrogen peroxide (H2O2)-induced oxidative damage in human embryonic kidney 293T cells (HEK 293T cells) and the mechanism of action of the main active components. The 3-(4,5-dimethyl-2-thiazolyl)- 2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to determine the toxicity of ITPLE to HEK 293T cells in vitro as well as its protective effect against (H2O2)-induced oxidative damage in HEK 293T cells. In addition, various assay kits were used to measure oxidation-related indicators in HEK 293T cells, and quantitative polymerase chain reaction (qPCR) analysis was used to determine the mRNA expression levels of oxidation-related genes in HEK 293T cells. High performance liquid chromatography (HPLC) analysis was used to characterize active components in ITPLE. The experimental results revealed that the ITPLE had no toxic effect on cells in the range of 0–200 μg/mL, and, in this range, exhibited a concentration-dependent protective effect against H2O2-induced oxidative damage in HEK 293T cells. It was also found that the ITPLE can reduce the malondialdehyde (MDA) level and increase the levels of superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px), and catalase (CAT)in oxidative damage HEK 293T cells. The qPCR analysis results also showed that the ITPLE upregulated the mRNA expression levels of SOD, CAT, GSH and GSH-Px in HEK 293T cells damaged by H2O2-induced oxidative stress. The HPLC analysis identified 7 bioactive components in the ITPLE, including neochlorogenic acid, cryptochlorogenic acid, rutin, kaempferin, isochlorogenic acid B, isochlorogenic acid A and hesperidin. This study reveals that ITPLE is rich in active compounds and has good antioxidant effect in vitro, thus it has the potential to be developed into a traditional Chinese medicine and functional drinks.

Effects of dietary rumen-protected betaine supplementation on the antioxidant status of lambs

As a methyl group donor and organic osmolyte, betaine has been widely used in livestock production, but the antioxidant defence properties of betaine in lambs have not been determined. The objective of this study was to investigate the effect of dietary betaine and rumen-protected betaine (RPB) supplementation on the growth performance and antioxidant status of lambs. A total of sixty male Hu sheep (approximately 30 kg body weight) were randomly assigned into five treatments and fed basal diets (Con group), basal diets with 2.2 g/d betaine supplementation (B-2.2 group), basal diets with 2 g/d RPB supplementation (RPB-2 group), basal diets with 4 g/d RPB supplementation (RPB-4 group), and basal diets with 6 g/d RPB supplementation (RPB-6 group) for 70 d. After the feeding experiment, the antioxidant indexes in the serum, Longissimus dorsi (LD) and liver were measured. Dietary supplementation of RPB increased the average daily gain (ADG) and decreased the feed-to-gain ratio (F/G) of lambs (P < 0.05), whereas treatments had no significant effect on the average daily feed intake (ADFI). The activity of glutathione peroxidase (GSH-Px) in the serum was increased in the RPB-2 group (P < 0.05); the activities of superoxide dismutase (SOD) and catalase (CAT) in the LD were increased in the RPB-4 group (P < 0.05);and the total antioxidant capacity (T-AOC) and the activities of SOD, CAT and GSH-Px in the liver were all elevated in the B-2.2 group (P < 0.05). The concentrations of malondialdehyde (MDA) in the serum, LD and liver decreased in the RPB-2 group (P < 0.05). In addition, the mRNA expression levels of SOD, CAT, nuclear-related factor 2 (Nrf2), Kelch-like ECH-associated protein1 (Keap1) and peroxiredoxin 3 (PRDX3) in the LD were increased in the B-2.2 group (P < 0.05), and the mRNA expression levels of CAT, Keap1 and PRDX3 in the liver were increased in the RPB-2 group (P < 0.05).Considering both growth performance and antioxidant status, 2.0 g/d RPB is recommended as a supplement for lambs.

Glutamine improves heat stress–induced oxidative damage in the broiler thigh muscle by activating the nuclear factor erythroid 2–related 2/Kelch-like ECH-associated protein 1 signaling pathway

The aim of the present study was to evaluate the effect of glutamine (Gln) on modulating heat stress–induced oxidative damage in the broiler thigh muscle through nuclear factor erythroid 2–related 2/Kelch-like ECH-associated protein 1 (Nrf2-Keap1) pathway. Three-hundred 22-day-old Arbor Acres broilers were reallocated into 5 groups: a control group (24 °C) fed with basal diet and 4 heat stress (HS) groups (34 °C for 8 h/D) fed with basal diet containing 0, 0.5, 1.0, and 1.5% Gln. This experiment lasted 21 D. Heat stress decreased (P < 0.05) pH, redness, and Gln levels, and increased (P < 0.05) luminance, water loss rate, and cooking loss (CL) values of the thigh meat. Compared with the HS group, supplementation with 1.5% Gln increased (P < 0.05) pH, redness, and Gln levels, but decreased (P < 0.05) luminance and CL values in the thigh meat. There were significant decreases (P < 0.05) in glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), and Nrf2 levels, but significant increases (P < 0.05) in the malondialdehyde (MDA) and Keap1 levels of the thigh muscle after HS treatment. Compared with the HS group, supplementation with 1.0, and 1.5% Gln decreased (P < 0.05) MDA and Keap1 levels; supplementation with 1.5% Gln increased (P < 0.05) GSH, GSH-Px, T-AOC, CAT, SOD, and Nrf2 levels in the thigh muscle of heat-stressed broilers. Furthermore, HS decreased (P < 0.05) Nrf2, SOD, CAT, and GSH-Px mRNA expression levels, but increased (P < 0.05) Keap1 mRNA level in the thigh muscle of broiler. Dietary supplementation with 1.5% Gln increased (P < 0.05) Nrf2, GSH-Px, CAT, and SOD mRNA expression levels, but decreased (P < 0.05) Keap1 mRNA level in the thigh muscle of heat-stressed broilers. In conclusion, dietary Gln improved the resistance of heat-stressed broiler muscles to oxidative damage possibly through reversing the muscle Gln level and inducing the expression of the Nrf2-Keap1 pathway.

Ecotoxicological effects of transgenic mCry1Ac maize (BT799) on zebrafish

The safety of feed derived from genetically modified (GM) crops is one of the focuses of attention. To evaluate the ecotoxicological effects of transgenic mCry1Ac maize (BT799) on fish, zebrafish (Danio rerio) were fed extruded feeds containing either 20% GM maize (GMF) or its parental control maize (PF), GM maize meal (GMM) or its parental control maize meal (PMM), and a control commercial feed (CF), respectively. The growth performance, histopathology, reproduction, antioxidant enzyme activity and mRNA expression levels of sensitive protein in the liver were investigated over the course of a 98-day feeding trial. The results showed that transgenic mCry1Ac maize had no significant effect on growth, histopathology of the liver, brain and intestinal tract, fecundity, hatching rate of fertilized eggs, superoxide dismutase (SOD), catalase (CAT) activity, mRNA expression levels of SOD and CAT, or heat shock protein 70 (HSP70) and vitellogenin (VTG) in the liver. However, zebrafish fed the commercial feed exhibited significantly greater weight, longer length, and higher specific growth rate than those fed feeds (GMF and PF) and maize meals (GMM and PMM). The hatching rate of zebrafish in the feed groups was significantly lower than that of the maize meal groups and the commercial feed group. The mRNA transcriptional levels of VTG were significantly higher in the liver for the feed groups (3.85±0.76) than that for the maize meal groups (1.60±0.56). These results suggest that transgenic mCry1Ac maize has no ecotoxicological effects on zebrafish. However, the differences in nutrient composition and palatability between the extruded experimental feeds and the commercial feed would lead to significant diffe-rences in some parameters.

Tissue-specific changes in OGG1 and SOD mRNA expression caused by NaOCl exposure in black seabream (Acanthopagrus schlegelii)

The DNA-damage defense mechanism was studied in black seabreams after oxidative stress caused by exposure to sodium hypochlorite (NaOCl). Liver, muscle, and brain tissues were obtained after different NaOCl-exposure times (0, 24, 48, 72, and 96 h) and concentrations (0.5, 1, 1.5, 2, and 3 mg/L), after which oxoguanine glycosylase (OGG1) and superoxide dismutase (SOD) mRNA-expression levels were analyzed. At all NaOCl concentrations tested, liver OGG1 expression increased to a maximum in a time-dependent manner after NaOCl exposure and then decreased. In muscles, OGG1 expression increased over time following exposure to a low concentration of NaOCl (0.5, 1, and 1.5 mg/L), whereas it showed a mixed pattern (both increases and decreases observed) in the high-concentration groups (2 and 3 mg/L). SOD mRNA expression increased over time, both in the liver and muscles. In the brain, both OGG1 and SOD mRNA expression levels were highest after exposure to the lowest NaOCl concentration (0.5 mg/L), whereas basal levels were maintained over time at higher concentrations. These results indicate that OGG1 and SOD provide resistance to oxidative stress in black seabreams. In addition, continuous exposure to oxidative stress can suppress enzyme expression, suggesting a risk for long-term exposure to NaOCl.

Wheat bran feruloyl oligosaccharides modulate the phase II detoxifying/antioxidant enzymes via Nrf2 signaling

The antioxidant activities of wheat bran feruloyl oligosaccharides (FOs) were determined in rats by determining the activities and mRNA expression levels of phase II detoxifying/antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and heme oxygenase-1 (HO-1) in rat organs. FOs was given by gavage at doses of 0.25, 0.5, and 0.75mmol/kg body weight every day for 15 days. Compared with the control group, the activities of SOD, CAT, and GSH-Px in FOs treatment groups significantly (P<0.05) increased in heart, liver, and kidney. All the FOs treatment also significantly (P<0.05) increased the glutathione (GSH) contents in heart (28–58%), liver (32–71%), and kidney (31–73%) compared with the control. FOs up regulated the mRNA expression levels of SOD, CAT, and HO-1 in organs. Moreover, the immunoblot analysis revealed increased nuclear factor-E2-related factor (Nrf2) protein expression levels in organs and there were positive correlations between the mRNA expression of phase II detoxifying/antioxidant enzymes and the expressions of Nrf2 protein, which demonstrated FOs treatment could modulate the detoxifying/antioxidant enzymes via Nrf2 signaling.

Antioxidant effect of apolipoprotein A-I on high-fat diet-induced non-alcoholic fatty liver disease in rabbits

Non-alcoholic fatty liver disease (NAFLD) is an increasingly recognized condition that encompasses a spectrum of liver abnormalities. It has been suggested that oxidative stress and lipid peroxidation are key pathophysiological mechanisms in NAFLD. Although an antioxidant effect of apolipoprotein A-I (apoA-I) has been reported, its influence on NAFLD has not been reported. The aim of this study was to determine whether apoA-I could improve the biochemical and histological abnormalities associated with high-fat diet-induced NAFLD through its antioxidant actions in rabbits. Liver damage was evaluated by hepatic coefficient, hepatic lipid assay, liver apparent abnormalities as well as hematoxylin-eosin staining of liver sections. Lipid peroxidation was assessed by measuring malondialdehyde (MDA) level in liver. Oxidative stress was assessed by measuring superoxide dismutase (SOD), glutathione peroxidase (GPx), and inducible nitric oxide synthase (iNOS) activities in serum and liver. Also, the mRNA expressions levels of SOD, GPx, and catalase (CAT) were determined by real-time quantitative polymerase chain reaction method. The results showed that apoA-I (20 or 40 mg/kg/w) was effective in reducing hepatic steatosis, inflammation, hepatic coefficient, and liver total cholesterol, triglyceride, low-density lipoprotein-cholesterol, and MDA levels in high-fat diet rabbits. In addition, apoA-I increased SOD and GPx activities while reducing iNOS activity in serum and liver. Moreover, apoA-I significantly increased the mRNA expression levels of SOD, GPx, and CAT in liver. This study showed that apoA-I exerted protective effects against fatty liver disease in rabbits induced by a high-fat diet, possibly through its antioxidant actions.

Lymphocyte Cu/ZnSOD and MnSOD Gene Expression Responses to Intensive Endurance Soccer Training

ABSTRACTThe purpose of the present study was to examine the effect of physical training on the gene expression levels of Cu/Zn superoxide dismutase (Cu/Zn SOD) and Mn superoxide dismutase (Mn SOD) in young soccer players at the beginning of their professional careers. The non-trained young men underwent a professional soccer training program of 12 weeks.Blood was obtained at two time points: before and after the 12-week endurance soccer training. Plasmatic superoxide dismutase (SOD) activity, concentration of sulfhydryl (SH) groups, superoxide anion (O2.-), malondialdehyde (MDA), and advanced oxidation protein product (AOPP) were determined. Messenger RNA (mRNA) relative levels of SOD isoenzymes were measured in peripheral blood mononuclear cells by Real-time PCR.After 12-week training, the soccer group experienced a significant decrease in plasmatic SOD activity and increase of O2.- concentration. Concentration of SH groups, MDA and AOPP were not affected by physical training, although they showed a trend to increase. Cu/Zn SOD mRNA expression levels were stable during the endurance training period but Mn SOD mRNA levels were significantly higher after the training period. The variation in Mn SOD mRNA levels was dependent on MDA concentration. Negative prediction of MDA concentration on Mn SOD mRNA expression levels remained significant after inclusion of all the other oxidative stress status parameters in a multiple regression model (p = 0.017). Positive prediction of Cu/Zn SOD mRNA levels on plasmatic SOD activity remained significant after inclusion of MDA, log O2.- and AOPP concentrations in a multiple regression model (p = 0.025).Intensive endurance soccer training stimulated a beneficial increase in lymphocyte Mn SOD rather than in Cu/Zn SOD mRNA expression levels. An interaction between the oxidative stress/antioxidative defence status parameters in blood and gene expression levels of Cu/Zn SOD and Mn SOD in PBMCs was also observed.

Effects of Soybean Isoflavones on In vitro Antioxidative Capacity of Satellite Cells of Porcine Skeletal Muscles

Abstract A synthetic isoflavone (ISO-S) or genistein was added in culture medium at different concentrations (0, 10, 20, 30, 40, and 80 μmol L −1) to investigate the effects of soybean isoflavones on antioxidative capacity of porcine skeletal muscle satellite cells. After 48 h incubation, the suspension was cryopreserved for the determination of Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) activities, and malondialdehyde (MDA) content. The mRNA levels of SOD, CAT, and GSH-Px gene in cells were detected with Taqman fluorescent probe method. The results showed that the content of MDA and the activities and the mRNA levels of SOD of porcine skeletal muscle satellite cells were influenced by supplemented soybean isoflavone (P<0.05) when adding 10-80 μmol L −1 ISO-S or genistein in the medium. The MDA contents, SOD and CAT activities and their mRNA expression levels of porcine skeletal muscle cells responded quadratically ( P <0.05) as the level of ISO-S or genistein increased. Pre-incubation of porcine skeletal muscle satellite cells with ISO-S or genistein at 10-40 μmol L −1 elevated the activities and the mRNA expression levels of SOD and CAT in cells concurrently and decreased the cellular content of MDA (P<0.05). The results indicated that pre-incubation of ISO-S or genistein at 10-40 μmol L −1 could improve the antioxidative capacity of porcine skeletal muscle satellite cells.

The effects of acute acetaminophen toxicity on hepatic mRNA expression of SOD, CAT, GSH-Px, and levels of peroxynitrite, nitric oxide, reduced glutathione, and malondialdehyde in rabbit

We investigated the regulation of antioxidant system under acetaminophen (AAP) toxicity. Twelve male New Zealand rabbits were divided into two groups with the following treatments: Group 1 animals were intraperitoneally injected with single saline (control). Group 2 animals were treated with intraperitoneal injection of AAP at a dose of 250 mg/kg body weight. Four hours following the treatments, blood samples were collected and the rabbits were sacrificed to collect liver samples. Hepatocellular damage was evaluated by aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels as well as histopathological examinations and immunohistochemical analysis. Tissue-reduced glutathione (GSH), nitric oxide (NO(.)), and malondialdehyde (MDA) levels were also measured. mRNA expression levels of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) were measured by semi-quantitative RT-PCR. It was found that liver GSH was reduced significantly in AAP-treated rabbits (P < 0.05), while MDA and NO(.) levels were increased when they were compared to control (P < 0.05). Blood AST and ALT levels were also increased following AAP treatment (P < 0.05). Hepatocellular degeneration and severe necrosis were detected in histopathological examinations. Increased immunostaining was observed for inducible nitric oxide synthase (iNOS) and nitrotyrosine in the liver. There were no changes in mRNA expression levels of SOD, CAT, and GSH-Px after AAP treatment compared to control group. These results suggest that the expression of these enzymes, which are involved in the antioxidant system, may not be altered after AAP toxicity, although classical toxic changes such as depletion of GSH, hepatocellular necrosis, and increased immunostaining for iNOS and nitrotyrosine were detected.

The effect of quercetin on the mRNA expression of different antioxidant enzymes in hepatoma cells.

The flavonol quercetin shows a wide range of effects in biological systems. We investigated whether quercetin exerts its proposed antioxidant properties via the antioxidant enzyme system. Quercetin in a concentration range from 5 to 100 microM decreased manganese superoxide dismutase, glutathione peroxidase, and copper zinc superoxide dismutase mRNA expression levels each by 30-40% in rat hepatoma H4IIE cells. Catalase mRNA expression levels increased about 30% but only with the cytotoxic concentration of 100 microM. Despite the down-regulation of antioxidant enzyme mRNA expression quercetin treatment of cells induced only a mild oxidative stress. Pretreatment of H4IIE cells with quercetin even protected against an oxidative stress resulting from hydrogen peroxide exposure. In conclusion, the antioxidant capacity of quercetin was shown not to be due to the antioxidant enzyme system.