Catalase mRNA Research Articles

EARLY EFFECTS OF EXTRACELLULAR VESICLES SECRETED BY ADIPOSE TISSUE MESENCHYMAL CELLS IN RENAL ISCHEMIA FOLLOWED BY REPERFUSION: MECHANISMS RELY IN THE RESTORATION OF THE REDOX TISSULAR ENVIRONMENT

<h3>Background</h3> The acute kidney injury (AKI) caused by ischemia followed by reperfusion (I/R) is one of the more frequent and severe processes, which, with very high mortality, affects renal structures and function. The first hours of lesion are critical for organ recovery, but the mechanisms of lesions to support early beneficial therapeutic actions are not well understood. AKI is characterized by intense production of anion superoxide (O2•-) and oxidative damage. In this study we investigated whether extracellular vesicles secreted by adipose tissue mesenchymal cells (EVs) administrated at the moment of reperfusion, suppresses the exacerbated mitochondrial O2•- formation in the first 24 h after I/R. <h3>Methods</h3> Rats were submitted to bilateral renal arterial clamping (30 min) followed by 24 h of reperfusion. The animals received subcapsularly EVs (I/R+EVs group) or saline (I/R group). The 3rd group was of the false-operated rats (SHAM). Mitochondria were isolated from proximal tubule cells and immediately used. EVs were characterized by cytometry (CD9, TSG101, CD63, CD81) and transmission electron microscopy. The probe Amplex Red™ was used to measure mitochondrial O2•- formation and MitoTracker® Orange to evaluate the mitochondrial electrical potential (Δψ). In vitro studies were carried out by using human renal cells (HK-2) co-cultured or not with EVs under hypoxia conditions. <h3>Results</h3> Administration of EVs to I/R rats restored O2•- formation to SHAM levels in all mitochondrial functional conditions: non-phosphorylating, phosphorylating, with ATP synthesis blocked and uncoupled. Proton leak was not modified. During this period, AKI remained active, as demonstrated by qRT-PCR analysis of the lesion biomarkers Kidney Injury Molecule-1 and Neutrophil Gelatinase-Associated Lipocaline. Tubular catalase and superoxide dismutase mRNA remained low. Heme oxygenase-1 (HO-1) was intensely upregulated. The co-cultures of HK-2 cells with EVs revealed decrease of apoptosis. <h3>Conclusions</h3> These findings provide evidence that the mechanisms in which rely the early recovery by EVs of the renal structure and function after I/R are those related with the normalization of the mitochondrial redox environment and open new vistas for the acellular treatment of AKI.

Effects of fish meal under different storage conditions on growth, serum biochemical indices and antioxidant capacity for juvenile grouper Epinephelus coioides

This research aimed to assess effects of fish meal under different storage conditions on growth and antioxidant capacity for juvenile grouper within eight weeks. The healthy fish samples (initial body weight = 18.75 ± 0.05 g) were irregularly distributed to 0.5 m3 barrels with 28 fish per barrel, fed with each diet in the three replicate groups. Fish meals (FMs) were stored at 4°C (experiment 1) and room temperature (30 ± 2°C, experiment 2) for 45, 90 and 135 days, and the data of VBN, HA, AV and TBA were detected. Seven iso-nitrogenous and iso-lipidic diets were prepared and denoted as fresh fish meal FFM (control group), 4-FM45, 4-FM90, 4-FM135, RT-FM45, RT-FM90 and RT-FM135, respectively. Prolonged storage time, the volatile base nitrogen, acid value, histamine and thiobarbituric acid contents of FM increased significantly (p < .05). The growth condition and feed utilization of experimental fish were not significantly affected (p > .05). Prolonged storage time, in experiment 1, the catalase (CAT), the superoxide dismutase (SOD) activities and the malondialdehyde (MDA) contents initially increased dramatically and then decreased (p < .05). The reactivity of CAT mRNA in FFM group was significantly higher than that in other treatment groups (p < .05). In experiment 2, CAT activity decreased significantly (p < .05), whereas the MDA contents increased sharply (p < .05). SOD activity and CAT expression in the liver initially increased and then decreased (p < .05). The mRNA level and enzyme activity of trypsin decreased (p < .05) in experiment 1 and 2. In summary, storage of FM (within 135 days) cannot improve the growth and feed utilization of juvenile grouper but weaken the antioxidant capacity of the liver.

Ascorbic acid coadministration with artesunate\u2013amodiaquine, up-regulated antioxidant enzymes gene expression in bone marrow cells and elicited biochemical changes in Plasmodium berghei-infected mice

One of the hallmarks of malaria infection is oxidative stress. This study was aimed at investigating the potential effects of coadministering a therapeutic dose of artesunate–amodiaquine (AS/AQ) with Vitamin C (Vit C) orally on some biochemical parameters and antioxidant enzymes gene expression in bone marrow cells of Plasmodium berghei-infected mice. Thirty male Swiss albino mice were divided into five groups of six mice each as follows: Basal control (not infected with P. berghei), Untreated (P. berghei-infected without treatment), Vit C, AS/AQ and AS/AQ + Vit C combination treated mice. Treatment was done twice daily for three consecutive days. Complete parasite clearance was observed on the second day of treatment in AS/AQ and AS/AQ + Vit C combination treated P. berghei-infected mice. Serum albumin and bilirubin levels were higher in the AS/AQ + Vit C combination treated P. berghei-infected mice compared with those treated with AS/AQ only. Artesunate–amodiaquine + Vit C combination increased superoxide dismutase activity and reduced hydrogen peroxide and malondialdehyde levels in P. berghei-infected mice when compared with the mice treated with only AS/AQ. Furthermore, AS/AQ + Vit C combination significantly up-regulated catalase and glutathione peroxidase-1 (GPx-1) mRNA expression compared with the mice treated with only AS/AQ. This is the first report linking AS/AQ to antioxidant enzyme gene expression in bone marrow cells. Our findings showed that AS/AQ and Vit C coadministration may be beneficial as it ameliorated oxidative stress and up-regulated antioxidant enzyme gene expression in P. berghei-infected mice.

Effect of Lactobacillus fermentum HFY03 on the Antifatigue and Antioxidation Ability of Running Exhausted Mice.

Yak yogurt is mainly produced in Qinghai-Tibet Plateau. It is a kind of naturally fermented dairy product. It contains abundant microorganisms. Lactobacillus fermentum (LF) HFY03 is a lactic acid bacteria derived from it. Our main research content is to study the influence of LF-HFY03 on the antifatigue and antioxidation ability of running exhausted mice. We gave different doses of LF-HFY03 to mice by gavage for 4 weeks. We selected vitamin C as the positive control group, mainly to study the relationship between antioxidant capacity and fatigue resistance and LF-HFY03 in mice with running exhaustion. The results showed that LF-HFY03 and vitamin C could significantly improve the running time of mice. And with the increase in LF-HFY03 concentration, the exhaustion time of mice was also extended. LF-HFY03 can reduce the content of urea nitrogen and lactic acid and also can increase the content of free fatty acids and liver glycogen. The levels of alanine aminotransferase, serum creatine kinase, and aspartate aminotransferase in mice decreased gradually as the antioxidant peptide level of walnut albumin increased. LF-HFY03 can reduce malondialdehyde (MDA) levels in a quantification-dependent manner and can also increase catalase (CAT) and superoxide dismutase (SOD) levels. LF-HFY03 can also increase the expressions of CAT mRNA, Cu/Zn-SOD, and Mn-SOD in the liver of mice. At the same time, LF-HFY03 can also increase the expression of protein of threonine transporter 1 (AST1)/alanine/cysteine/serine, mRNA, nNOS, and eNOS. At the same time, the solution could reduce the expression of TNF-α, syncytin-1, and inducible nitric oxide synthase (iNOS). The results showed that LF-HFY03 has a high development and application prospect as an antifatigue probiotic nutritional supplement.

Thermal Processing has no Impact on Chemopreventive Effects of Oat and Barley Kernels in LT97 Colon Adenoma Cells

The unique dietary fiber composition with high contents of β-glucan contributes to the health-promoting properties of oat and barley and may mediate a reduction of colon cancer risk. In the present study, chemopreventive effects of oat and barley (beta®barley) kernels were investigated. In order to address the impact of thermal processing on these effects, kernels were roasted (150–180 °C, approx. 20 min), digested and fermented using an In Vitro human digestion model. Concentrations of short-chain fatty acids (SCFA) and ammonia were determined in fermentation supernatants (FS). Growth inhibition, apoptosis, DNA integrity and gene expression of catalase were analyzed in LT97 colon adenoma cells. Concentrations of SCFA, particularly butyrate, were higher in oat/barley FS (2.2-fold, on average), while ammonia levels were significantly lower (0.7-fold, on average) than in the fermentation control. Treatment of LT97 cells with FS of oat/barley kernels led to a significant time- and dose-dependent growth reduction, a significant increase in caspase-3 activity and enhanced levels of catalase mRNA, without exhibiting genotoxic effects. In general, the results indicate a chemopreventive potential of In Vitro fermented oat and waxy winter barley mediated mainly by growth inhibitory and apoptotic effects, which are preserved after thermal processing.

Molecular cloning, the characterization of metallothionein and catalase, and the evaluation of testicular toxicity of Cd in the Chinese fire-bellied newt (Cynops orientalis)

Cadmium (Cd) is an environmental toxicant and a nonessential metal. Cd can attack a wide range of organs, such as the liver, kidney, lung, ovary, testis, brain, and muscle in vertebrates. Among these organs, the testis might be the most sensitive organ to Cd toxicity. Metallothionein (MT) is a cysteine-rich protein with a low molecular weight, that can bind with Cd and eliminate reactive oxygen species (ROSs). Hydrogen peroxide, which as a crucial type of ROS that is induced by Cd, can be eliminated by catalase (CAT) in the self-protection of cells and to realize Cd toxicity resistance. To investigate the functions of MT and CAT in the testis of Cynops orientalis, we cloned the full-length MT and CAT genes of C. orientalis for the first time. Immunofluorescence results demonstrated that MT and CAT were expressed in Sertoli cells and all spermatogenic cells in the testis of C. orientalis. The results of the ultrastructural damage assay demonstrated that there were various impairments, which included organelle vacuolization, abnormal chromatin distribution, and apoptotic bodies, in somatic cells that were exposed to Cd. However, the anomalies of spermatozoa were located mainly in the mid-piece and head, many of which showed severely impaired structures. The results demonstrated that MT and CAT expression had distinct patterns in response to various Cd concentrations: an increase in MT mRNA levels with elevated Cd levels and a persistent increase in CAT mRNA levels with elevated Cd levels. These results suggested that MT and CAT play roles in Cd toxicity resistance in the testis and that the expression of CAT may be a better biomarker than the expression of MT for assessing Cd pollution.

Effect of lemon peel flavonoids on anti-fatigue and anti-oxidation capacities of exhaustive exercise mice

In this study, lemon peel flavonoids (LPF) were administered to investigate its effect on the anti-fatigue and antioxidant capacity of mice that undergo exercise until exhaustion. LPF (88.36 min in LPFH group mice) significantly increased the exhaustion swimming time compare to the untreated mice (40.36 min), increased the liver glycogen and free fatty acid content in mice and reduce lactic acid and BUN content in a dose-dependent manner. As the concentration of lemon peel flavonoids increased, the serum creatine kinase, aspartate aminotransferase, and alanine aminotransferase levels of mice gradually decreased. LPF increases superoxide dismutase (SOD) and catalase (CAT) levels in mice and reduces malondialdehyde levels in a dose-dependent manner. And LPF raises hepatic tissue SOD, CAT activities and reduces skeletal muscle tissue iNOS, TNF-α levels of mice compared to the control group. LPF also enhanced the expression of copper/zinc-superoxide dismutase (Cu/Zn-SOD), manganese-superoxide dismutase (Mn-SOD), and CAT mRNA in mouse liver tissue. LPF also enhanced the expression of alanine/serine/cysteine/threonine transporter 1 (ASCT1) mRNA and attenuate the expression of syncytin-1, inducible nitric oxide synthase (iNOS), and tumor necrosis factor (TNF)-α in mouse skeletal muscle. According to high-performance liquid chromatography (HPLC) analysis, it was found that LPF contains flavonoids such as rutin, astragalin, isomangiferin, naringin, and quercetin. Our experimental data show that LPF has good anti-fatigue effects and anti-oxidation ability. In summary, LPF has high prospects to be developed and added to nutritional supplements.

Effect of reproduction cycle stage on energy metabolism responses in a sentinel species (Dreissena polymorpha) exposed to cadmium: What consequences for biomonitoring?

Metal trace elements such as cadmium (Cd) are commonly present in ecosystems and could lead to impairment of mitochondrial functions and energy imbalance in aquatic organisms including molluscs. Combined exposure to increasing temperatures and Cd could enhance such an impact on animals. Seasonal fluctuations, such as temperature, and the corresponding reproduction cycle can affect biomarker responses. However, the reproduction cycle stage is rarely taken into account in ecotoxicological studies. Thus, this work aimed at understanding energy metabolism responses in a sentinel species, Dreissena polymorpha. Mussels were collected during the rest and the reproduction periods and were exposed to 10 μg.L−1 of cadmium (Cd) at two temperatures (in situ temperature and in situ temperature + 5°C) during 7 days. Energy metabolism was monitored by measuring reserves and energy nucleotides charge and by assessing aerobic and anaerobic metabolism markers, and upstream regulation pathways. Markers related to OXPHOS activity revealed seasonal variations under laboratory conditions. Conversely, adenylate nucleotides, glycogen, lipid and transcript levels of AMP-activated protein kinase, citrate synthase, ATP synthase and cytochrome b encoding genes remained steady after the acclimation period. No evident effect of Cd on energy metabolism markers was noticed for both exposures although the transcript level of succinate dehydrogenase and citrate synthase encoding genes decreased with Cd during the rest period. Cellular stress, revealed by lipid peroxidation and catalase mRNA levels, only occurred in Cd and warming co-exposed mussels during the reproduction period. These results suggest that contaminant impact might differ according to the reproduction cycle stage. The effect of confounding factors on biomarker variations should be further investigated to have a deeper knowledge of metabolism responses under laboratory conditions.

Comparative study of dietary fat: lard and sugar as a better obesity and metabolic syndrome mice model

Background Diet macronutrient heterogeneity hinders animal studies’ data extrapolation from metabolic disorders to human diseases. Objective The present study aimed to evaluate different fat-diet compositions’ effect on inducing lipid/glucose metabolism alterations in mice. Methods Swiss male mice were fed for 12 weeks with five different diets: Standard Diet (ST), American Institute of Nutrition 93 for growth (AIN93G) high-butter/high-sugar (HBHS), high-lard/high-sugar (HLHS), and high-oil/high-sugar diet (soybean oil) (HOHS). Several parameters, such as serum biochemistry, histology, and liver mRNA expression, were accessed. Results The main findings revealed that the HLHS diet dramatically altered liver metabolism inducing hepatic steatosis and increased total cholesterol, triglycerides, VLDL, increasing liver CCAAT/enhancer binding protein (CEBP-α), Acetyl-CoA carboxylase (ACC) and Catalase (CAT) mRNA expression. Moreover, the HLHS diet increased glucose intolerance and reduced insulin sensitivity. Conclusions High-fat/high-sugar diets are efficient to induce obesity and metabolic syndrome-associated alterations, and diets enriched with lard and sugar showed more effective results.

Octreotide (somatostatin analog) attenuates cardiac ischemia/reperfusion injury via activating nuclear factor (erythroid-derived 2)-like 2 (Nrf2) signaling pathway in rat model of hyperthyroidism

BackgroundHyperthyroidism is known to increase the risk of ischemic heart diseases. Octreotide has been reported to attenuate ischemia/reperfusion (I/R) injury. Whether it is useful when ischemic heart disease is accompanied with co-morbidities like hyperthyroidism needs more clarifying. So, this study aimed to explore the effect of octreotide on cardiac I/R injury in hyperthyroid rats and to clarify if Nrf2 activation is involved in this effect. Forty adult female Wistar rats were subdivided into control (euthyroid) (n = 10) and hyperthyroid (n = 30) groups. Rats in hyperthyroid group received l-thyroxine (12 mg/L) in drinking water for 35 days, then were randomly divided into three equal subgroups (n = 10): hyperthyroid control positive group, hyperthyroid octreotide treated group, and hyperthyroid octreotide + Nrf2 inhibitor (brusatol) treated group. Isolated hearts were submitted to I/R and evaluated for cardiac hemodynamics and infarct size. Serum T3 and T4, coronary efflux lactate dehydrogenase (LDH) and creatine kinase-myoglobin binding (CK-MB) and cardiac tissue malondialdehyde (MDA) were estimated. Nrf2- regulated gene expressions of HO-1, SOD, GPx, and catalase were assessed.ResultsOctreotide administration to hyperthyroid rats improved baseline and post-ischemic recovery of cardiac hemodynamics, decreased the high coronary efflux LDH and CK-MB and tissue MDA, reduced infarction size, and upregulated the decreased antioxidative enzymes HO-1, SOD, GPx, and catalase mRNA expressions in the hyperthyroid I/R rat hearts. The Nrf2 inhibitor brusatol reversed the cardioprotective effect of octreotide in hyperthyroid I/R rat hearts.ConclusionOctreotide can reduce oxidative stress to effectively alleviate I/R injury in the hyperthyroid rat hearts through upregulation of Nrf2-dependent antioxidative signaling pathways.

Catch-up growth in intrauterine growth-restricted piglets associated with the restore of pancreatic and intestinal functions via porcine glucagon-like peptide-2 microspheres

ABSTRACT Intrauterine growth restriction (IUGR) results in abnormal morphology and gastrointestinal function, such as reduced villi height and crypt depth, thinner mucosa and muscle layers, and reduced brush border enzyme activities, delayed gastric emptying, increased stress response. As a gastrointestinal growth factor, the manner by which the porcine glucagon-like peptide-2 (pGLP-2) microsphere administration restored the gastrointestinal function and growth performance of IUGR piglets was investigated. Fourteen newborn Duroc × (Yorkshire × Landrace) IUGR piglets (0.92 ± 0.113 kg) were assigned into the IUGR (negative control group) and pGLP-2 microsphere groups. The piglets in group pGLP-2 were intraperitoneally administered with 100 mg pGLP-2 microspheres on day 1 after birth. From days 15 to 26 of trial, the body weight of the pGLP-2 group was significantly higher than that of the control. IUGR piglets of group pGLP-2 showed a significantly increased pancreas weight, serum insulin content and activity of lipase and amylase. Injection of pGLP-2 microspheres restored the intestinal absorptive capacity by significantly increasing the mRNA expression of the sodium-glucose cotransporter 1 in the jejunum and the peptide transporter 1 in the jejunum. It also restored the redox balance by increasing the catalase mRNA expression and decreasing the heat shock protein 70 mRNA expression. In addition, this improvement was associated with the significant increase in gut diameter, length and weight. Therefore, it was concluded that the injection of pGLP-2 microspheres was a suitable therapeutic strategy for compensatory growth in low birth weight IUGR piglets.

Oxidative Stress and Alterations in the Antioxidative Defense System in Neuronal Cells Derived from NPC1 Patient-Specific Induced Pluripotent Stem Cells.

Oxidative stress (OS) represents a state of an imbalanced amount of reactive oxygen species (ROS) and/or a hampered efficacy of the antioxidative defense system. Cells of the central nervous system are particularly sensitive to OS, as they have a massive need of oxygen to maintain proper function. Consequently, OS represents a common pathophysiological hallmark of neurodegenerative diseases and is discussed to contribute to the neurodegeneration observed amongst others in Alzheimer’s disease and Parkinson’s disease. In this context, accumulating evidence suggests that OS is involved in the pathophysiology of Niemann-Pick type C1 disease (NPC1). NPC1, a rare hereditary neurodegenerative disease, belongs to the family of lysosomal storage disorders. A major hallmark of the disease is the accumulation of cholesterol and other glycosphingolipids in lysosomes. Several studies describe OS both in murine in vivo and in vitro NPC1 models. However, studies based on human cells are limited to NPC1 patient-derived fibroblasts. Thus, we analyzed OS in a human neuronal model based on NPC1 patient-specific induced pluripotent stem cells (iPSCs). Higher ROS levels, as determined by DCF (dichlorodihydrofluorescein) fluorescence, indicated oxidative stress in all NPC1-deficient cell lines. This finding was further supported by reduced superoxide dismutase (SOD) activity. The analysis of mRNA and protein levels of SOD1 and SOD2 did not reveal any difference between control cells and NPC1-deficient cells. Interestingly, we observed a striking decrease in catalase mRNA and protein levels in all NPC1-deficient cell lines. As catalase is a key enzyme of the cellular antioxidative defense system, we concluded that the lack of catalase contributes to the elevated ROS levels observed in NPC1-deficient cells. Thus, a restitution of a physiological catalase level may pose an intervention strategy to rescue NPC1-deficient cells from the repercussions of oxidative stress contributing to the neurodegeneration observed in NPC1.

Ethanol Extract of Chinese Hawthorn (Crataegus pinnatifida) Fruit Reduces Inflammation and Oxidative Stress in Rats with Doxorubicin-Induced Chronic Heart Failure.

BackgroundChinese hawthorn (Crataegus pinnatifida) fruit is a traditional Chinese medicine for treatment of digestive system and cardiovascular diseases. The fruit contains polyphenol compounds, such as epicatechin, that have anti-inflammatory activity. This study aimed to investigate the effects of an alcohol extract of hawthorn fruit (HAE) on inflammation and oxidative stress in rats with doxorubicin-induced chronic heart failure (CHF).Material/MethodsRats were intraperitoneally injected with doxorubicin to induce CHF and subsequently treated with HAE intragastrically once daily for 6 weeks. At the end of the experiment, echocardiographic and hemodynamic parameters were assessed, and enzyme-linked immunoassays were used to detect the levels of cardiac injury markers (brain natriuretic peptide, creatine kinase-MB, aspartate aminotransferase, lactate dehydrogenase, copeptin, and adrenomedullin), oxidative stress markers (glutathione peroxidase and malondialdehyde), and inflammatory cytokines (interleukin [IL]-6, IL-8, IL-1β, and tumor necrosis factor-α). The IL-1β, IL-6, glutathione peroxidase-1, and catalase mRNA levels were also measured by quantitative real-time polymerase chain reaction.ResultsOur findings indicated that HAE exerts a cardioprotective effect, as shown by improved echocardiographic and hemodynamic parameters, decreased activity of serum myocardial enzymes, reduced serum levels of CHF markers, and inhibited inflammatory response in cardiac tissue. In addition, HAE treatment downregulated the mRNA expression of IL-1β and tumor necrosis factor-α and upregulated the mRNA expression of glutathione peroxidase-1 and catalase compared with untreated doxorubicin-induced CHF rats.ConclusionsHAE shows promise for the prevention and treatment of CHF. The cardioprotective effect of HAE appears to be related to inhibition of both the inflammatory response and oxidative stress in vivo.

Condensed tannins enhanced antioxidant capacity and hypoxic stress survivability but not growth performance and fatty acid profile of juvenile Japanese seabass (Lateolabrax japonicus)

Condensed tannins (CT) have been shown a promising alternative to antibiotics for livestock, but little information is available about the effect of purified CT on the growth and health of fish. A 56-day feeding trial with 640 juvenile Japanese seabass (Lateolabrax japonicus) was conducted to assess the effects of CT on the growth performance, fatty acid composition, antioxidant capacity and hypoxic stress of L. japonicus. Four diets were formulated to contain 0 (CT0), 100 (CT100), 200 (CT200) and 400 (CT400) mg/kg CT isolated from grape seed, quadruplicate groups of 40 fish with initial body weight of 10.1 ± 0.11 g were fed to apparent satiation two times daily. Fish were dissected at the end of experiment to evaluate the somatic indexes. Blood samples were collected to analyze serum metabolites, antioxidant enzymes and non-specific immune indices. Muscle and hepatopancreas were sampled to determine fatty acid composition. At the end of the feeding trail, 15 fish from each tank were suffered with a 10-hr hypoxic stress. Blood and hepatopancreas of the suffered fish were collected to analyze antioxidant enzymes, and hepatic mRNA levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), catalase (CAT) and superoxide dismutase (SOD) were analyzed. Results indicated that growth performance, whole body compositions, serum biochemical parameters, and fatty acid profile in muscle and hepatopancreas of fish were not affected by dietary treatments. Intraperitoneal fat ratio of the fish was linearly decreased (P < 0.01) as the dietary CT increasing from 0 to 400 mg/kg. Serum total antioxidant capacity (TAOC) and CAT were linearly increased (P < 0.01), but tumor necrosis factor was linearly decreased (P < 0.05) and IL-6 was linearly and quadratically decreased (P < 0.001) with increasing CT. There was a linear increase (P < 0.05) in hepatic TAOC and anti-superoxide anion with increased dietary CT. Hepatic CAT was quadratically increased (P < 0.01), and GSH-Px and SOD were linearly and quadratically decreased (P < 0.05) with increasing CT. Cumulative mortality rate (CMR) after 10 h of hypoxic stress and serum MDA were linearly and quadratically decreased (P < 0.001), whereas serum CAT and GSH-Px were linearly increased (P < 0.01) and TAOC was linearly and quadratically increased (P < 0.05) with increasing dietary CT. Hepatic CAT and SOD after 10 h of hypoxic stress were linearly increased (P < 0.05) with CT increasing. The Nrf2 and HO-1 mRNA levels were up-regulated (P < 0.05) with increasing dietary CT, and CAT and SOD mRNA levels were up-regulated (P < 0.05) in CT400 than CT0-CT200. In conclusion, dietary CT up to 400 mg/kg can not only improve the antioxidant status of L. japonicus without affecting growth performance, but also reduce the CMR under hypoxic stress by activating Nrf2 signaling pathway.

Probable Protective Effect of Exogenous Administration of Growth Hormone on Age –induced Hypercholesterolemia in Rats.

Background: Aging is an inevitable biological process accompanied by many metabolic changes such as hypercholesterolemia that increased risk of cardiovascular disease in elderly. It was reported that growth hormone GH substitution contributes to dyslipidemia improvement by modulating expression of LDL R. To our knowledge, no clarified distinguished mechanisms have been puplished yet . Objectives: To test the hypothesis that GH would improve age induced hypercholesterolemia by over expression of LDL-R m RNA . Also, to distinguish its effects on the inflammatory response and the oxidative state that may clarify the mediating mechanisms. Material& method: thirty male albino rats were divided into three groups (10 rats each): adult control, aging group, GH aged treated groups. At the end of experiment, retro-orbital blood samples were taken for estimation, lipid parameters and serum amyloid a then liver were removed for measurement of tissue catalase and LDL-R mRNA real time. Results: The present study revealed that aging worsened lipid profile. Fortunately GH substitution leads to lipid profile improvement. These metabolic changes might assert to oxidative state and inflammatory response improvement accompanying apparent LDL-R mRNA real time. Conclusion: GH supplementation can positively modulate the lipid profile in aging through modifying effect on the inflammatory response , anti-oxidant state and over expression of hepatic LDL-R . The clinical inferences of these results require further investigations.

Abstract 537: Combination of Mitochondrial and Cytosolic Antioxidant Upregulated MSC Therapy Improves Non-Alcoholic Fatty Liver Disease (NAFLD)

Introduction: Diabetes and obesity are known to cause adipose inflammation, elevated oxidative stress by reactive oxygen species (ROS) accumulation. In this report we studied the effect of human adipose derived mesenchymal stroma cells (MSCs) overexpressing antioxidants both mitochondrial (Sod2) and cytosolic (Catalase) to reduce oxidative stress and systemic inflammation in DIO mouse model. Methods: C57BL/6J male mice (4–6 weeks old) were obtained from the Jackson Lab. Obesity, glucose intolerance, and insulin resistance were induced by feeding the mice a 60% fat high-fat diet (HFD) for 12 weeks. Mouse adipose-derived MSCs and adenovirus constructs containing GFP (Null, SOD2 and Catalase), were obtained. Liver and fat tissues gene expression, liver and fat depots H&amp; E staining and liver triglycerides (L-TG) were quantified. Results: Triglyceride assay confirmed reduced liver fat accumulation in animals that received combination -MSCs. The gene expression analysis by qPCR for liver and omental fat shows upregulation of Ucp1, Pgc1a and Prdm16 genes with concomitant increase in all antioxidant (Sod2, Sod3 and Catalase) mRNA expression. Further the systemic TNF-α levels were decreased in the mice received MSC-Sod2+catalase. Glucose tolerance showed a improvement trend with low area under curve (AUC) values for mice that received MSCs with both Sod2 and catalase upregulated compared to control null group. Discussion: The combination effect of both mitochondrial and cytosolic anti-oxidants reduced systemic inflammation and liver triglyceride levels in diet induced obesity mouse model compared to SOD2, Catalase and Null MSCs. An increase mRNA expression of genes associated with browning of white adipose tissue deposits might have contributed to improvement in NAFLD along with systemic reduction in TNF-α. Genetically modified MSC therapy can be a promising option for treating obesity and diabetes.

Effect of in vitro sodium fluoride treatment on CAT, SOD and Nrf mRNA expression and immunolocalisation in chicken (Gallus domesticus) embryonic gonads

In this study, we examined the effect of sodium fluoride (NaF) on oxidative stress in chicken embryonic gonads. Following exposure to varying concentrations of NaF for 6 h, mRNA expression and immunolocalisation of catalase (CAT), sodium dismutase (SOD1 and SOD2) and nuclear respiratory factors (Nrf1 and Nrf) were analysed in the gonads. In the ovary, a dose-dependent increase in mRNA expression of CAT, Nrf1 and Nrf2 following NaF exposure was found, while the intensity of immunolocalised CAT, SOD2 and Nrf1 was higher in NaF-treated groups. In the testis, no effect of NaF on CAT, SOD1 and Nrf1 mRNA levels was observed; however, NaF (3.5–14.2 mM) elevated Nrf2 mRNA expression. NaF, at a dose of 7.1 mM, increased the immunoreactivity of Nrf1 and SOD2. Further experiments evaluated the ovary and testes when incubated with NaF (7.1 mM), vitamin C (Vitamin C, 4 mM) or NaF + Vitamin C. mRNA expression of all four examined genes in the whole ovary and immunoreactivity of Nrf1 and CAT in the ovarian medulla increased in each experimental group. Similar effects were observed in the testis, where mRNA expression, as well as CAT and Nrf2 immunoreactivity, increased in Vitamin C and NaF + Vitamin C-treated groups. In summary, NaF exposure generated oxidative stress which is manifested by increased expression of free radical scavenging enzymes in chicken embryonic gonads. High doses of Vitamin C did not reverse this effect.

Lipid-lowering effect and mechanism of Danhong Injection on hyperlipidemia rats based on lipid metabolism disorder

The animal model of hyperlipidemia in rats was established to investigate the lipid-lowering effect and mechanism of Danhong Injection on hyperlipidemic rats. SD rats were selected as the research object. The rats in normal group were fed with basic diet, and the rats in other groups were fed with high-fat diet to establish hyperlipidemia model. The successfully modeled rats were randomly divided into model group, Danhong Injection low, medium, high dose(1.0, 2.0, 4.0 mL·kg~(-1)) groups, and simvastatin(2.0 mg·kg~(-1)) group. Danhong Injection groups received intraperitoneal administration, and simvastatin group received intragastrical administration, once a day for 4 weeks. At the first, second, third, and fourth weekends after administration, blood was collected from the orbital vein to detect the levels of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C), and then the atherosclerosis index(AI) was calculated. After 4 weeks of administration, the animals were sacrificed, and their heart, liver, spleen, lung, kidney and adipose tissue were extracted and weighed respectively to calculate the organ index of each group. The expressions of acyl-coaoxidase 1(Acox1), adenosine 5'-monophosphate(AMP)-activated protein kinase alpha(AMPK-α), bile salt export pump(BSEP), peroxisome proliferator-activated receptor gamma(PPAR-γ), catalase(CAT) and superoxide dismutase(SOD) mRNA in liver tissues were detected by fluorescence quantitative PCR; the content of cholesteryl ester transfer protein(CETP) and lecithin cholesterol acyltransferase(LCAT) in serum was detected by ELISA. The results showed that as compared with the normal group, the levels of serum TC, TG and LDL-C in the model group were significantly increased, and the level of HDL-C was significantly decreased, indicating that the hyperlipidemia rat model was successfully constructed. As compared with the model group, Danhong Injection could decrease the contents of TC, TG, LDL-C and increase the content of HDL-C in hyperlipidemia rats; reduce the body weight of hyperlipidemia rats, and reduce the liver weight, liver index, fat weight and fat index; it had no significant effect on the main organ indexes such as heart, spleen, lung and kidney; but it could increase the expressions of Acox1, AMPK-α, BSEP, PPAR-γ, CAT and SOD mRNA in liver tissues of rats; it could also reduce the level of CETP and increase the level of LCAT in serum; and the regulatory effect of Danhong Injection groups all showed a dose-dependent effect. It can be concluded that Danhong Injection can regulate the blood lipid contents, reduce the blood lipid levels and alleviate the accumulation of body fat in rats with hyperlipidemia. The mechanism may be related to inhibiting lipid metabolism disorder and oxidative stress induced by high-fat diet feeding, and improving the imbalance of lipid transport system.

Protective Effects of Echium amoenum on Oxidative Stress and Gene Expression Induced by Permethrin in Wistar Rats

Background: Echium amoenum (E. amoenum), as one of the most popular plants in Iran, is traditionally used to treat different types of disorders. Objectives: This experimental study aimed to evaluate the modulatory effects of E. amoenum on permethrin (PMN)-induced oxidative stress in rats and to determine the cytoprotective effect of E. amoenum on PMN in SK-Hep-1 cells. Methods: Twenty-four male Wistar rats were randomly divided into four equal groups, including the control (normal saline), orally treated PMN (125 mg/kg of PMN), E. amoenum (100 mg/kg), and E. amoenum + PMN groups for 28 days. The levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), lipid peroxidation (LPO), catalase (CAT), and glutathione peroxidase (GPx), as well as the expression of catalase (CAT) and glutathione peroxidase (GPx), were measured in the liver of all rats. Also, the cytoprotective effect of E. amoenum against PMN was evaluated in the treated SK-Hep-1 cells. Results: The results indicated that LPO increased significantly in the PMN-treated group, as evidenced by the high concentration of malondialdehyde (MDA) in the liver. Alterations of the antioxidant system were also confirmed by the significant decline in CAT and GPx activities (2.9 ± 0.14 and 0.5 ± 0.03, respectively; P &lt; 0.05) and the significant downregulation of CAT (0.4 ± 0.02 folds) and GPx (0.3 ± 0.01 folds) mRNA expression in the liver (P &lt; 0.05). PMN also stimulated significant changes in hepatic biomarkers and induced pathological changes in the liver. On the other hand, administration of E. amoenum significantly reduced abnormalities in biochemical markers, LPO, antioxidant enzymes, gene expression, and pathological complications induced by PMN (P &lt; 0.05). E. amoenum also exhibited cytoprotective effects against cytotoxicity induced by PMN in SK-Hep-1 cells. Conclusions: The present results demonstrated that E. amoenum has significant antioxidant, gene-regulating, and cytoprotective effects.

Effects of diets high in corn oil or in extra virgin olive oil on oxidative stress in an experimental model of breast cancer.

Experimental evidence highlights the importance of dietetic factors on breast cancer. In this work we aimed to analyze the effects two oils, corn oil (rich in n-6 polyunsaturated fatty acids -PUFA-) and extra virgin olive oil (EVOO), on oxidative stress in an animal model of breast carcinogenesis. Female rats were fed a low-fat control, a high-corn oil, or a high-EVOO diet from weaning or after induction with 7,12-dimethylbenz[a]anthracene at 53 days. Animals were euthanized at 36, 51, 100 and 246 days of age. We analyzed antioxidant enzymes (mRNA and activity of superoxide dismutase, glutathione peroxidase and catalase), non-enzymatic capacity (oxidized and reduced glutathione) and DNA damage (8-oxo-dG) in tumors and mammary gland at different ages. We also analyzed lipid peroxidation (isoprostanes in serum and lipofuscin in liver). Results indicated a decrease in the enzymatic antioxidant capacity and increased oxidative stress in mammary gland of healthy young animals after a short period of high-fat diets intake, followed by an adaptation to chronic dietary intervention. After induction both diets, especially the one high in n-6 PUFA, increased the oxidized glutathione. In tumors no clear effects of the high-fat diets were observed, although in the long-term lipofuscin and 8-oxo-dG suggested greater oxidative damage by effect of the n-6 PUFA-rich diet. Considering the differential effects of these diets on mammary carcinogenesis that we have previously reported, this study suggests that these high-fat diets could have an effect on oxidative stress that would lead to different signaling pathways.