Superoxide Dismutase Gene Expression Research Articles

Antioxidant gene expression and metabolic responses of earthworms (Eisenia fetida) after exposure to various concentrations of hexabromocyclododecane

Hexabromocyclododecane (HBCD), a ubiquitous suspected contaminant, is one of the world's most prominent brominated flame retardants (BFRs). In the present study, earthworms (Eisenia fetida) were exposed to HBCD. The expression of selected antioxidant enzyme genes was measured, and the metabolic responses were assessed using nuclear magnetic resonance (NMR) to identify the molecular mechanism of the antioxidant stress reaction and the metabolic reactions of earthworms to HBCD. A significant up-regulation (p < 0.05) of superoxide dismutase (SOD) gene expression was detected, with the highest gene expression level of SOD appearing at a dose of 400 mg kg−1 dw (2.06-fold, p < 0.01). However, the glutathione transferase (GST) gene expression levels did not differ significantly (p > 0.05). Principal component analysis (PCA) of the metabolic responses showed that all groups could be clearly differentiated, and the highest concentration dose group was the most distant from the control group. Except for fumarate, the measured metabolites, which included adenosine triphosphate (ATP), valine, lysine, glycine, betaine and lactate, revealed significant (p < 0.05) increases after 14 days of exposure to HBCD. HBCD likely induces high levels of anaerobic respiration, which would result in high levels of ATP and lead to the disintegration of proteins into amino acids, including valine and lysine, to produce energy. The observed changes in osmotic pressure were indicative of damage to the membrane structure. Furthermore, this study showed that NMR-based metabolomics was a more sensitive tool than measuring the gene expression levels for elucidating the mode of toxicity of HBCD in earthworm exposure studies.

Manganese enhances the expression of the manganese superoxide dismutase in cultured primary chick embryonic myocardial cells

In the present study, the effect of manganese (Mn) on antioxidant status and the expression of the manganese superoxide dismutase (MnSOD) gene in cultured primary myocardial cells collected from the chick embryos was investigated. The hypothesis that Mn supplementation would enhance the expression of MnSOD in cultured primary myocardial cells of chick embryos was tested. Eggs collected from Mn-depleted Arbor Acres laying breeder hens were incubated for 10 days and then myocardial cells were isolated and cultivated for 8 days. The embryonic myocardial cells on day 6 were treated with Mn in the cell culture medium at different time points when the proportion of cells showing spontaneous contraction was over 95% after the 3-day primary culture. A completely randomized design involving a 3 Mn levels (0, 0.5 and 1.0 mmol L−1)×3 incubation time points (12, 24 and 48 h) factorial arrangement of treatments (n=6) was used in the current experiment. The results showed that MnSOD activity and mRNA expression level were induced by Mn and increased with incubation time, which supported the hypothesis that Mn would enhance the expression of the MnSOD gene, and thus might protect myocardial cells from oxidative stress during the chick embryonic development.

Curcumin-supplemented diets improve antioxidant enzymes and alter acetylcholinesterase genes expression level in Drosophila melanogaster model.

Curcumin, a bioactive polyphenoliccompound in turmeric (Curcuma longa) rhizomes has been shown to exert anti-aging properties with limited scientific basis. Hence, this study sought to examine the antioxidant and anti-cholinesterase activities of curcumin-supplemented diets as well as their molecular effect on superoxide dismutase (SOD) and acetylcholinesterase (AChE) genes expression level associated with lifespan extension in Drosophila melanogaster model. In this experiment, D. melanogaster (both genders) of 1 to 3days old were fed diets either containing no curcumin (control) or supplemented with curcumin at 0.2 and 1.0mg/g of diet for 7days. Subsequently, the survival and locomotor activities were determined. In addition, we evaluated RT-PCR expressions of SOD and AChE mRNA genes. Furthermore, catalase, SOD and AChE activities were determined. Curcumin-supplemented diet improves survival ability but did not affect locomotor activity when compared with the control. In addition, there was a significant increase in SOD and catalase with a concomitant decrease of AChE activities when compared with the control. Furthermore, curcumin-supplemented diets suppress AChE mRNA expression but no alteration on SOD gene expression level was observed when compared with control. In conclusion, our present results suggest that a down-regulation of AChE gene expression with a concomitant decrease of AChE activity as well as improving antioxidant status could be some possible mechanism in which curcumin exert anti-aging potential and increases lifespan of D. melanogaster.

Chitosan Controls Postharvest Decay and Elicits Defense Response in Kiwifruit

Chitosan is an eco-friendly alternative to synthetic fungicides for managing postharvest decay of fruits and vegetables. The ability of chitosan to enhance resistance to postharvest gray (Botrytis cinerea) and blue mold (Penicillium expansum) in kiwifruit was investigated. The gene expression and activity of antioxidant enzymes, and total phenolic compounds were determined. Results showed that chitosan at 5 g/L significantly inhibited gray and blue mold in kiwifruit stored at 25 and 4 °C. Chitosan significantly induced the gene expression of catalase, superoxide dismutase, and ascorbate peroxidase, as well as increased the enzyme activity. Moreover, chitosan markedly increased the content of total phenolic compounds in kiwifruit. Importantly, chitosan also exhibited beneficial effects on fruit quality. Taken together, the ability of chitosan to reduce mold in stored kiwifruit may be associated with the elicitation of host defense response. These results have practical implications for the application of chitosan to reduce postharvest losses.

Abstract LB-168: Differential response to genistein on prostate cancer cells expressing different MnSOD variants

Abstract Soy consumption may prevent prostate cancer (PCa) and be beneficial for patients with this malignancy. However, there are contradictory data on the anticancer effects of the soy isoflavone genistein, which has antioxidant properties and can affect cell proliferation and apoptosis of cancer cells through multiple mechanisms, including estrogen receptor-β mediation. Genistein also affects the expression of the manganese superoxide dismutase (MnSOD) gene and polymorphisms in this gene are related to risk of PCa. This study addressed the hypothesis that the effects of physiological concentrations of genistein on PCa cell proliferation and apoptosis are dependent on their MnSOD genotype and ER-β status, as well as on oxidative stress levels. In addition, we determined the effects of genistein on PCa cell proliferation and apoptosis at varying input levels of reactive oxygen species (O2.-), while controlling ER-β status and MnSOD genotype. PC-3 PCa cells transfected with different MnSOD genotypes (Ala/Ala and Val/Val) were cultured and treated with genistein at physiological concentrations (0, 0.01, 0.1, 0.5, 1, 5, 10 and 50 μM). After 24, 48 and 72 hours of incubation, we determined cytotoxicity, cell proliferation, apoptosis, and ER-β expression. Cell proliferation was induced at 0,5 μM genistein in PC-3 cells with MnSOD Val genotype and decreased at 2.5 μM in Ala genotype cells at 48 hours of treatment, which suggests 30-40% lower activity of Val genotype cells compared with the Ala variant, possibly increasing susceptibility to oxidative stress. ER-β expression levels were elevated by 48 hours of genistein treatment at 0.5 μM in Val genotype cells and were decreased at 2.5 μM in Ala cells. PARP protein expression levels, determined to investigate genistein’s effect on apoptosis in these cells, started to increase at 0.5 μM in Val cells and to decrease at 2.5 μM in Ala cells. These results suggest that PC-3 cells with Val genotype have increased oxidative stress and genistein at 0.5 μM induced proliferation in these cells. By contrast, genistein decreased proliferation of PC-3 cells with Ala genotype which have higher detoxification activity of superoxide radicals. Thus, genistein causes differential responses of cells with MnSOD variants, ER-β expression, and oxidative stress potential. This study help to understand the mechanisms of genistein's effects on PCa cells under physiological conditions and assess the impact of interactions of MnSOD genotype, ER-β presence, and ROS environment in the context of non-linear dose-response relationships contrasting potential benefit with possible harm of genistein. The results of this study may have important implications for PCa patients who consume soy in therapeutic and chemopreventive settings. Citation Format: Sule Terzioglu Usak, Maarten C. Bosland, Nur Özten Kandas. Differential response to genistein on prostate cancer cells expressing different MnSOD variants [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-168. doi:10.1158/1538-7445.AM2017-LB-168

Transgenic Potato Plants Overexpressing SOD and APX Exhibit Enhanced Lignification and Starch Biosynthesis with Improved Salt Stress Tolerance

Potato (Solanum tuberosum L.) is a major crop worldwide and its productivity is severely reduced by high soil salinity. The aim of the current study was to gain insights into the response and regulation of the antioxidant system in the transgenic potato, overexpressing two genes viz. thermostable CuZn-superoxide dismutase (PaSOD) gene from polyextremophile high-altitude plant Potentilla atrosanguinea and ascorbate peroxide (RaAPX) gene from Rheum australe under salinity stress. Transgenic lines and wild type (WT) were assessed under 50, 100, and 150 mM concentrations of NaCl treatment at the anatomical, transcriptional and metabolic levels. Although superoxide dismutase (SOD) and ascorbate peroxide (APX) activities in transgenic lines were significantly higher than WT under salt stress, the secondary cell wall lignification was profoundly affected by the modulated antioxidant enzymes activities and the corresponding H2O2 levels in transgenic. Expression profiles showed that the several genes and transcription factors directly involved in lignin biosynthesis were profoundly induced in transgenic lines as compared to WT. In addition, transgenic plants harbored increased starch accumulation, improved growth attributes, and reduced accumulation of reactive oxygen species than WT plants. Together, these results indicated that the expression of SOD and APX genes in transgenic potato may function as a positive set of physiological, anatomical, and molecular adjustments in the H2O2 regulated lignin biosynthesis signaling pathway, enabling the plants to withstand severe saline conditions.

Effects of Spirulina on the functions and redox status of auditory system in senescence-accelerated prone-8 mice.

To our knowledge, the effects of Spirulina platensis water extract (SP) on hearing function have not yet been reported. This study investigated the effects of SP on the function and redox status of the auditory system. Auditory brainstem responses and redox status were compared between two groups of 3-month-old senescence-accelerated prone-8 (SAMP8) mice: the control group was fed a normal diet, and the experimental group was fed a normal diet with oral supplementation of SP for 6 weeks. Compared with the control group, the experimental group had significantly lower hearing thresholds according to auditory brainstem responses measured using click sounds and 8-kHz tone burst sound stimulation at the end of this study. The experimental group had a shorter I-III interval of auditory brainstem responses with 16-kHz tone burst stimulation than the control group that was of borderline significance. Additionally, the experimental group had significantly higher mRNA expression of the superoxide dismutase and catalase genes in the cochlea and brainstem and significantly higher mRNA expression of the glutathione peroxidase gene in the cochlea. Further, the experimental group had significantly lower malondialdehyde levels in the cochlea and brainstem than the control group. However, tumor necrosis factor–α mRNA expression was not significantly different between the control and experimental groups. SP could decrease hearing degeneration in senescence-accelerated prone-8 mice possibly by increasing superoxide dismutase, catalase, and glutathione peroxidase gene expression and decreasing damage from oxidative stress in the cochlea and brainstem.

Industrial Applications of Selected JFS Articles

Industrial Applications of Selected <i>JFS</i> Articles

Dietary Yucca schidigera extract improved growth performance and liver antioxidative function in broilers

In this study, we tested the effects of Yucca schidigera extract (YSE) on the growth performance in broilers, and furthermore, its effects on antioxidative enzyme activities and corresponding gene in the liver of broilers. A total of 128 14-day-old broiler chickens were randomly assigned to four treatments: maize-soybean meal as the basal control diet and the basal diet containing 100, 200, or 300 mg/kg of YSE, respectively, in this study. Each treatment was consisted of four replicate pens with eight broilers per pen. The experiment lasted 28 days. Average daily gain (ADG), average feed intake (AFI) and feed efficiency (FE) were recorded during grower period (d 15 to d 28) and finisher period (d 29 to d 42), respectively. On day 28 and 42, liver samples were collected to analyse superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities, malondialdehyde (MDA) concentrations, total antioxidant capacity (T-AOC), and gene expressions of SOD, CAT, GPx. The results showed that during grower period, there was no difference on growth performance, while CAT activity and its gene expression were increased at 200 mg/kg YSE level. During finisher period, 100 mg/kg YSE supplementation enhanced ADG, and 100 and 200 mg/kg YSE groups improved FE. T-AOC was improved at both 200 and 300 mg/kg. 300 mg/kg supplementation of YSE enhanced GPx and SOD activity, and decreased MDA concentration. GPx gene expression was up-regulated at 300 mg/kg level. In conclusion, YSE promoted growth performance in broilers as well as exhibited liver antioxidative ability during finisher period.

Extracellular superoxide dismutase is necessary to maintain renal blood flow during sepsis development

BackgroundExtracellular superoxide dismutase (ECSOD) protects nitric oxide (NO) bioavailability by decreasing superoxide levels and preventing peroxynitrite generation, which is important in maintaining renal blood flow and in preventing acute kidney injury. However, the profile of ECSOD expression after sepsis is not fully understood. Therefore, we intended to evaluate the content and gene expression of superoxide dismutase (SOD) isoforms in the renal artery and their relation to renal blood flow.MethodsSepsis was induced in Wistar rats by caecal ligation and perforation. Several times after sepsis induction, renal blood flow (12, 24 and 48 h); the renal arterial content of SOD isoforms, nitrotyrosine, endothelial and inducible nitric oxide synthase (e-NOS and i-NOS), and phosphorylated vasodilator-stimulated phosphoprotein (pVASP); and SOD activity (3, 6 and 12 h) were measured. The influence of a SOD inhibitor was also evaluated.ResultsAn increase in ECSOD content was associated with decreased 3-nitrotyrosine levels. These events were associated with an increase in pVASP content and maintenance of renal blood flow. Moreover, previous treatment with a SOD inhibitor increased nitrotyrosine content and reduced renal blood flow.ConclusionsECSOD appears to have a major role in decreasing peroxynitrite formation in the renal artery during the early stages of sepsis development, and its application can be important in renal blood flow control and maintenance during septic insult.

24-epibrassinolide stimulates imidacloprid detoxification by modulating the gene expression of Brassica juncea L

BackgroundPesticides cause oxidative stress to plants and their residues persist in plant parts, which are a major concern for the environment as well as human health. Brassinosteroids (BRs) are known to protect plants from abiotic stress conditions including pesticide toxicity. The present study demonstrated the effects of seed-soaking with 24-epibrassinolide (EBR) on physiological responses of 10-day old Brassica juncea seedlings grown under imidacloprid (IMI) toxicity.ResultsIn the seedlings raised from EBR-treated seeds and grown under IMI toxicity, the contents of hydrogen peroxide (H2O2) and superoxide anion (O.2−) were decreased, accompanied by enhanced activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST), guaiacol peroxidase (POD) and the content of glutathione (GSH). As compared to controls, the gene expressions of SOD, CAT, GR, POD, NADH (NADH-ubiquinone oxidoreductase), CXE (carboxylesterase), GSH-S (glutathione synthase), GSH-T (glutathione transporter-1), P450 (cytochrome P450 monooxygenase) and GST1-3,5-6 were enhanced in the seedlings raised from EBR-treated seeds and grown in IMI supplemented substratum. However, expression of RBO (respiratory burst oxidase, the gene responsible for H2O2 production) was decreased in seedlings raised from EBR treated seeds and grown under IMI toxicity. Further, the EBR seed treatment decreased IMI residues by more than 38% in B. juncea seedlings.ConclusionsThe present study revealed that EBR seed soaking can efficiently reduce oxidative stress and IMI residues by modulating the gene expression of B. juncea under IMI stress. In conclusion, exogenous EBR application can protect plants from pesticide phytotoxicity.

Effect of inulin and fulvic acid on survival, growth, immune system, and WSSV prevalence in Litopenaeus vannamei

The effect of inulin and fulvic acid, added in the feed, on growth, survival, WSSV prevalence, and immune system was studied in Litopenaeus vannamei. To the above, a bioassay, with treatments in triplicate, was performed to test different additive concentrations. WSSV analysis was done in organisms infected with a relatively high viral load using single and nested PCR. At the end of the bioassay, hemolymph was extracted and the immune system was studied in hemocytes at biochemical and genetic level (quantitative PCR). The final growth was similar in all treatments and survival was between 66,7% and 93,3%. WSSV prevalence decreased 13% as compared to control. The number of hemocytes, phenoloxidase activity, and superoxide anion concentration were similar in all treatments. Inulin and fulvic acid modulated the expression of transglutaminase, superoxide dismutase, and prophenoloxidase genes, but not the Toll receptor. Additives do not negatively affect growth of white shrimp and they protect them against WSSV when infected with a relatively high viral load. Additives did not affect the immune system effectors studied at biochemical level but they modulated the expression of some immune-related genes in L. vannamei.

Rosemary Extract-Mediated Lifespan Extension and Attenuated Oxidative Damage in Drosophila melanogaster Fed on High-Fat Diet.

Rosemary extract has a potent antioxidant activity and is widely used in the food industry. In this study, the lifespan prolonging and antioxidant activity of rosemary extract was evaluated by high-fat-induced oxidative damage in Drosophila melanogaster. The results revealed that the lifespan and climbing ability of fruit flies was enhanced significantly by feeding rosemary extract. Furthermore, feeding with rosemary extract significantly increased the enzyme activity of superoxide dismutase (SOD) and catalase (CAT), and significantly decreased the level of malonaldehyde. The gene expression of SOD, CAT, and nuclear factor erythroid-2 related factor 2 was enhanced and that for methuselah was significantly reduced. The comet assay showed that high-fat diet-induced DNA lesion was significantly reduced in larvae treated with the rosemary extract. Our results suggest that feeding with rosemary extract is effective to the extended lifespan in fruit flies by strengthening of the resistance to high-fat-induced oxidative stress and by stimulating, at least in part, the endogenous antioxidant response.

ARID1A Expression is Down-Regulated by Oxidative Stress in Endometriosis and Endometriosis-Associated Ovarian Cancer.

Oxidative stress is considered an important factor in the development of endometriosis, including its malignant transformation. Previous studies have found that AT-rich interactive domain 1A (ARID1A), a tumor suppressor gene, is frequently mutated and inactivated in endometriosis-associated ovarian cancer (EAOC), and such a change in this gene is considered an early event in malignant transformation. We observed oxidative stress status by measuring the activity of the antioxidant enzyme manganese superoxide dismutase (MnSOD), malondialdehyde (MDA), and ARID1A gene expression in tissue samples from patients with endometriosis, EAOC, or non–endometriosis-associated ovarian cancer (non-EAOC). We also induced oxidative stress in the cultured cells from patients with primary endometriosis by adding H2O2 and tested for any alteration of ARID1A gene expression based on different H2O2 concentrations. The results showed that MnSOD activity in endometriosis and EAOC was lower than in non-EAOC, but MDA levels were higher. This study also showed that oxidative stress reduced ARID1A gene expression.

Anti-aging effects of M2000 (β-D-mannuronic acid) as a novel immunosuppressive drug on the enzymatic and non-enzymatic oxidative stress parameters in an experimental model.

The anti-aging property of β-D-mannuronic acid (M2000) as a novel non-steroidal anti-inflammatory and immunosuppressive agent was investigated on several determinants relative to the oxidative stress in an animal model. Sprague-Dawley rats were used for evaluating the safety and efficacy properties of M2000 on some oxidative stress enzymes, including the following: mitochondrial superoxide dismutase (SOD2), catalase (CAT), glutathione peroxidase (GPX1), glutathione S-transferase (GST), myeloperoxidase (MPO), and inducible nitric oxide synthase (iNOS) gene expression by real-time PCR. Malondialdehyde (MDA), carbonyl protein (PCO) (the lipid and protein oxidation marker, respectively), and total antioxidant capacity (TAC) were tested in serum by biochemical analysis. In addition, cortisol as a steroid hormone was surveyed by chemiluminescence immunoassay after 12 weeks of M2000 consumption. The rats were sacrificed 3 months after daily oral administration of M2000. Our findings revealed the favorable effects of M2000 on several antioxidant enzyme and gene expression, including SOD2, CAT, GPX1, and GST; however, our results were not statistically significant. Moreover, there was no significant difference in MDA and PCO as lipid and protein oxidation markers, TAC, and cortisol compared with the control group following M2000 consumption. A slight weight increase in the M2000-treated group was also observed. Our data showed the anti-aging property of M2000 as a novel designed non-steroidal anti-inflammatory drug (NSAID) with immunosuppressive property on various oxidative stress determinants.

Honokiol induces reactive oxygen species-mediated apoptosis in Candida albicans through mitochondrial dysfunction.

ObjectiveTo investigate the effects of honokiol on induction of reactive oxygen species (ROS), antioxidant defense systems, mitochondrial dysfunction, and apoptosis in Candida albicans.MethodsTo measure ROS accumulation, 2′,7′-dichlorofluorescein diacetate fluorescence was used. Lipid peroxidation was assessed using both fluorescence staining and a thiobarbituric acid reactive substances (TBARS) assay. Protein oxidation was determined using dinitrophenylhydrazine derivatization. Antioxidant enzymatic activities were measured using commercially available detection kits. Superoxide dismutase (SOD) genes expression was measured using real time RT-PCR. To assess its antifungal abilities and effectiveness on ROS accumulation, honokiol and the SOD inhibitor N,N′-diethyldithiocarbamate (DDC) were used simultaneously. Mitochondrial dysfunction was assessed by measuring the mitochondrial membrane potential (mtΔψ). Honokiol-induced apoptosis was assessed using an Annexin V-FITC apoptosis detection kit.ResultsROS, lipid peroxidation, and protein oxidation occurred in a dose-dependent manner in C. albicans after honokiol treatment. Honokiol caused an increase in antioxidant enzymatic activity. In addition, honokiol treatment induced SOD genes expression in C. albicans cells. Moreover, addition of DDC resulted in increased endogenous ROS levels and potentiated the antifungal activity of honokiol. Mitochondrial dysfunction was confirmed by measured changes to mtΔψ. The level of apoptosis increased in a dose-dependent manner after honokiol treatment.ConclusionsCollectively, these results indicate that honokiol acts as a pro-oxidant in C. albicans. Furthermore, the SOD inhibitor DDC can be used to potentiate the activity of honokiol against C. albicans.

Debaryomyces hansenii up regulates superoxide dismutase gene expression and enhances the immune response and survival in Pacific red snapper (Lutjanus peru) leukocytes after Vibrio parahaemolyticus infection

Application of yeast is increasing to improve welfare and promotes growth in aquaculture. The halotolerant yeast Debaryomyces hansenii is normally a non-pathogenic yeast with probiotic properties and potential source of antioxidant enzymes as superoxide dismutase. Here, first, we characterized the sequence features of MnSOD and icCu/ZnSOD from Pacific red snapper, and second, we evaluated the potential antioxidant immune responses of the marine yeast Debaryomyces hansenii strain CBS004 in leukocytes which were then subjected to Vibrio parahaemolyticus infection. In silico analysis revealed that LpMnSOD consisted of 1186 bp, with an ORF of 678 bp encoding a 225 amino acid protein and LpicCu/ZnSOD consisted of 1090 bp in length with an ORF of 465 bp encoding a 154 amino acid protein. Multiple alignment analyzes revealed many conserved regions and active sites among its orthologs. In vitro assays using head-kidney and spleen leukocytes immunostimulated with D. hansenii and zymosan in response to V. parahaemolyticus infection reveled that D. hansenii strain CBS004 significantly increased transcriptions of MnSOD and icCu/ZnSOD genes. Flow cytometry assay showed that D. hansenii was able to inhibit apoptosis caused by V. parahaemolyticus in the Pacific red snapper leukocytes and enhanced the phagocytic capacity in head-kidney leukocytes. Immunological assays reveled an increased in superoxide dismutase and peroxidase activities, as well as, in nitric oxide production and reactive oxygen species production (respiratory burst) in fish stimulated with D. hansenii. Finally, our results. These results strongly support the idea that marine yeast Debaryomyces hansenii strain CBS004 can stimulate the antioxidant immune mechanism in head-kidney and spleen leukocytes.

Tocotrienol-Rich Fraction Ameliorates Antioxidant Defense Mechanisms and Improves Replicative Senescence-Associated Oxidative Stress in Human Myoblasts.

During aging, oxidative stress affects the normal function of satellite cells, with consequent regeneration defects that lead to sarcopenia. This study aimed to evaluate tocotrienol-rich fraction (TRF) modulation in reestablishing the oxidative status of myoblasts during replicative senescence and to compare the effects of TRF with other antioxidants (α-tocopherol (ATF) and N-acetyl-cysteine (NAC)). Primary human myoblasts were cultured to young, presenescent, and senescent phases. The cells were treated with antioxidants for 24 h, followed by the assessment of free radical generation, lipid peroxidation, antioxidant enzyme mRNA expression and activities, and the ratio of reduced to oxidized glutathione. Our data showed that replicative senescence increased reactive oxygen species (ROS) generation and lipid peroxidation in myoblasts. Treatment with TRF significantly diminished ROS production and decreased lipid peroxidation in senescent myoblasts. Moreover, the gene expression of superoxide dismutase (SOD2), catalase (CAT), and glutathione peroxidase (GPX1) was modulated by TRF treatment, with increased activity of superoxide dismutase and catalase and reduced glutathione peroxidase in senescent myoblasts. In comparison to ATF and NAC, TRF was more efficient in heightening the antioxidant capacity and reducing free radical insults. These results suggested that TRF is able to ameliorate antioxidant defense mechanisms and improves replicative senescence-associated oxidative stress in myoblasts.

Protective Effect of Cranberry Extracts against Oxidative Stress and DNA Damage Induced by Diclofenac Sodium in Kidney of Male Albino Rate

This work aimed to find the effect of cranberry extract (75 and 150 mg/kg·b·w) and vit. C (1 g/kg·b·w orally) on renal toxicity induced by Diclofenac sodium in male albino rats. Treated rats with diclofenac sodium with a concentration 150 mg/kg·b·w, expressed a significant increase in several parameters includes, plasma total cholesterol, LDL-cholesterol, and triglyceride as well as renal nitric oxide (NO), tumor necrosis factor-alfa (TNF-α) and TBARS. In addition, a significant reduction in renal superoxide dismutase (SOD), GSH, catalase (CAT) and plasma HDL. The present results explain that, using cranberry extract and vit. C resulted in increasing the level of GSH, CAT and SOD as well as gene expression of renal SOD, CAT and IL-22 and reduce the level of TBARs significantly which led to preventing renal tissue damage. Our results also revealed that cranberry extract can protect DNA from damage as obtained from comet essay. TM-U was elevated in DCLF treated group when compared with normal. However cranberry extract was able to reduce this elevation in dose dependant manner. Histological features in H&E taken to different groups also mirrors this findings. DCLF causes many changes in renal tissue include infiltration by inflammatory cells, attenuated glomeruli, apoptosis in tubular epithelia.

The effect of Echis coloratus venom on biochemical and molecular markers of the antioxidant capacity in human fibroblasts

ABSTRACTThis study was undertaken to examine the activities and levels of major antioxidants/oxidants in cultured human fibroblasts incubated with a sublethal dose of Echis coloratus venom (EcV).Glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR) activities and gene expression levels as well as reduced glutathione (GSH) levels, and the concurrent hydrogen peroxide (H2O2), superoxide anions (SOA), lipid peroxides (LPO) and oxidized glutathione (GSSG) generation rates were assayed in fibroblast cultures and sonicates incubated with 0.5 µg ml–1 medium EcV for 4 h at 37°C.Data indicated that the activities of all antioxidant enzymes were significantly decreased and their corresponding transcripts downregulated in EcV-incubated cells compared to controls (p < 0.001). In contrast, there were parallel equally significant increases in H2O2, SOA and LPO generation rates in venom-incubated cells compared to controls (p < 0.001). Additionally, GSH levels were significantly decreased and those of GSSG were equally significantly increased in venom-incubated cultures compared to controls (p < 0.001) leading to a lowered GSH/GSSG ratio.In conclusion, incubation of fibroblast cultures with EcV resulted in a shift towards oxidative metabolism causing severe OS. This correlated with significant downregulation in the expression levels of all investigated antioxidant genes.