Glutathione S-transferase mRNA Research Articles

Basal Expression Profile of Seven Glutathione-S-Transferases (GSTs) Genes in Six Tissues of Tilapia (Oreochromis niloticus) for Polymerase Chain Reaction Array

Study’s Excerpt The differential expression patterns of Glutathione-S-transferase (GST) isoforms in six tissues of Nile tilapia are investigated. Seven specific GST genes are examined across six tissues, and it is determined that the liver, spleen, and intestines are key sites of significant gene expression. GST isoforms could be potentially used as biomarkers for assessing xenobiotic stress in aquatic environments. Full Abstract Because aquatic habitats are being destroyed and biodiversity is declining, xenobiotic pollution of water is a critical environmental problem that has garnered a lot of attention in the past few decades. This study looked at the expression patterns of Glutathione-S-transferases isoforms (GSTMA, GSTO1LA, GSTA, GSTR1, GSTK, GSTT1, and MGST), which are genes involved in xenobiotic metabolism, in Nile tilapia. From adult Nile tilapia, six tissues—liver, spleen, intestines, gills, heart, and muscle—were chosen, and seven genes that were optimised via quantitative polymerase chain reaction (qPCR) were used for validation. Gene expressions were assessed using a PCR array that included duplicate tissues in 96-well qPCR plates, with each gene/plate being assayed six times. Water is used in the remaining wells as no template control (NTC). As reference genes, Pan Ribosomal Protein L3 (RPL3) and Pan 18S ribosomal RNA (18S RNA) were employed. The genes’ basal tissue mRNA expression was measured in each tissue. The muscle was utilised to renormalise the expression levels of all genes throughout the remaining tissues since, under assay’s circumstances the muscle showed very low signal for most of the genes. Using quantitative real-time PCR (qPCR), the result demonstrated widespread differential expression patterns of the seven GST mRNA in the liver, spleen, intestine, gills, heart, and muscle of an untreated fish. However, it was clear that the highest significant expression level of most genes (different GST’s isoforms) was in the liver, followed by the spleen and gut, then the heart and gills compared to muscle (P<0.05). The findings demonstrated widespread differential expression of the GST’s isoforms at basal level and highlighted their utility as biomarkers for xenobiotic stress in aquatic environments.

Diluted bitumen weathered under warm or cold temperatures is equally toxic to freshwater fish

Canada is one of the main petroleum producers in the world. Through its oil sands exploitation, a viscous bitumen mixed with sand, water, and clay is being produced. This bitumen is so viscous that approximatively 20%–30% of diluent needs to be added to ease transportation, resulting in a mixture called diluted bitumen (dilbit). The transport of dilbit through North America comes with a potential risk for oil spills in freshwater ecosystems at any time of the year. In this study, a mesoscale spill tank was used to study dilbit spills in freshwater to understand the effect of cold (winter-like) vs. warmer (spring- and fall-like) water temperatures on its natural weathering and their toxicity to fathead minnow (Pimephales promelas) embryos. Water samples were collected weekly during two consecutive 35-day experiments ran at either 2 or 15 °C. Each week, fish larvae were exposed for 7 days, and water analysis was performed. Chemical analysis showed that the volatile organic compound, total organic carbon, and polycyclic aromatic hydrocarbon concentrations decreased in both experiments with time, while fish larvae exposed to both temperature settings yielded increased abnormalities, EROD activity, CYP1A, and glutathione S-transferase mRNA expression levels, and decreased heart rate. Importantly, there were no major differences between the temperature regimes on dilbit weathering, highlighting that if a spill occurs in colder waters, it would be equally toxic to organisms. This work provides new data on the potential risk of oil spill for use during response planning and modelling.

Association of GSTT1, GSTM1 and GSTP1 (Ile105Val) mRNA Expression with Cardiometabolic Risk Parameters in Women with Breast Cancer and Comorbidities

Breast cancer (BC) and cardiometabolic diseases share a multifactorial and modifiable etiology, modulated by complex molecular pathways. Glutathione S-transferase (GST) plays a critical role, providing protection against xenobiotics and regulating levels of enzymes and proteins in the cell. GST variants have a significant impact on susceptibility to diseases whose pathogenesis involves oxidative stress, as is the case in many inflammatory diseases such as BC and cardiometabolic pathologies. However, the expression of these polymorphic variants has not been studied in BC. This study aimed to evaluate the presence of GST mRNA isoforms and their association with clinical and cardiometabolic parameters in women with BC. This was a case-control study, and a total of 57 participants were recruited. Concentrations of glucose and lipids in blood were measured in all the participants. GST variants (GSTT1, GSTM1 and GSTP1 Ile105Val polymorphism) were evaluated in all the participants by real-time PCR analysis. There was a significant association (p < 0.05) between the frequency of GSTP1 and LDL-c in the BC group. However, the control group showed significant associations between blood pressure with GSTT1 and GSTP1 variants with total cholesterol (TC), LDL-c, VLDL-c and triacylglycerols (TG). Therefore, GSTT1 and GSTP1 variants could be emerging biomarkers to discriminate between BC cases related or not to cardiometabolic disease factors.

Early loss of glutathione -s- transferase (GST) activity during diverse forms of acute renal tubular injury.

Glutathione‐S‐transferases (GSTs) are a diverse group of phase II detoxification enzymes which primarily evoke tissue protection via glutathione conjugation to xenobiotics and reactive oxygen species. Given their cytoprotective properties, potential changes in GST expression during AKI has pathophysiologic relevance. Hence, we evaluated total GST activity, and the mRNA responses of nine cytosolic GST isotypes (GST alpha1, kappa1, mu1/5, omega1, pi1 sigma1, theta1, zeta1 mRNAs), in five diverse mouse models of AKI (glycerol, ischemia/reperfusion; maleate, cisplatin, endotoxemia). Excepting endotoxemia, each AKI model significantly reduced GST activity (~35%) during both the AKI “initiation” (0‐4 h) and “maintenance” phases (18 or 72 h). During the AKI maintenance phase, increases in multiple GST mRNAs were observed. However, no improvement in GST activity resulted. Increased urinary GST excretion followed AKI induction. However, this could not explain the reduced renal GST activity given that it also fell in response to ex vivo renal ischemia (i.e., absent urinary excretion). GST alpha, a dominant proximal tubule GST isotype, manifested 5–10‐fold protein increases following AKI, arguing against GST proteolysis as the reason for the GST activity declines. Free fatty acids (FFAs) and lysophospholipids, which markedly accumulate during AKI, are known to bind to, and suppress, GST activity. Supporting this concept, arachidonic acid addition to renal cortical protein extracts caused rapid GST activity reductions. Based on these results, we conclude that diverse forms of AKI significantly reduce GST activity. This occurs despite increased GST transcription/translation and independent of urinary GST excretion. Injury‐induced generation of endogenous GST inhibitors, such as FFAs, appears to be a dominant cause.

Long-Term Exposure to Phenanthrene Induced Gene Expressions and Enzyme Activities of Cyprinus carpio below the Safe Concentration.

Phenanthrene (PHE) is a typical compound biomagnified in the food chain which endangers human health and generally accumulates from marine life. It has been listed as one of the 16 priority PAHs evaluated in toxicology. In order to evaluate the changes of CYP1A GST mRNA expression and EROD GST enzyme activity in carp exposed to lower than safe concentrations of PHE. Long-term exposure of carp to PHE at lower than safe concentrations for up to 25 days. The mRNA expression level and cytochrome P450 (CYP1A/EROD (7-Ethoxylesorufin O-deethylase)) and glutathione S-transferase (GST) activity were measured in carp liver and brain tissue. The results showed that PHE stress induced low-concentration induction and high-concentration inhibition of CYP1A expression and EROD enzyme activity in the liver and brain of carp. In both two organs, GST enzyme activity was also induced. However, the expression of GST mRNA was first induced and then inhibited, after the 15th day. These results indicate that long-term exposure to PHE at lower than safe concentrations still poses a potential threat to carp’s oxidase system and gene expression.

Sesame lignans upregulate glutathione S-transferase expression and downregulate microRNA-669c-3p.

Oxidative stress is associated with aging and pathologies such as cardiovascular diseases, Alzheimer’s disease, and cancer. Glutathione S-transferase (GST), a family of detoxification enzymes, plays a crucial role in countering oxidative stress. Therefore, there is a need for the development of physiologically functional foods and agricultural products, which enhance GST activity. Sesamin and episesamin are major lignans in refined sesame oil that exhibit beneficial properties including antioxidative stress effects. A previous study showed that sesamin upregulated GST activity. This study aimed to elucidate the mechanism underlying the GST activity enhancement elicited by sesame lignans. C57BL/6J mice were orally administered 20 mg/kg body weight sesame lignans (sesamin:episesamin=1:1) for 7 days. Oral administration of sesame lignans increased the GST activity in the mouse liver. Furthermore, the lignans upregulated GSTA1, GSTA4, and GSTM4 protein expression. Microarray analysis revealed that sesame lignans changed the expression of various microRNAs (miRNAs) (84 upregulated, 19 downregulated). We also found 16 miRNAs, including miR-669c-3p, that may negatively regulate GST expression among the 19 miRNAs with reduced expression caused by the sesame lignans. miR-669c is reportedly negatively correlated with GST. Additionally, we transfected NMuLi cells with an miR-669c-3p mimic and evaluated the effect of miR-669c-3p on GST mRNA and protein expressions. The results showed that the miR-669c-3p mimic suppressed the mRNA and protein levels of GSTA4 and GSTM4. In conclusion, sesame lignans increased GST protein expression and activity and downregulated miRNAs, including miR-669c-3p, which is a possible suppressor of GST.

Oxidative Stress and Changes of Important Metabolic Gene Expressions as a Potential Biomarker in the Diagnosis of Atherosclerosis in Leukocytes.

IntroductionPresenilin 1 (PSEN1), catalase (CAT), glutathione-S-transferase (GST) and paraoxonase 1 (PON1) play a vital role in prediction, diagnosis and therapy of metabolic disorders.MethodsMetabolic enzyme activities and lipid peroxidation in serum of cerebrovascular diseases (CVD) and coronary artery diseases were measured by spectrophotometric methods. mRNA was isolated from leukocytes of the patient group and healthy adult patients. Quantitative gene expression of PSEN1, CAT and GST mRNA was identified by quantitative real-time polymerase chain reaction (qPCR).ResultsThe PSEN1, CAT and GST expression in patients showed significant differences compared to the control group. PSEN1 expression in leukocytes was significantly about twice as high as that of the control group in patients with CVD. The GST, CAT and PON1 activity showed significant differences in patient groups compared to the control group.ConclusionThe mRNA expression levels can be used as a potential biomarker in the diagnosis of atherosclerosis that occurs as a result of the metabolic disorder. In atherosclerotic patients, antioxidant status is independently related to an increased risk of cardiovascular events. Antioxidant activities and mRNA expressions may have predictive value, as well as available risk factors.

Molecular Cloning and Characterization of Five Glutathione S-Transferase Genes and Promoters from Micromelalopha troglodyta (Graeser) (Lepidoptera: Notodontidae) and Their Response to Tannic Acid Stress.

Plants accumulate phenolic compounds such as tannic acid to resist insect herbivores. The survival of insects exposed to toxic secondary metabolites depends on the detoxification metabolism mediated by limited groups of glutathione S-transferases (GSTs). Micromelalopha troglodyta (Graeser) (Lepidoptera: Notodontidae) is an important foliar pest of poplar trees. GSTs play an important role in xenobiotic detoxification in M. troglodyta. Five GST genes were identified in M. troglodyta and were classified into five different cytosolic GST classes, delta, omega, sigma, theta, and zeta. Real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to determine the mRNA expression of the five cloned GSTs in the midguts and fat bodies of M. troglodyta. The mRNA expression of the five GSTs was significantly induced when M. troglodyta was exposed to tannic acid. To further understand the tannic acid regulatory cascade, the 5′-flanking promoter sequences of the five MtGSTs were isolated by genome walking methods, and the promoters were very active and induced by tannic acid. In summary, the induction of GST mRNA expression was due to the response of five MtGST promoters to tannic acid. Therefore, MtGST promoters play an important role in the regulation of GST transcription.

<p class="Body"><strong>Expression profiles of glutathione S-transferases genes in semi-engorged </strong><strong><em>Haemaphysalis longicornis</em></strong><strong> (Acari: Ixodidae) exposed to <em>Cymbopogon citratus</em> essential oil</strong></p>

Glutathione S-transferases (GSTs) are phase II detoxification enzymes, which function via combining with pesticidal molecules and catalyzing the conjugation of molecules by thiol of glutathione, so as to protect tissues from oxidative stress damage. In the tick Haemaphysalis longicornis, glutathione S-transferases (HlGST and HlGST2) have been previously identified. However, the relationship between the transcription of glutathione S-transferases and the essential oil treatment in ticks remains unexplored. Hence, in the present study, the transcription profiles of HlGST and HlGST2 mRNAs were evaluated in H. longicornis after exposure to Cymbopogon citratus essential oil. At 24 h post-exposure of H. longicornis to different sublethal concentrations of C. citratus essential oil, there was significant difference (P = 0.0001) in the transcription of HlGST. Tukey’s test showed that HlGST was significantly induced after treatment with 1% C. citratus essential oil (P = 0.0002); whereas no significant difference (P = 0.3551) was detected after treated by 2% C. citratus essential oil. No significant difference (P = 0.4555) in the transcription of HlGST2 between the treatment and the control group of 50% ethanol. Nevertheless, the down-regulation of HlGST2 in the treatment groups versus the untreated control group was not significant (P = 0.1208). The above results imply that the essential oil can modulate the expression of GST mRNA, therefore, further understanding of the underlying mechanisms of the GST at the molecular level could contribute to the development of effective control measures for ticks and tick-borne diseases.

NF-κB pathway took part in the development of apoptosis mediated by miR-15a and oxidative stress via mitochondrial pathway in ammonia-treated chicken splenic lymphocytes

Ammonia, a kind of gas with pungent smell, is harmful to livestock and people, and has bad influence on the atmosphere. However, the mechanism of splenic toxicity caused by ammonia is still poorly understood. The aim of present study was to investigate the effect of ammonia on chicken splenic lymphocytes from the perspective of apoptosis. Chicken splenic lymphocytes were divided into the control group and the two ammonium treatment groups (1 mmol/L and 5 mmol/L ammonia), and were cultured for 24 h. CCK-8, flow cytometry (FC), fluorescence microscope, quantitative real-time PCR (qRT-PCR), and Western blot were used to study the differences between different groups. The results showed that ammonia exposure increased the release of calcium (Ca)2+ and reactive oxygen species (ROS) from mitochondrion. Besides, we found an increase in mRNA levels of glutathione peroxidase (GPx), inflammation-related genes (nuclear factor-κB (NF-κB), cyclooxygenase-2 (COX-2), inducible nitric (iNOS), tumor necrosis factor-α (TNF-α), and transforming growth factor-β (TGF-β)), apoptosis-related genes (B-cell lymphoma-2 (BCL-2), Bcl-2 associated X protein (BAX), Cytochrome c (Cytc), apoptotic protease activating factor 1 (APAF1), Caspase-9, and Caspase-3), and an increase in protein levels of NF-κB, iNOS, BAX, Cytc, Caspase-9, and Caspase-3. At the same time, we found a decrease level of GPx protein expression, and a decrease level of glutathione S-transferase (GST) mRNA expression, and a decrease level of heme oxygenase-1 (HO-1) and BCL-2 mRNA and protein expression in splenic lymphocytes exposed to ammonia. Meanwhile, miR-15a expression increased under ammonia exposure. In summary, these results indicated that ammonia induced oxidative stress, promoted the release of Ca2+, Cytc, and ROS from mitochondria, and then induced mitochondria-mediated inflammatory response, finally triggered apoptosis in chicken splenic lymphocytes.

Endosulfan toxicity in Nile tilapia (Oreochromis niloticus) and the use of lycopene as an ameliorative agent

ObjectivesEndosulfan is a broad-spectrum organochlorine insecticide that has been commercially in use for decades to control insect pests and has been found to pollute the aquatic environment. The current study was carried out to investigate the toxic effects of endosulfan, an organochlorine pesticide, on Nile tilapia (Oreochromis niloticus), a freshwater fish, and the alleviating effects of lycopene on the induced toxicity. MethodsFour treatment groups of fish were investigated (3 replicates of 15 fish for each group): (1) a control group, (2) a group exposed to endosulfan, (3) a group that was fed on a basal diet supplemented with lycopene, and (4) a group that was fed on a basal diet supplemented with lycopene and exposed to endosulfan. The experiment was carried out over a 4-week period. ResultsEndosulfan negatively affected liver function, including liver enzymes and plasma proteins. Endosulfan affected blood parameters of fish and reduced the counts of red blood cells (RBCs) and white blood cells (WBCs), as well as affected immunological parameters. Endosulfan caused oxidative stress, as it decreased the values of antioxidants catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), and glutathione (GSH), and increased the level of lipid peroxide malondialdehyde (MDA). Additionally, endosulfan increased cytochrome P450 (CYP450) levels, while it decreased glutathione S-transferase (GST) mRNA transcript levels and distorted the normal histological structure of the liver, gills, and spleen of affected fish. Conversely, lycopene partially restored the aforementioned parameters when administered concomitantly with endosulfan. ConclusionThe results showed the beneficial effects of supplementing fish diets with lycopene as a natural antioxidant for ameliorating the toxicity caused by endosulfan.

Effects of stocking density on stress response, innate immune parameters, and welfare of turbot (Scophthalmus maximus)

This experiment was conducted to investigate the growth performance, stress and immune responses, and welfare of juvenile turbot (Scophthalmus maximus) under three different densities (initial density 9.3, 13.6, and 19.1 kg m−2) for 120 days in a recirculating aquaculture system. Turbot were measured every 20 days to evaluate growth biometrically and sampled every month to measure biochemical parameters and mRNA levels of some stress-related genes. No significant differences were detected in the parameters and gene expression among density groups until the final sampling, except Fulton’s condition factor, lysozyme, immunoglobulin M, and complement C3. At the end of the experiment (final density 26.11, 38.22, and 52.25 kg m−2), turbot reared in the high-density (HD) group had lower body mass increase, specific growth rate, and Fulton’s condition factor, as well as higher feed conversion ratio and coefficient of variation for weight than those reared in the low-density (LD) group (P < 0.05). Fish in the HD group had higher serum cortisol, glucose, lactate, and cholesterol levels than fish in the other groups, whereas they had lower lysozyme, immunoglobulin M, and complement C3 and C4 contents (P < 0.05). Fish reared in the HD group also had higher serum chloride and osmolality levels and higher sodium-potassium adenosine triphosphatase (Na+,K+-ATPase) activity and higher Na+,K+-ATPase gene expression levels in gills compared to the other groups (P < 0.05). The mRNA levels of cytochrome P450 1A (CYP1A) and heat-shock proteins 70 and 90 (HSP70 and HSP90) were significantly upregulated, whereas glutathione S-transferase (GST) mRNA levels were significantly downregulated in the head kidney of fish in the HD group relative to fish in the other groups at the end of this trial (P < 0.05). These results indicated that overly high stocking (~50 kg m−2) density can negatively affect the growth performance, serum biochemical parameters, osmolality levels, stress-related gene expression, and overall welfare of turbot.

Genome-wide identification and expression of the entire 52 glutathione S-transferase (GST) subfamily genes in the Cu2+-exposed marine copepods Tigriopus japonicus and Paracyclopina nana

In this study, the entire glutathione S-transferases (GSTs), the major phase II detoxification enzyme, were identified in two marine copepod species Tigriopus japonicus and Paracyclopina nana. The genome-wide identification of GSTs in T. japonicus and P. nana resulted in 32 and 20 GSTs in total, respectively. Among the identified GSTs, two specific classes of GSTs, specifically sigma and delta/epsilon GSTs were the dominant form of cytosolic GSTs in T. japonicus, while delta/epsilon and mu classes were dominant cytosolic GSTs in P. nana. In addition, Membrane-Associated Proteins in Eicosanoid and Glutathione metabolism (MAPEG) family were found in relatively higher proportion compared to other classes. Moreover, sigma, delta/epsilon, and microsomal GSTs have shown to expand through tandem duplication. To validate the detoxification function of the identified GSTs, both copepods were exposed to copper (Cu2+) and the reactive oxygen species (ROS) level and GST activity were measured. With integration of phylogenetic analysis and xenobiotic-mediated GST mRNA expression patterns along with previous enzymatic activities, the functional divergence among species-specific GST genes was clearly observed. This study covers full identification of GST classes in two marine copepod species and their important role in marine environmental ecotoxicology.

Antioxidative, anti-inflammatory and hepatoprotective effects of resveratrol on oxidative stress-induced liver damage in tilapia (Oreochromis niloticus)

Resveratrol, a dietary polyphenol, has been shown to exert antioxidation, hepatoprotection, anti-inflammation and immunostimulation. However, the effects and underlying mechanism of resveratrol on liver injury in fish are still unclear. In the present study, we investigated the potential protective effects and mechanism of resveratrol on oxidative stress-induced liver damage in tilapia. Fish were fed diet containing four doses of resveratrol (0, 0.1, 0.3, and 0.6 g/kg diet) for 60 days, and then given an intraperitoneal injection of H2O2 or saline. The results showed that administration of resveratrol significantly ameliorated H2O2-induced liver injury. In serum and liver, resveratrol treatment suppressed the oxidative stress, as evidenced by the decline of lipid peroxidation level and increase of antioxidant activity. Resveratrol also activated erythroid 2-related factor 2 (Nrf2) signaling pathway and enhanced the heme oxygenase 1 (HO-1), NAD(P) H:quinone oxidoreductase 1 (NQO-1), glutathione S-transferase (GST) mRNA levels. Meanwhile, resveratrol treatment repressed TLR2-Myd88-NF-κB signaling pathway to decrease the inflammatory response in H2O2-induced liver injury as evidenced by the lower interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and IL-8 mRNA levels and higher IL-10 mRNA level. Moreover, resveratrol treatment attenuated immunotoxicity in liver of H2O2-treated fish, accompanied by upregulation of hepcidin (HEP), complement 3 (C3) and lysozyme (LZM) mRNA levels. Overall results suggested that the protection of resveratrol on H2O2-induced liver injury, inflammation and immunotoxicity was due to its antioxidant property and its ability to modulate the Nrf2 and TLR2-Myd88-NF-κB signaling pathways.

Genome-wide identification of the entire 90 glutathione S-transferase (GST) subfamily genes in four rotifer Brachionus species and transcriptional modulation in response to endocrine disrupting chemicals

Genome-wide identification of glutathione S-transferase (GST), a major phase II detoxification enzyme, was investigated in four different aquatic model rotifer species Brachionus koreanus, B. plicatilis, B. rotundiformis, and B. calyciflorus. GSTs are ubiquitous antioxidant enzymes that play versatile function including cellular detoxification, stress alleviation, and production of the radical conjugates. Among the four rotifers, B. rotundiformis was found with the least number of GST genes (total 19 GST genes), whereas the other three species shared 23 to 24 GST genes. Among the identified GST genes, belonging to the cytosolic GST superfamily, the expansion of GST sigma classes mainly occurs through tandem duplication, resulting in tandem-arrayed gene clusters on the chromosomes. Overall, the number of genes discovered in this study was highest in the sigma class, zeta, alpha, and omega in descending order. With integration of phylogenetic analysis and xenobiotic-mediated GST mRNA expression patterns along with previous enzymatic activities, the functional divergence among species-specific GST genes was clearly observed. This study covers full identification of GST classes in three marine rotifer and one fresh-water rotifer species and their important role in marine environmental ecotoxicology.

Oxidative Stress in the Muscles of the Fish Nile Tilapia Caused by Zinc Oxide Nanoparticles and Its Modulation by Vitamins C and E.

The effects of zinc oxide nanoparticles (ZnONPs) on antioxidants in Nile tilapia muscles and the protective role of vitamins C and E were examined. Two hundred males of Nile tilapia were held in aquaria (10 fishes/aquarium). Fishes were divided into 5 groups: 40 fishes in each group; the first group was the control; the 2nd and 3rd groups were exposed to 1 and 2 mg/L of ZnONPs, respectively; and the 4th and 5th group were exposed to 1 and 2 mg/L of ZnONPs and treated with a (500 mg/kg diet) mixture of vitamin C and E mixture (250 mg/kg diet of each). Muscles were collected on the 7th and 15th day of treatments. Muscle malondialdehyde, reduced glutathione levels, superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GR), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) activities were measured after treatments. Relative quantification of SOD, CAT, GR, GPx, and GST mRNA transcripts was detected in the muscles. Results showed that MDA and GSH concentration; SOD, CAT, GR, GPx, and GST activities; and mRNA expression were significantly decreased in groups exposed to ZnONPs. Vitamins C and E significantly ameliorated the toxic effects of ZnONPs. In conclusion, vitamins C and E have the ability to ameliorate ZnONP oxidative stress toxicity in Nile tilapia.

Sulforaphane Alters β-Naphthoflavone-Induced Changes in Activity and Expression of Drug-Metabolizing Enzymes in Rat Hepatocytes.

Sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, exerts many beneficial effects on human health such as antioxidant, anti-inflammatory, and anticancer effects. The effect of SFN alone on drug-metabolizing enzymes (DMEs) has been investigated in numerous in vitro and in vivo models, but little is known about the effect of SFN in combination with cytochrome P450 (CYP) inducer. The aim of our study was to evaluate the effect of SFN on the activity and gene expression of selected DMEs in primary cultures of rat hepatocytes treated or non-treated with β-naphthoflavone (BNF), the model CYP1A inducer. In our study, SFN alone did not significantly alter the activity and expression of the studied DMEs, except for the glutathione S-transferase (GSTA1) mRNA level, which was significantly enhanced. Co-treatment of hepatocytes with SFN and BNF led to a substantial increase in sulfotransferase, aldoketoreductase 1C, carbonylreductase 1 and NAD(P)H:quinone oxidoreductase 1 activity and a marked decrease in cytochrome P450 (CYP) Cyp1a1, Cyp2b and Cyp3a4 expression in comparison to the treatment with BNF alone. Sulforaphane is able to modulate the activity and/or expression of DMEs, thus shifting the balance of carcinogen metabolism toward deactivation, which could represent an important mechanism of its chemopreventive activity.

Impairment of gill structural integrity by manganese deficiency or excess related to induction of oxidative damage, apoptosis and dysfunction of the physical barrier as regulated by NF-κB, caspase and Nrf2 signaling in fish

This study is for the first time to explore the possible effects of dietary manganese (Mn) on structural integrity and the related signaling in the gills of fish. Grass carp (Ctenopharyngodon idella) were fed with six diets containing graded levels of Mn [3.65–27.86 mg Mn/kg diet] for 8 weeks. The results firstly demonstrated that Mn deficiency aggravated inflammation indicated by up-regulation of pro-inflammatory cytokines (tumour necrosis factor α, interleukin 8, and interleukin 1β mRNA levels) and down-regulation of anti-inflammatory cytokines (interleukin 10, transforming growth factor-β1) mRNA levels, which might be partially related to the up-regulation of nuclear factor kappa B (NF-κB p65) and down-regulation of nuclear inhibitor factor κBα (iκBα) mRNA levels in the gills of fish. Meanwhile, Mn deficiency caused DNA fragmentation, which might be partially associated with the up-regulation of the apoptosis signaling (caspase-3, caspase-8 and caspase-9) in the gills of fish. Furthermore, Mn deficiency-caused apoptosis might be partly related to the increases of oxidative damage that indicated by increases of lipid peroxidation and protein oxidation, and decreases of antioxidant enzyme activities [included Mn superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST)]. However, Mn deficiency only down-regulated MnSOD and GST mRNA levels, which might be partially related to the up-regulation of NF-E2-related factor-2 (Nrf2) inhibitor (Keap1), and only down-regulated the gene expression of claudin-b and claudin-15 to disrupt the TJ in the gills of fish. Excessive Mn led to negative effects on partial parameters studied in the gills of fish. The optimal levels of Mn based on protecting against ROS, MDA and PC in the gills of grass carp were 17.04, 16.86 and 21.20 mg/kg diet, respectively. Collectively, Mn deficiency or excess could cause inflammation, apoptosis, antioxidant system disruption and change tight junction protein (claudin-b and claudin-15) transcription abundances, which might be partially related to the NF-κB p65, caspase-(3,8,9) and Nrf2 signaling, in the gills of fish.

Antioxidant defenses in polar cod (Boreogadus saida) and responsiveness toward dietary crude oil exposure

Increasing anthropogenic activities in the Arctic pose the risk for accidental oil spills but our knowledge of ecotoxicological effects is still limited for Arctic organisms. This study aimed to provide insights in baseline levels of antioxidants in polar cod (Boreogadus saida) from different Arctic locations and investigate the susceptibility of this species to oxidative stress during a 32 day dietary crude oil exposure. Baseline levels of individual antioxidants and total oxyradical scavenging capacity (TOSC) varied among different Arctic fjords. Upon crude oil exposure, dose- and time-dependent polycyclic aromatic hydrocarbons (PAHs) biotransformation was evident through hepatic ethoxyresorufine-O-deethylase activity, glutathione S-transferase mRNA expression and biliary PAH metabolites. Changes in the activity of individual antioxidants and TOSC was, however, slight and transient upon exposure. In conclusion, biotransformation was clearly induced by the sub-lethal levels of crude oil in polar cod, while the antioxidant defense responded with high variability.

Interactive effect of ammonia and crowding stress on ion-regulation and expression of immune-related genes in juvenile turbot (Scophthalmus maximus)

The present study was to investigate the interactive effect of ammonia and crowding stress on ion-regulation and expression of immune-related genes in juvenile turbot (Scophthalmus maximus). The fish were exposed to four total ammonia nitrogen (0, 5, 20, 40 mg/L TAN) and two stocking density. After 96 h of exposure, blood, gill, and liver samples were collected to measure biochemical parameters and mRNA levels of immune-related genes. The results showed that co-exposure to high TAN (20 and 40 mg/L) and high density significantly increased plasma sodium (Na+), gill Na+/K+-ATPase activity and mRNA levels. Following individual and combined exposure to high TAN and high density, the heat shock protein 70 (HSP 70), HSP 90, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) genes expression were obviously higher than their control. Conversely, the lysozyme (LZM) and hepcidin mRNA levels were down-regulated in liver of fish exposed to high TAN alone and combination of high TAN-density. Moreover, glutathione S-transferase (GST) mRNA levels significantly increased in treatments with individual high TAN and high density, but decreased in high TAN-density co-exposed fish. Overall, ion homeostasis and immune status were adversely influenced in high exposed turbot under high density.