Expression Of Detoxification-related Genes Research Articles

Worldwide used bio-insecticides Cry1Ac toxin have no detrimental effects on E. balteatus but alter the symbiotic microbial communities.

Hoverflies, capable of abilities providing dual ecosystem services including pest control and pollination, are exposed to insecticidal proteins from transgenic plants via pollen and prey aphids. However, the effects of such exposures on hoverflies have never been adequately assessed. Here, we investigated impacts of the most widely used biotoxin Cry1Ac on a representative hoverfly species Episyrphus balteatus through food chain transmission and active toxin exposure. The results showed Cry1Ac can be transmitted into E. balteatus through feeding on Aphis gossypii reared by Bt insect-resistant cotton variety expressing the Cry1Ac toxin, but the biological parameters of E. balteatus including survival rate, growth, development, reproductive capacity, and detoxification-related gene expression, were not significantly affected. Furthermore, the exposure to high-dose Cry1Ac toxin (500 μg/mL) resulted in slight increase of 16.67 % in the activity of detoxification and antioxidant enzyme catalase in E. balteatus and inhibited the egg hatching, partially inducing stress responses. Notably, the exposure to Cry1Ac toxin disrupted the microbiota homeostasis in E. balteatus, and the relative abundances of three dominant symbiotic bacterial genera (Cosenzaea, Wolbachia, and Commensalibacter) in E. balteatus exhibited a 10 % ~ 40 % fluctuation under Cry1Ac toxin stress. Taken together, these results suggest Cry1Ac toxin is not lethal to E. balteatus, but it poses a potential threat to its endosymbiotic bacteria.

Toxicities of polystyrene microplastics (MPs) and hexabromocyclododecane (HBCD), alone or in combination, to the hepatopancreas of the whiteleg shrimp, Litopenaeus vannamei

The hepatopancreas is one of the largest organs playing crucial roles in metabolism and detoxification in crustacean invertebrates. Although toxicities have been increasingly documented for the two ubiquitous pollutants, hexabromocyclododecane (HBCD) and microplastics (MPs), in model animals, little is known about their impacts on the hepatopancreas of crustaceans. To fill this knowledge gap, the effects of MPs and HBCD, alone or in combination, on the hepatopancreas were evaluated in a commercially important crustacean species (the whiteleg shrimp) by histological observation as well as quantification of hepatic lesion-, metabolism-, and detoxification-related parameters. In addition, to reveal potential mechanisms underlying the hepatoxicity observed, the accumulation of HBCD in the shrimp and the status of oxidative stress were also investigated. Our results demonstrated that exposure of the whiteleg shrimp to MPs and HBCD for 4 weeks resulted in evident histological injury in the hepatopancreas and marked elevation in hepatic lesion markers (alanine aminotransferase and aspartate aminotransferase) in the hemolymph. Moreover, both metabolism (activity of phosphofructokinase, contents of lactic acid and adenosine triphosphate, and expression of metabolism-related genes) and detoxification (contents of cytochrome P450, UDP-glucuronosyltransferase, and glutathione, activity of glutathione S-transferase, and expression of detoxification-related genes) were found to be disrupted by the pollutants tested. In addition, exposure to MPs and HBCD also led to alterations in the contents and/or activities of antioxidant enzymes and resulted in oxidative damage to the hepatopancreas (indicated by marked elevation in malondialdehyde content). Furthermore, a significant amount of HBCD accumulated in shrimp treated with HBCD-containing seawater. The data also illustrated that HBCD-MP coexposure was more toxic than single exposure to these pollutants. These findings suggest that MPs and HBCD may exert hepatotoxic impacts on whiteleg shrimp by accumulating in vivo and inducing oxidative stress, which could pose a severe threat to the health of this important crustacean species.

Effects of tolfenpyrad exposure on development and response mechanism in the silkworm, Bombyx mori

Tolfenpyrad is a broad spectrum of insecticide that can effectively kill different types of pests, including Lepidoptera. However, due to improper use, the adverse effects of tolfenpyrad on beneficial or economic insects have not been well studied. In this study, we systematically investigated the toxic effect of sublethal tolfenpyrad on silkworms. Sublethal tolfenpyrad exposure can affect the body weight, developments days, cocooning rate, eclosion rate and pupation rate. To further study the response mechanism of silkworms to tolfenpyrad stimulation, we compared the different expression genes by transcriptome sequencing and verified them by qRT-PCR. We found that significant changes in the genes expression was involved in xenobiotics biodegradation and metabolism, immune system and digestive system after tolfenpyrad treatment. To further investigate the possible mechanisms by which intestinal microbia in the response to tolfenpyrad, we analysed the microbia changes in the midgut of silkworms by 16S rRNA gene sequencing. The results showed that the relative abundances of Enterobacter and Staphylococcus were increased whereas the Tyzzerella and Methylobacterium-Methylorubrum were decreased after tolfenpyrad stimulation. Taken together, these results indicated that low concentration of tolfenpyrad affect the growth and development of silkworms. Silkworms respond to the toxicity of tolfenpyrad by inducing immune and detoxification-related gene expression or altering microbial composition in the midgut.

Effects of glyphosate herbicide Roundup® on antioxidant enzymes activity and detoxification-related gene expression in honey bees (Apis mellifera)

Glyphosate (GLY) is a type of broad-spectrum herbicide and has been widely used for weed control. In the past few decades, GLY was generally considered to be low toxic or non-toxic to animals. However, recent studies demonstrated that GLY has adverse effects on honey bees. How honey bees’ internal antioxidant and detoxification system responds to GLY exposure needs to be investigated. In this study, we calculated the median lethal dose (LD50) of GLY in honey bees and evaluate the effect of GLY on detoxification-related gene expression and antioxidant enzyme activities of honey bees. Our results showed that the LD50 of GLY to honey bees at 24 and 48 h were 1773.06 and 537.67 µg active ingredient (a.i.)/bee, respectively, which suggested the low toxicity of GLY to honey bees. Moreover, we found significantly downregulated enzyme activity of superoxide dismutase (SOD), upregulated activities of glutathione S-transferase (GST) and acetylcholinesterase (AChE), as well as upregulated gene expression of CYP9Q1 in the head, and CYP9Q2 in the head and abdomen, which indicated the important roles of GST, AChE, and cytochrome P450 monooxygenase (CYPs/P450s) in protecting bees against GLY stress. Our study will help to clarify the role of antioxidative enzymes and detoxifying enzymes in terms of preventing or limiting GLY stress injury.

Effects of azadirachtin on detoxification-related gene expression in the fat bodies of the fall armyworm, Spodoptera frugiperda.

The fall armyworm, Spodoptera frugiperda, has become a worldwide pest and threatens world food production. A previous study indicated that azadirachtin, the most effective botanical insecticide for S. frugiperda, inhibits larval growth of the insect. The effect of azadirachtin on the tissues of the larvae, however, remains to be determined. In this study, the effects of azadirachtin on the structure of fat bodies were analyzed. Comparative transcriptomic analysis was conducted between controls and samples treated with 0.1μg/g azadirachtin for 7days to explore potential relevant mechanisms. The expression of 5356 genes was significantlyaffected after azadirachtin treatment, with 3020 up-regulated and 2336 down-regulated. Among them, 137 encode detoxification enzymes, including 53 P450s, 20 GSTs, 27 CarEs, 16 UGTs, and 12 ABC transporters. Our results indicated that azadirachtin could destroy fat body structure and change the mRNA levels of detoxification-related genes. The up-regulated genes encoding detoxification enzymes might be related to detoxifying azadirachtin. Our results elucidate a preliminary mechanism of azadirachtin detoxification in the fat bodies of S. frugiperda larvae.

Adverse effects in Daphnia magna exposed to e-waste leachate: Assessment based on life trait changes and responses of detoxification-related genes

The sharp increase in e-waste derived from great consumption of electronic products has become a potential global environmental challenge. Limited information is available about the potential detrimental impact of e-waste on aquatic organisms. The present study investigated the expression of detoxification-related genes and life-history parameter changes in Daphnia magna exposed to e-waste leachate, simultaneously integrating with the chemical analysis of typical pollutants from e-waste leachate. The study aims at assessing impacts of e-waste to aquatic invertebrates and providing insights into its toxic mechanisms. The results showed high concentration of heavy metals like Cu (1657.14 ± 259.3 μg g−1, DW) and persistent organic compounds like polybrominated diphenyl ethers (7831.32 ± 1273.86 ng g−1, DW) in stream sediments near e-waste dismantling areas. Chronic exposure to these pollutants can affect the growth and reproduction of D. magna, resulting in significant development retardation, decreased total egg production per female, and even smaller body size. Expression of some detoxification and reproduction-related genes including DappuHR96, CYP360a, P-gp, EcR, CYP314 and Vtg exhibited different response patterns depending on the e-waste leachate concentration. E-waste leachate may affect the expression of detoxification-related and growth and reproduction-related genes and disrupt the growth and reproduction processes of D. magna.

Effect of environmental heavy metals on the expression of detoxification-related genes in honey bee Apis mellifera

Air pollutants and agricultural pesticides can be environmental stressors to pollinators. In this study, to investigate the expression of detoxification-related genes and heavy metal concentrations in honey bees and honey possibly exposed to environmental stresses, we collected samples from apiaries located in mountainous, agricultural, and urban areas. Compared with the mountainous and agricultural areas, the mercury and lead concentrations were highest in honey and bees collected from urban areas. In addition, the expression levels of CYP9Q1, CYP9Q2, CYP9Q3, and genes encoding catalase and superoxide dismutase were markedly higher in urban bees than those from agricultural and mountainous areas, discreetly indicating that the notable induction of the detoxification metabolism in urban bees might be because of heavy metal pollutant exposure. Our study suggests that honey bees actively respond to environmental stressors, such as heavy metals derived from urban areas.

Detoxification-related gene expression accompanies anhydrobiosis in the foliar nematode (Aphelenchoides fragariae)

The foliar nematode (Aphelenchoides fragariae) is a quarantined pest that infects a broad range of herbaceous and woody plants. Previous work has demonstrated its remarkable ability to survive rapid and extreme desiccation, although the specific molecular mechanisms underlying its anhydrobiotic response have not been characterized. The authors used RNA sequencing and de novo transcriptome assembly to compare patterns of gene expression between hydrated and 24-hr desiccated nematodes. In total, 2,083 and 953 genes were significantly up- and downregulated, respectively, in desiccated nematodes. Of the 100 annotated genes with the largest positive fold-changes, more than one third encoded putative detoxification-related proteins. Genes encoding enzymes of Phase I and Phase II detoxification systems were among the most strongly upregulated in the transcriptome, including 35 cytochrome p450s, 23 short chain dehydrogenase/reductases, 5 glutathione-S-transferases, and 22 UDP-glucuronosyltransferases. Genes encoding heat shock proteins, unfolded protein response enzymes, and intrinsically disordered proteins were also upregulated. Anhydrobiosis in A. fragariae appears to involve both strategies to minimize protein misfolding and aggregation, and wholesale induction of the cellular detoxification machinery. These processes may be controlled in part through the activity of forkhead transcription factors similar to Caenorhabditis elegans’ daf-16, a number of which were differentially expressed under desiccation.

The host generalist phytopathogenic fungus Sclerotinia sclerotiorum differentially expresses multiple metabolic enzymes on two different plant hosts

Sclerotinia sclerotiorum is a necrotrophic fungal pathogen that infects upwards of 400 plant species, including several economically important crops. The molecular processes that underpin broad host range necrotrophy are not fully understood. This study used RNA sequencing to assess whether S. sclerotiorum genes are differentially expressed in response to infection of the two different host crops canola (Brassica napus) and lupin (Lupinus angustifolius). A total of 10,864 of the 11,130 genes in the S. sclerotiorum genome were expressed. Of these, 628 were upregulated in planta relative to in vitro on at least one host, suggesting involvement in the broader infection process. Among these genes were predicted carbohydrate-active enzymes (CAZYmes) and secondary metabolites. A considerably smaller group of 53 genes were differentially expressed between the two plant hosts. Of these host-specific genes, only six were either CAZymes, secondary metabolites or putative effectors. The remaining genes represented a diverse range of functional categories, including several associated with the metabolism and efflux of xenobiotic compounds, such as cytochrome P450s, metal-beta-lactamases, tannases and major facilitator superfamily transporters. These results suggest that S. sclerotiorum may regulate the expression of detoxification-related genes in response to phytotoxins produced by the different host species. To date, this is the first comparative whole transcriptome analysis of S. sclerotiorum during infection of different hosts.

Interactive effects of diclofenac and copper on bioconcentration and multiple biomarkers in crucian carp (Carassius auratus).

Diclofenac (DCF), a non-steroidal anti-inflammatory drug, is widespread in aquatic environments and coexists with heavy metals to form combined pollution. However, the interactive effects of DCF and heavy metals on aquatic organisms remain unknown. This study aimed to investigate the interactive effects of DCF and copper (Cu) on the bioconcentration, oxidative stress status and detoxification-related gene expression in crucian carp (Carassius auratus). Fish were exposed to Cu (100 μg L-1) and DCF (1, 10, 100 and 1000 μg L-1) alone or in combination for 7 days. Results obtained showed that the treatment of Cu combined with high levels of DCF (100 and 1000 μg L-1) significantly decreased tissue concentrations of DCF and Cu compared to the correspondingly individual exposure. Concerning oxidative stress status, as reflected by the activities of antioxidant enzymes and malondialdehyde content, low exposure concentrations of DCF (1 and 10 μg L-1) seemed to mitigate the oxidative stress induced by Cu, whereas the co-exposure of Cu with the highest level of DCF (1000 μg L-1) led to stronger oxidative damage in fish liver than Cu exposure alone. With regarding to detoxification-related genes, in most cases, the expressions of cyp 1a, cyp 3a, gstα, gstπ, pxr and P-gp in crucian carp were significantly altered upon exposure to the compounds in combination compared to exposure to the compounds individually. Collectively, these findings indicate the capacity of each of these pollutants to alter bioconcentration potential, pro-oxidative effects and detoxification-related gene responses of the other when both co-occur at specific concentrations.

Dietary curcumin enhances intestinal antioxidant capacity in ducklings via altering gene expression of antioxidant and key detoxification enzymes.

The study investigated the effects of dietary curcumin supplementation on tissue distribution of curcumin and its metabolites, intestinal antioxidant capacity, and expression of detoxification-related genes in ducks. A total of 720 one-day-old male Cherry Valley Pekin ducklings (initial BW 58.6 ± 0.1g) were randomly assigned to 4 dietary groups each with 6 replicates of 30 ducks using a single factorial arrangement design. Ducks in the control group were fed a basal diet and the remainder were fed the basal diet supplemented with 200, 400, or 800mg/kg curcumin. The experiment lasted for 21 D. Curcumin was present at 13.12 to 16.18mg/g in the cecal digesta, 75.50 to 575.40μg/g in jejunal mucosa, 35.10 to 73.65μg/g in liver, and 7.02 to 7.88μg/mL in plasma. The jejunal and hepatic contents of curcumin increased significantly (P < 0.05) in response to supplementation with 400 and 800mg/kg of curcumin respectively, compared with 200mg curcumin/kg group. There was a linear (P < 0.001) effect of dietary curcumin on relative abundance of SOD1, GPX1, CAT, HO-1, and Nrf2 transcripts, and a quadratic (P < 0.001) increase in the activities of GSH-Px and T-AOC in jejunal mucosa. The expression of CYP1A4, CYP2D17 increased and CYP1B1, CYP2A6 decreased linearly (P < 0.001) with dietary curcumin concentrations. In addition, dietary curcumin increased gene expression of GST, MRP6, and ABCB1 in jejunal mucosa. In conclusion, dietary supplementation with 200 to 800mg/kg curcumin enhanced the accumulation of curcumin and its metabolites in jejunum as well as increasing the antioxidant capacity and detoxification potential, which play major roles in the protection of duck intestines against damage.

Simvastatin affect the expression of detoxification-related genes and enzymes in Daphnia magna and alter its life history parameters

Simvastatin (SV), as an hypocholesterolaemic drug, has been detected in various aquatic environment. However, limited information is available on the effects of SV on freshwater invertebrates. In the present study, we investigated the toxic effects of SV on Daphnia. magna (D. magna) through measuring the physiological changes (e.g., survival, growth rate, and reproduction) in a 21-d chronic toxicity test We also determined the expression of seven detoxification and reproduction-related genes (i.e. HR96, P-gp, CYP360A8, GST, CYP314, EcR and Vtg) and several enzymes (i.e. APND, ERND, GST and CAT) in a acute test (24 h). Results showed that high concentration (e.g. 50 μg L−1) of SV for short time exposure (e.g. 24 h) significantly induced the expression of HR96 and P-gp (e.g. up to 2.5 folds)and enzymes (e.g. increasing 4.0 folds for ERND and GST activity) in D. magna.. The long-term chronic exposure (21 days) may cause the changes of life history parameters such as decreasing total egg production number per individual and intrinsic growth rates etc. SV may act as a potential endocrine disruptor to D. magna and the reproduction parameters were more sensitive endpoints than the survival and growth for evaluating SV exposure.

Effects of chlorantraniliprole exposure on detoxification enzyme activities and detoxification-related gene expression in the fat body of the silkworm, Bombyx mori

Chlorantraniliprole (CAP) can induce excessive calcium release from muscle of insects, causing muscle paralysis until death, and its residues in farmland can cause poisoning in Bombyx mori (B. mori), resulting in the failure of cocooning. No reports have investigated the effects of CAP exposure on detoxification enzyme activities and detoxification-related gene expression in B. mori. In the present study, we treated mulberry leaves with CAP by the leaf-dipping method, and then B. mori larvae were continuously fed with the polluted mulberry leaves. Moreover, the detoxification enzyme activities and the expressions of detoxification-related genes in the fat body of B. mori were examined. The results showed that at 24 h after CAP exposure, the activities of P450 and GST enzymes were all significantly increased, with P450 enzymes responding fastest. CarE enzyme activity was up-regulated in 24 h, and then it was decreased compared with the control group. Furthermore, the expressions of the key genes in the PI3K/Akt/CncC signaling pathway (PI3K, PDK, Akt, CncC and Keap1) at the mRNA were significantly increased. Western blotting analysis revealed that Akt was inhibited at the protein level, resulting in decreased expression of Keap1 and increased expression of CncC. These results indicated that the PI3K/Akt/CncC signaling pathway in the fat body of B. mori responded to CAP exposure and regulated the expressions of downstream detoxification enzymes, thus enhancing the detoxifying capability of B. mori.

RNA-Seq transcriptome analysis of Amaranthus palmeri with differential tolerance to glufosinate herbicide.

Amaranthus palmeri (Amaranthaceae) is a noxious weed in several agroecosystems and in some cases seriously threatens the sustainability of crop production in North America. Glyphosate-resistant Amaranthus species are widespread, prompting the use of alternatives to glyphosate such as glufosinate, in conjunction with glufosinate-resistant crop cultivars, to help control glyphosate-resistant weeds. An experiment was conducted to analyze the transcriptome of A. palmeri plants that survived exposure to 0.55 kg ha-1 glufosinate. Since there was no record of glufosinate use at the collection site, survival of plants within the population are likely due to genetic expression that pre-dates selection; in the formal parlance of weed science this is described as natural tolerance. Leaf tissues from glufosinate-treated and non-treated seedlings were harvested 24 h after treatment (HAT) for RNA-Seq analysis. Global gene expression was measured using Illumina DNA sequence reads from non-treated and treated surviving (presumably tolerant, T) and susceptible (S) plants. The same plants were used to determine the mechanisms conferring differential tolerance to glufosinate. The S plants accumulated twice as much ammonia as did the T plants, 24 HAT. The relative copy number of the glufosinate target gene GS2 did not differ between T and S plants, with 1 to 3 GS2 copies in both biotypes. A reference cDNA transcriptome consisting of 72,780 contigs was assembled, with 65,282 sequences putatively annotated. Sequences of GS2 from the transcriptome assembly did not have polymorphisms unique to the tolerant plants. Five hundred sixty-seven genes were differentially expressed between treated T and S plants. Of the upregulated genes in treated T plants, 210 were more highly induced than were the upregulated genes in the treated S plants. Glufosinate-tolerant plants had greater induction of ABC transporter, glutathione S-transferase (GST), NAC transcription factor, nitronate monooxygenase (NMO), chitin elicitor receptor kinase (CERK1), heat shock protein 83, ethylene transcription factor, heat stress transcription factor, NADH-ubiquinone oxidoreductase, ABA 8’-hydroxylase, and cytochrome P450 genes (CYP72A, CYP94A1). Seven candidate genes were selected for validation using quantitative real time-PCR. While GST was upregulated in treated tolerant plants in at least one population, CYP72A219 was consistently highly expressed in all treated tolerant biotypes. These genes are candidates for contributing tolerance to glufosinate. Taken together, these results show that differential induction of stress-protection genes in a population can enable some individuals to survive herbicide application. Elevated expression of detoxification-related genes can get fixed in a population with sustained selection pressure, leading to evolution of resistance. Alternatively, sustained selection pressure could select for mutation(s) in the GS2 gene with the same consequence.

Hidden genetic variation shapes the structure of functional elements in Drosophila

Mutations that add, subtract, rearrange, or otherwise refashion genome structure often affect phenotypes, although the fragmented nature of most contemporary assemblies obscures them. To discover such mutations, we assembled the first new reference-quality genome of Drosophila melanogaster since its initial sequencing. By comparing this new genome to the existing D. melanogaster assembly, we created a structural variant map of unprecedented resolution and identified extensive genetic variation that has remained hidden until now. Many of these variants constitute candidates underlying phenotypic variation, including tandem duplications and a transposable element insertion that amplifies the expression of detoxification-related genes associated with nicotine resistance. The abundance of important genetic variation that still evades discovery highlights how crucial high-quality reference genomes are to deciphering phenotypes.

Low levels of Cd induce persisting epigenetic modifications and acclimation mechanisms in the earthworm Lumbricus terrestris.

Toxic effects of cadmium (Cd), a common soil pollutant, are still not very well understood, particularly in regard to its epigenetic impact. Therefore, the aim of this study was to assess DNA methylation changes and their persistence in the earthworm Lumbricus terrestris upon chronic low dose Cd exposure using methylation sensitive amplification polymorphism (MSAP). Moreover, the biomarker response and fitness of the earthworms, as well as the expression of detoxification-related genes (metallothionein (MT) and phytochelatin synthase (PCS)) was evaluated. Low levels of Cd caused an increase in genome-wide DNA methylation, which remained partly modified, even after several months of recovery in unpolluted soil. Increased cellular stress seemed to decrease after two weeks of exposure whereas fitness parameters remained unaffected by Cd, probably as a result from the activation of detoxification mechanisms like the expression of MTs. Interestingly, even though the level of Cd exposure was very low, MT expression levels indicate the development of acclimation mechanisms. Taken together, this study demonstrates that acclimation, as well as epigenetic modifications can occur already in moderately polluted environments. In addition, these effects can have long-lasting impacts on key species of soil invertebrates and might persist long after the actual heavy metal challenge has passed.

Toxic effects of diclofenac on life history parameters and the expression of detoxification-related genes in Daphnia magna

Diclofenac (DCF), as a widely used drug, has been detected in various environmental media such as municipal wastewater effluent. However, there is little information on the effects of DCF on freshwater invertebrates potentially exposing to its residues in surface water. In the present study, we investigated the toxic effects of DCF on the physiological parameters (e.g., survival, growth rate, and reproduction) of a crustacean, Daphnia magna, via a 21-d chronic toxicity test, and we also evaluated the effects of DCF on the expression of the genes related to the detoxification metabolism, growth, development and reproduction (e.g., HR96, P-gp, CYP360A8, CYP314, GST, EcR and Vtg) in acute exposure (up to 96h) with RT-PCR. Significant toxic effects of DCF to D. magna were observed at 50μgL−1, the expression of these selected genes was inhibited with 24h of exposure, and induced after 48h to some extents. The expression of Vtg was induced at high concentrations of DCF (500μgL−1 and 5000μgL−1) after 24h and 48h of exposure, but also significantly induced at low concentration (50μgL−1) after 96h of exposure. Dose- and time-dependent relationships were observed for gene expression of the seven selected genes. In the 21-d chronic toxicity test, the days to the first brood and the days to the first egg production were both significantly delayed at 50μgL−1. However, there were no significant differences observed among the molting frequency, number of eggs produced in the first brood, total number of eggs per individual, total number of broods per individual, body length and intrinsic growth rate. Our results suggested that the reproduction parameters are more sensitive endpoints than the survival and growth for evaluating the toxicity of DCF to aquatic invertebrates.

Broccoli sprout extract induces detoxification-related gene expression and attenuates acute liver injury.

To investigate the effects of broccoli sprout extract (BSEx) on liver gene expression and acute liver injury in the rat. First, the effects of BSEx on liver gene expression were examined. Male rats were divided into two groups. The Control group was fed the AIN-76 diet, and the BSEx group was fed the AIN-76 diet containing BSEx. After a 10-d feeding period, rats were sacrificed and their livers were used for DNA microarray and real-time reverse transcription-polymerase chain reaction (RT-PCR) analyses. Next, the effects of BSEx on acute liver injury were examined. In experiments using acute liver injury models, 1000 mg/kg acetaminophen (APAP) or 350 mg/kg D-galactosamine (D-GalN) was used to induce injury. These male rats were divided into four groups: Control, BSEx, Inducer (APAP or D-GalN), and Inducer+BSEx. The feeding regimens were identical for the two analyses. Twenty-four hours following APAP administration via p.o. or D-GalN administration via i.p., rats were sacrificed to determine serum aspartate transaminase (AST) and alanine transaminase (ALT) levels, hepatic glutathione (GSH) and thiobarbituric acid-reactive substances accumulation and glutathione-S-transferase (GST) activity. Microarray and real-time RT-PCR analyses revealed that BSEx upregulated the expression of genes related to detoxification and glutathione synthesis in normal rat liver. The levels of AST (70.91 ± 15.74 IU/mL vs 5614.41 ± 1997.83 IU/mL, P < 0.05) and ALT (11.78 ± 2.08 IU/mL vs 1297.71 ± 447.33 IU/mL, P < 0.05) were significantly suppressed in the APAP + BSEx group compared with the APAP group. The level of GSH (2.61 ± 0.75 nmol/g tissue vs 1.66 ± 0.59 nmol/g tissue, P < 0.05) and liver GST activity (93.19 ± 16.55 U/g tissue vs 51.90 ± 16.85 U/g tissue, P < 0.05) were significantly increased in the APAP + BSEx group compared with the APAP group. AST (4820.05 ± 3094.93 IU/mL vs 12465.63 ± 3223.97 IU/mL, P < 0.05) and ALT (1808.95 ± 1014.04 IU/mL vs 3936.46 ± 777.52 IU/mL, P < 0.05) levels were significantly suppressed in the D-GalN + BSEx group compared with the D-GalN group, but the levels of AST and ALT in the D-GalN + BSEx group were higher than those in the APAP + BSEx group. The level of GST activity was significantly increased in the D-GalN + BSEx group compared with the D-GalN group (98.04 ± 15.75 U/g tissue vs 53.15 ± 8.14 U/g tissue, P < 0.05). We demonstrated that BSEx protected the liver from various types of xenobiotic substances through induction of detoxification enzymes and glutathione synthesis.

Effects of steroid hormones on reproduction- and detoxification-related gene expression in adult male mosquitofish, Gambusia affinis

The molecular mechanisms that mediate fish reproduction and detoxification in response to steroid hormones were studied by using adult male western mosquitofish (Gambusia affinis) as sentinel species. The expression patterns of three vitellogenins (VtgA, VtgB and VtgC), two estrogen receptors (ERα and ERβ), two androgen receptors (ARα and ARβ), metallothionein (MT) and cytochrome P450 1A (CYP1A) in the liver and testis of adult male mosquitofish were assessed through exposure treatments with progesterone (P), testosterone (T) and 17β-estradiol (E2), alone and in combination for eight days. The results showed that expression patterns of Vtg subtype, ER subtype, AR subtype, MT and CYP1A genes in male mosquitofish varied according to tissue and specific hormone stress. Vtg subtype mRNA expression was induced in the liver in E2-added treatments, and an up-regulation of ERα mRNA expression was also observed. In addition, hormone treatments increased three Vtg subtype mRNA expression levels in the testis, at least to some extent. All hormone treatments significantly inhibited ERα, ERβ and ARβ mRNA expression in the testis. Some of hormone treatments could affect MT and CYP1A gene expression in mosquitofish. In general, multiple hormone treatments showed different effects on target gene expression compared with corresponding hormone alone. The results from the present study provided valuable information on the toxicological effects of steroid hormones in mosquitofish.

Effects of dietary tert-butylhydroquinone on domoic acid metabolism and transcription of detoxification-related liver genes in red sea bream Pagrus major

Domoic acid (DA) is a neuroexcitatory amino acid that is produced by Pseudo-nitzschia during harmful algal blooms (HAB). Accumulation of DA can be transferred through food chain and cause neuronal damage in marine animal and in human. Like other algal toxins, DA was suggested to increase the oxidative stress and increase the detoxification-related gene expression in fish. The widely used food antioxidant, tert-butylhydroquinone (tBHQ), was known to induce a wide range of antioxidative potentials such as elevation of the glutathione levels and glutathione S-transferases (GSTs), via the activation of antioxidant response elements (AREs). In this study, the influences of dietary tBHQ on domoic acid (DA) metabolism and detoxification-related gene transcription were investigated both in vivo and in vitro. Oral administration of tBHQ resulted in significant decreases of DA accumulation of liver tissues in which red sea bream were fed with a single dose of 10 mg DA and 100 mg tBHQ per kg body weight per fish. Real-time PCR further revealed that the mRNA levels of AHR/ARNT/CYP1A1/GSTA1/GSTR were up-regulated in the above liver tissues at 72 h post tBHQ treatment. In consistence, tBHQ exposure also resulted in increased mRNA transcription of GSTA1, GSTA2 and GSTR in cultured red sea bream hepatocytes. Collectively, our findings in this research suggested that the dietary intake of tBHQ accelerated DA metabolism in fish, through mechanisms involving altered transcription of detoxification-related liver genes.