RESUMO
The mechanism by which an organism can adapt to subtle environmental changes is predicated on modifications to biochemical processes within the cellular metabolism in response to such changes. Changes in these processes have the potential to induce alterations in cellular structures and tissue organization, as well as establish a causal link between fluctuations in these parameters and stressors exposure. This investigation's main goal and innovation is to evaluate the environmental stress indicators in the aquatic ecosystem of Lake Qarun. Pesticide residues in freshwater fish should be the primary focus of evaluation of environmental stressor concentrations, since they serve as bioindicators at different times and places on a spatiotemporal scale. A thorough analysis of suggestive biochemical biomarker reactions should also be conducted. The effects of environmental stressors, specifically pesticide contamination in Qarun Lake, have been observed and investigated in relation to two fish species: Solea aejabtiaca and Oreochronis niloticus. The results of a hazard assessment conducted at six sampling sites using spatio-temporal data revealed elevated mean values for the pesticides, persistent organic pollutants (POPs), organochlorines, organophosphates, and pyrethroids that were detected. A multi biomarker approach facilitates a more comprehensive understanding of stress responses induced by exposure to pollutants. As a result, the activities of the biochemical biomarkers CYP-450, GST, GSH, and LDH in the blood and liver of fish samples were found to be notably elevated. The suitability of the identified variables for biomonitoring of aquatic pollution was validated, and the data unveiled variations in sensitivity among species, implying that Nile tilapia could potentially function as a bioindicator with high sensitivity. The findings were correlated with the concentrations of detrimental organochlorines, organophosphorus, and pyrethroids in the muscles and gills. The data indicates that pollutants linked to agricultural wastes, runoff, and municipal effluent may be discharged into the lake ecosystem. Consequently, to safeguard the environment, it is essential to enforce and implement policies, acts, and regulations that already exist. Assessing the effects of additional environmental stressors on aquatic ecosystems is another way in which biomarker screening with an integrative approach improves our comprehension of how toxicants impact various levels of biological organization and is particularly useful in realistic environmental exposure scenarios.
Assuntos
Biomarcadores , Monitoramento Ambiental , Poluentes Químicos da Água , Animais , Biomarcadores/metabolismo , Poluentes Químicos da Água/toxicidade , Monitoramento Ambiental/métodos , Lagos , Peixes/metabolismo , Estresse Fisiológico , Praguicidas/toxicidade , Fígado/metabolismo , Fígado/efeitos dos fármacos , Água Doce , Ciclídeos/metabolismo , Glutationa Transferase/metabolismo , Resíduos de Praguicidas/toxicidade , Resíduos de Praguicidas/análiseRESUMO
The black cutworm, Agrotis ipsilon (Lepidoptera: Noctuidae), is an important agricultural pest. Phoxim is an organophosphate insecticide that has been widely used to control A. ipsilon. The extensive application of phoxim has resulted in a reduction in phoxim susceptibility in A. ipsilon. However, the molecular mechanisms underlying phoxim tolerance in A. ipsilon remain unclear. In this work, we report the involvement of AiGSTz1, a zeta class glutathione S-transferase, in phoxim tolerance in A. ipsilon. Exposure to a sublethal concentration (LC50) of phoxim dramatically upregulated the transcription level of the AiGSTz1 gene in A. ipsilon larvae, and this upregulation might be caused by phoxim-induced oxidative stress. The recombinant AiGSTz1 protein expressed in Escherichia coli was able to metabolize phoxim. Furthermore, AiGSTz1 displayed antioxidant activity to protect against oxidative stress. Knockdown of AiGSTz1 by RNA interference significantly increased the mortality rate of A. ipsilon larvae in response to phoxim. In addition, the transcription factor AiCncC can bind to the cap 'n' collar isoform C: muscle aponeurosis fibromatosis (CncC:Maf) binding site in the putative promoter of the AiGSTz1 gene. Silencing of AiCncC resulted in a dramatic downregulation of AiGSTz1. These results indicated that AiGSTz1 is involved in phoxim tolerance and is potentially regulated by AiCncC. These findings provide valuable insights into the defense mechanisms used by A. ipsilon against phoxim.
Assuntos
Glutationa Transferase , Proteínas de Insetos , Inseticidas , Mariposas , Compostos Organotiofosforados , Fatores de Transcrição , Animais , Glutationa Transferase/metabolismo , Glutationa Transferase/genética , Compostos Organotiofosforados/farmacologia , Compostos Organotiofosforados/toxicidade , Inseticidas/farmacologia , Inseticidas/toxicidade , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Mariposas/efeitos dos fármacos , Mariposas/genética , Proteínas de Insetos/metabolismo , Proteínas de Insetos/genética , Larva/efeitos dos fármacos , Resistência a Inseticidas/genética , Estresse Oxidativo/efeitos dos fármacosRESUMO
Stresses caused by ionizing radiation can also damage tissues and organs through the circulatory system. In this study, we aimed to determine the radioprotective effect of propolis, a natural and powerful antioxidant product, against oxidative liver damage caused by cranial irradiation. Thirty-two male albino Sprague-Dawley rats, divided into four groups, were designed as sham group, irradiation (IR) group, propolis plus IR, control group of propolis. Biochemical parameters were measured in liver tissue of rats. While Total enzymatic superoxide scavenging activity (TSSA) and non-enzymatic superoxide scavenging activity (NSSA), glutathione peroxidase (GSH-Px) activities of all groups were statistically significantly higher than rats receiving only-irradiation, Glutathione-S-transferase (GST) activity in the IR group was significantly lower than in the sham control group and IR + propolis group. Superoxide dismutase (SOD) activity in the IR group was found to be significantly higher than both the sham control group and the propolis control group, but lower than the IR + propolis group. Malondialdehyde level and xanthine oxidase activity were higher in the IR group than in the other groups. Compared to the sham control group, in the group treated with propolis, a significant elevation in antioxidant parameters, specifically TSSA, NSSA, SOD, and GST activities, was noted, with corresponding increases of 32.3%, 23.2%, 47.6%, and 22.6%, respectively. Our findings show that propolis can be a radioprotective agent against ionized radiation damage by increasing antioxidant activity and reducing oxidant stress in liver tissue.
Assuntos
Antioxidantes , Fígado , Estresse Oxidativo , Própole , Protetores contra Radiação , Ratos Sprague-Dawley , Animais , Própole/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Fígado/metabolismo , Fígado/efeitos dos fármacos , Fígado/efeitos da radiação , Masculino , Ratos , Protetores contra Radiação/farmacologia , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Superóxido Dismutase/metabolismo , Glutationa Peroxidase/metabolismo , Malondialdeído/metabolismo , Glutationa Transferase/metabolismo , Xantina Oxidase/metabolismoRESUMO
The effectiveness of bensulfuron-methyl in controlling Schoenoplectiella juncoides (Roxb.) Lye has significantly decreased in rice fields in China. Hence, a bensulfuron-methyl-resistant S. juncoides population (W15) was collected from Dandong City, Liaoning Province, China, to investigate the underlying resistance mechanisms. Whole-plant dose-response experiments and ALS activity assay confirmed that W15 has evolved high-level resistance to bensulfuron-methyl compared with the susceptible S. juncoides population (W4). Molecular analysis revealed a Pro-197-Ser mutation in ALS1, while there was no significant difference in the relative ALS gene expression between W15 and W4. LC-MS/MS analysis showed W15 metabolized bensulfuron-methyl more rapidly than W4. Furthermore, bensulfuron-methyl resistance in W15 was significantly alleviated by malathion and 4-chloro-7-nitrobenzoxadiazole (NBD-Cl). Glutathione S-transferase activity was higher in W15 than in W4. Meanwhile, W15 displayed cross-resistance to halosulfuron-methyl and multi-resistance to MCPA-Na. In summary, these findings demonstrated for the first time that both target- and non-target-site resistance are relevant in the resistance of S. juncoides to bensulfuron-methyl.
Assuntos
Compostos de Sulfonilureia , Compostos de Sulfonilureia/farmacologia , Resistência a Herbicidas/genética , Herbicidas/farmacologia , Mutação , Glutationa Transferase/metabolismo , Glutationa Transferase/genéticaRESUMO
Eleusine indica is one of the most troublesome weeds in farmland worldwide, especially in Citrus Orchard of China. Glufosinate, as an efficient non-selective broad-spectrum herbicide, has been widely utilized for the control of E. indica in Citrus Orchard. The E. indica resistant population (R) was collected from a Citrus Orchard in Yichang City in Hubei province, China. Bioassay experiments showed that the R plants exhibited 3-fold resistance to glufosinate compared with the E. indica susceptible population (S). No known glutamine synthetase (GS) gene mutation associated with glufosinate resistance was found in R plants. And there was also no significant difference in GS activity between R and S plants. Those results indicated that the resistance to glufosinate in R did not involve target-site resistance. However, glutathione S-transferase (GST) inhibitor 4-chloro-7-nitrobenzoxadiazole (NBD-Cl) plus glufosinate gave a better control of R plants compared with glufosinate treatment alone. Moreover, both before and after glufosinate treatment, the GST activity in R plants was significantly higher than that in S plants. By RNA-seq, the expression of GSTU6 and GST4 up-regulated in R plants relative to S plants with or without glufosinate treatment. They were also significantly up-regulated expression in E. indica field resistant populations compared with S population. In summary, the study elucidated that R plants developed metabolic resistance to glufosinate involving GST. And GSTU6 and GST4 genes may play an important role in this glufosinate metabolic resistance. The research results provide a theoretical basis for a deeper understanding of resistance mechanism to glufosinate in E. indica.
Assuntos
Aminobutiratos , Eleusine , Resistência a Herbicidas , Herbicidas , Aminobutiratos/farmacologia , Herbicidas/farmacologia , Resistência a Herbicidas/genética , Eleusine/genética , Eleusine/metabolismo , Eleusine/efeitos dos fármacos , Glutationa Transferase/metabolismo , Glutationa Transferase/genética , Glutamato-Amônia Ligase/metabolismo , Glutamato-Amônia Ligase/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Herbicides are the main class of pesticides applied in crops and are capable of polluting the surrounding freshwater system; thus, understanding their impact on non-target species, whose mechanism of action is not described, helps to elucidate the real risks of these pollutants to the environment. 2,4-dichlorophenoxyacetic acid (2,4-D) is frequently detected in water and, due to its persistence, poses a risk to wildlife. In this way, the present work aimed to describe the implication of exposure to concentrations of 2,4-D already reported in aquatic environments in several physiological mechanisms of C. riparius at molecular and biochemical levels. To achieve this, bioassays were conducted with fourth instar larvae exposed to three concentrations of 2,4-D (0.1, 1.0, and 7.5 µg L-1). Larvae were collected after 24 and 96 h of exposure, and the expression of 42 genes, related to six subcellular mechanisms, was assessed by Real-Time PCR (RT-PCR). Besides, the activity of the enzymes catalase (CAT), glutathione S-transferase (GST), and acetylcholinesterase (AChE) was determined. The main metabolic route altered after exposure to 2,4-D was the endocrine system (mainly related to 20-hydroxyecdysone and juvenile hormone), confirming its endocrine disruptor potential. Four of the eleven stress response genes studied were down-regulated, and later exposure modulated DNA-repair genes suggesting genotoxic capacity. Moreover, only one gene from each detoxification phase was modulated at short exposure to 1.0 µg L-1. The molecular responses were not dose-dependent, and some early responses were not preserved after 96 h, indicating a transient response to the herbicide. Exposure to 2,4-D did not alter the activity of CAT, GST, and AChE enzymes. The responses described in this study reveal new mechanistic pathways of toxicity for 2,4-D in non-target organisms and highlight potential ecological consequences for chironomids in aquatic systems at the edges of agricultural fields.
Assuntos
Ácido 2,4-Diclorofenoxiacético , Chironomidae , Glutationa Transferase , Herbicidas , Ácido 2,4-Diclorofenoxiacético/toxicidade , Animais , Chironomidae/efeitos dos fármacos , Chironomidae/genética , Herbicidas/toxicidade , Glutationa Transferase/metabolismo , Glutationa Transferase/genética , Acetilcolinesterase/metabolismo , Acetilcolinesterase/genética , Larva/efeitos dos fármacos , Larva/genética , Larva/metabolismo , Poluentes Químicos da Água/toxicidade , Catalase/metabolismo , Catalase/genética , Expressão Gênica/efeitos dos fármacosRESUMO
The invasive golden apple snail Pomacea canaliculata is one of the devastating threats to aquatic ecosystems and wetland agriculture worldwide. Macrolides from microbes display various advantages over other compounds in controlling snails. However, emergence of antibiotic-resistant phenotypes against certain macrolides in the field appeals for exploring more effectively molluscicidal macrolides. Here, two borrelidins, borrelidin BN1 and BN2, from the extract of a Streptomyces strain fermentation were evaluated for molluscicidal potential against P. canaliculata using both immersion and contact bioassay methods. Borrelidin BN1 (borrelidin A) presented a significant molluscicidal activity comparable to the chemical pesticide metaldehyde, and had a much lower median lethal concentration value (LC50, 522.984 µg·ml-1) than avermectin B1 at 72 h of contact-killing treatment. Snail growth was inhibited by borrelidin BN1 more than by metaldehyde at sublethal concentrations, consistent with responses of key biochemical parameters. Exposure to borrelidin BN1 decreased the activity of acetylcholinesterase (AChE), glutathione S-transferase (GST), aspartate aminotransferase (AST), alanine aminotransferase (ALT) as well as the levels of energy reserves and sex steroids in snail tissues, while increased the activity of superoxide dismutase (SOD), catalase (CAT), lactate dehydrogenase (LDH) and the level of lipid peroxidation (LPO). Further application assay confirmed that borrelidin BN1 protected crop plant Zizania latifolia from P. canaliculata damage via suppressing snail population density. These findings suggest great potential of borrelidin BN1 as a molluscicide. Additionally, its higher activity than the stereoisomeric borrelidin BN2 (borrelidin F) implied better molluscicidal borrelidins could be acquired through structural optimization.
Assuntos
Moluscocidas , Caramujos , Animais , Moluscocidas/farmacologia , Caramujos/efeitos dos fármacos , Acetilcolinesterase/metabolismo , Compostos de Espiro/farmacologia , Compostos de Espiro/toxicidade , Streptomyces/metabolismo , Glutationa Transferase/metabolismo , Espécies Introduzidas , Acetaldeído/análogos & derivados , Álcoois GraxosRESUMO
Cisplatin-based chemotherapy is the mainstay in the treatment of germ cell tumors (GCTs). Glutathione S-transferases (GSTs) are polymorphic enzymes that catalyze the glutathione conjugation of alkylating agents, platinum compounds, and free radicals formed by chemotherapy and are thus implicated in developing treatment resistance. This study aimed to assess the expression level of GST mu 1 (GSTM1) and its association with treatment outcomes in patients with GCT. This translational study included tumor specimens from 207 patients with newly diagnosed GCTs, as well as cisplatin-sensitive GCT cell line xenografts and their resistant variants for all histological variants of GCTs. GSTM1 expression was detected by reverse transcription-quantitative PCR and immunohistochemistry using monoclonal antibodies, scored by the multiplicative quickscore (QS) method. GSTM1 expression was correlated with patient/tumor characteristics and treatment outcomes. The highest GSTM1 expression was observed in seminoma, followed by choriocarcinoma, embryonal carcinoma, and yolk sac tumor, while the lowest was observed in teratoma (p<0.0001). There was no association between GSTM1 expression in tumor tissue and patient/tumor characteristics. The low GSTM1 expression was associated with significantly better relapse-free survival compared with high GSTM1 (HR=0.50, 95% CI 0.23-1.09, p=0.03) but not overall survival (HR=0.61, 95% CI 0.24-1.54, p=0.22). Multivariate analysis showed that the prognostic value of GSTM1 was independent of the International Germ Cell Cancer Collaborative Group (IGCCCG) score. These data revealed the prognostic value of GSTM1 in GCTs, with a high GSTM1 expression associated with worse outcomes, suggesting that GSTM1 could be responsible, in part, for treatment resistance in GCTs.
Assuntos
Glutationa Transferase , Neoplasias Embrionárias de Células Germinativas , Humanos , Neoplasias Embrionárias de Células Germinativas/tratamento farmacológico , Neoplasias Embrionárias de Células Germinativas/patologia , Neoplasias Embrionárias de Células Germinativas/metabolismo , Neoplasias Embrionárias de Células Germinativas/mortalidade , Glutationa Transferase/metabolismo , Glutationa Transferase/genética , Masculino , Adulto , Cisplatino/uso terapêutico , Resultado do Tratamento , Adulto Jovem , Pessoa de Meia-Idade , Resistencia a Medicamentos Antineoplásicos , Neoplasias Testiculares/tratamento farmacológico , Neoplasias Testiculares/patologia , Neoplasias Testiculares/metabolismo , Neoplasias Testiculares/mortalidade , Adolescente , Animais , Feminino , Prognóstico , Linhagem Celular Tumoral , CamundongosRESUMO
BACKGROUND: Cancer initiation has long been "unknowable" in biology and medicine. In 1987, however, Moore and our research group observed single hepatocytes and minifoci that were strongly positive for glutathione S-transferase P-form (GST-P) in the rat liver as early as 2 to 3 days after initiation by diethylnitrosamine prior to the induction of GST-P+ foci and nodules. The induction of GST-P+ single hepatocytes, precursors of GST-P+ foci and nodules, was considered genetic. But, the details of the induction mechanism have remained unclear despite various examinations over a long period. METHODS: Male Sprague-Dawley rats (aged 6 weeks) were fed a basal diet containing either benzyl isothiocyanate (BITC, 0.5% by wt) or 2-acetylaminofluorene (AAF, 0.04%) ad libitum for appropriate time intervals. All animals were anesthetized and euthanized. The livers obtained were excised, cut into 3- to 4-mm-thick slices and fixed in cold acetone at 4 °C. The liver specimens were then sliced into 25-µm-thick sections in PBS using an automated microtome (Vibratome 1500 Sectioning System, Vibratome Products, NY, USA). Immunocytochemical staining was performed in free solution, and the results were examined via digital light microscopy (Coolscope, Nikon, Tokyo). RESULTS: 3D analysis using a vibratome showed that GST-P is rapidly excreted into the bile of the liver of animals in response to strong carcinogenic stress caused by promoters or initiators. "Rapid biliary excretion of GST-P" was widely and commonly observed in all hepatocytes, GST-P+ single hepatocytes, minifoci, foci and nodules under appropriate conditions. Surprisingly, on the basis of these key findings, a new mechanism of cancer initiation involving the transformation of hepatocytes into GST-P+ single hepatocytes and minifoci in animal livers was identified. In addition, the initiation process was determined to be nongenetic because mutation is an invisible rare event. CONCLUSIONS: This short review describes several details about breakthrough findings on cancer initiation in rat livers, the application of 3D analysis to other cancers and the importance in the genetic analysis in malignant diseases.
Assuntos
Hepatócitos , Fígado , Lesões Pré-Cancerosas , Animais , Hepatócitos/metabolismo , Masculino , Ratos , Fígado/patologia , Fígado/metabolismo , Lesões Pré-Cancerosas/patologia , Lesões Pré-Cancerosas/metabolismo , Lesões Pré-Cancerosas/induzido quimicamente , Ratos Sprague-Dawley , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/genética , Glutationa S-Transferase pi/metabolismo , Glutationa S-Transferase pi/genética , 2-Acetilaminofluoreno , Imageamento Tridimensional , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/induzido quimicamente , Neoplasias Hepáticas/genética , Carcinógenos/toxicidade , Glutationa Transferase/metabolismo , Glutationa Transferase/genéticaRESUMO
The pathogenesis of most cases of carpal tunnel syndrome is not clearly defined. There are some aspects of the disease that suggest a potential effect of genetic predispositions. Mutations (variants) within the genes encoding various subtypes of collagen synthesis, oligomerisation in the endoplasmic reticulum and inactivation of reactive oxygen species may be involved in the development of carpal tunnel syndrome. The objective of this study was to determine the role of DNA alterations within the COL11A, COL1A, COL5A1, COMP and GSTM1 genes in the pathogenesis of carpal tunnel syndrome based on a Polish population. STUDY DESIGN: In the discovery phase, a total of 96 patients with familial aggregation of CTS were genotyped using a Next Generation Sequencing panel in order to find possible mutations within the studied genes. The potential pathogenicity of the detected variants was investigated using the predictions of several in-silico algorithms and the TaqMan technology. In the association phase of the study, a group of 345 CTS patients and 1035 healthy controls were genotyped. RESULTS: A total of 35 splice-site or exonic non-synonymous variants were detected by NGS. We did not identify any clearly pathogenic or likely pathogenic alternations. The 30 variants were identified as benign or likely benign. Five missense changes were predicted as VUS and selected for association study. The COL5A1 c.1595 C>T (p.Ala532Val) was detected in one out of 345 cases and three out of 1035 controls (P=1, OR=1); this indicates that the variant is a neutral alteration. Four remaining variants - c.2840 C>A, c.5395 G>A, c.1331 C>G, c.1590 C>A - were present in none out of the 345 CTS patients and none out of 1035 controls. CONCLUSION: The main finding of this study was that there was no independent association between the variants of five examined genes and carpal tunnel syndrome. Four uncertain variants were identified that seem to be extremely rare in the Polish population.
Assuntos
Síndrome do Túnel Carpal , Cadeia alfa 1 do Colágeno Tipo I , Colágeno Tipo I , Colágeno Tipo V , Colágeno Tipo XI , Predisposição Genética para Doença , Glutationa Transferase , Humanos , Síndrome do Túnel Carpal/genética , Feminino , Masculino , Pessoa de Meia-Idade , Colágeno Tipo I/genética , Colágeno Tipo XI/genética , Colágeno Tipo V/genética , Glutationa Transferase/genética , Cadeia alfa 1 do Colágeno Tipo I/genética , Adulto , Genótipo , Idoso , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
Salvia leucantha is a perennial herb of the genus Salvia in the family Labiatae, which has a wide range of biological activities, mainly including inhibition of acetylcholinesterase, antibacterial, and anti-inflammatory activity. To explore the protective effects and mechanism of action of S. leucantha on Alzheimer's disease (AD), the anti-AD activity of SLE (extracts of S. leucantha) was determined by using a transgenic Caenorhabditis elegans (C. elegans) model (CL4176). Analyses included paralysis assay, phenotypic experiments, transcriptome sequencing, RNA interference (RNAi), heat shock assays, and gas chromatography-mass spectrometry (GC-MS). SLPE (S. leucantha petroleum ether extract) could significantly delay CL4176 paralysis and extend the longevity of C. elegans N2 without harmful effects. A total of 927 genes were significantly changed by SLPE treatment in C. elegans, mainly involving longevity regulatory pathways-nematodes, drug metabolism-cytochrome P450, and glutathione metabolic pathways. RNAi showed that SLPE exerted its anti-AD activity through up-regulation of the gene gst-5; the most abundant compound in SLPE analyzed by GC-MS was 2,4-Di-tert-butylphenol (2,4-DTBP), and the compound delayed nematode paralysis. The present study suggests that active components in S. leucantha may serve as new-type anti-AD candidates and provide some insights into their biological functions.
Assuntos
Doença de Alzheimer , Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Modelos Animais de Doenças , Extratos Vegetais , Animais , Caenorhabditis elegans/genética , Caenorhabditis elegans/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Doença de Alzheimer/genética , Doença de Alzheimer/metabolismo , Doença de Alzheimer/tratamento farmacológico , Regulação para Cima/efeitos dos fármacos , Salvia/química , Longevidade/genética , Longevidade/efeitos dos fármacos , Animais Geneticamente Modificados , Interferência de RNA , Glutationa Transferase/genética , Glutationa Transferase/metabolismoRESUMO
Plant glutathione transferases (GSTs) constitute a large and diverse family of enzymes that are involved in plant stress response, metabolism and defence, yet their physiological functions remain largely elusive. Consistent with the traditional view on GSTs across organisms as detoxification enzymes, in vitro most plant GSTs catalyse glutathionylation, conjugation of the tripeptide glutathione (GSH; γ-Glu-Cys-Gly) onto reactive molecules. However, when it comes to elucidating GST functions, it remains a key challenge that the endogenous plant glutathione conjugates (GS-conjugates) that would result from such glutathionylation reactions are rarely reported. Furthermore, GSTs often display high substrate promiscuity, and their proposed substrates are prone to spontaneous chemical reactions with GSH; hence, single-gene knockouts rarely provide clear chemotypes or phenotypes. In a few cases, GS-conjugates are demonstrated to be biosynthetic intermediates that are rapidly further metabolized towards a pathway end product, explaining their low abundance and rare detection. In this review, we summarize the current knowledge of plant GST functions and how and possibly why evolution has resulted in a broad and extensive expansion of the plant GST family. Finally, we demonstrate that endogenous GS-conjugates are more prevalent in plants than assumed and suggest they are overlooked as clues towards the identification of plant GST functions. This article is part of the theme issue 'The evolution of plant metabolism'.
Assuntos
Glutationa Transferase , Glutationa , Plantas , Glutationa Transferase/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/química , Glutationa/metabolismo , Plantas/enzimologia , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genéticaRESUMO
AIMS: This study identifies a unique glutathione S-transferase (GST) in extremophiles using genome, phylogeny, bioinformatics, functional characterization, and RNA sequencing analysis. METHODS AND RESULTS: Five putative GSTs (H0647, H0729, H1478, H3557, and H3594) were identified in Halothece sp. PCC7418. Phylogenetic analysis suggested that H0647, H1478, H0729, H3557, and H3594 are distinct GST classes. Of these, H0729 was classified as an iota-class GST, encoding a high molecular mass GST protein with remarkable features. The protein secondary structure of H0729 revealed the presence of a glutaredoxin (Grx) Cys-Pro-Tyr-Cys (C-P-Y-C) motif that overlaps with the N-terminal domain and harbors a topology similar to the thioredoxin (Trx) fold. Interestingly, recombinant H0729 exhibited a high catalytic efficiency for both glutathione (GSH) and 1-chloro-2, 4-dinitrobenzene (CDNB), with catalytic efficiencies that were 155- and 32-fold higher, respectively, compared to recombinant H3557. Lastly, the Halothece gene expression profiles suggested that antioxidant and phase II detoxification encoding genes are crucial in response to salt stress. CONCLUSION: Iota-class GST was identified in cyanobacteria. This GST exhibited a high catalytic efficiency toward xenobiotic substrates. Our findings shed light on a diversified evolution of GST in cyanobacteria and provide functional dynamics of the genes encoding the enzymatic antioxidant and detoxification systems under abiotic stresses.
Assuntos
Cianobactérias , Glutationa Transferase , Filogenia , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Glutationa Transferase/química , Cianobactérias/genética , Cianobactérias/enzimologia , Cianobactérias/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/química , Glutationa/metabolismo , Sequência de Aminoácidos , Glutarredoxinas/genética , Glutarredoxinas/metabolismo , Glutarredoxinas/químicaRESUMO
Sand flies infect more than 1 million people annually with Leishmania parasites and other bacterial and viral pathogens. Progress in understanding sand fly adaptations to xenobiotics has been hampered by the limited availability of genomic resources. To address this gap, we sequenced, assembled, and annotated the transcriptomes of 11 phlebotomine sand fly species. Subsequently, we leveraged these genomic resources to generate novel evolutionary insights pertaining to their adaptations to xenobiotics, including those contributing to insecticide resistance. Specifically, we annotated over 2,700 sand fly detoxification genes and conducted large-scale phylogenetic comparisons to uncover the evolutionary dynamics of the five major detoxification gene families: cytochrome P450s (CYPs), glutathione-S-transferases (GSTs), UDP-glycosyltransferases (UGTs), carboxyl/cholinesterases (CCEs), and ATP-binding cassette (ABC) transporters. Using this comparative approach, we show that sand flies have evolved diverse CYP and GST gene repertoires, with notable lineage-specific expansions in gene groups evolutionarily related to known xenobiotic metabolizers. Furthermore, we show that sand flies have conserved orthologs of (i) CYP4G genes involved in cuticular hydrocarbon biosynthesis, (ii) ABCB genes involved in xenobiotic toxicity, and (iii) two primary insecticide targets, acetylcholinesterase-1 (Ace1) and voltage gated sodium channel (VGSC). The biological insights and genomic resources produced in this study provide a foundation for generating and testing hypotheses regarding the molecular mechanisms underlying sand fly adaptations to xenobiotics.
Assuntos
Evolução Molecular , Resistência a Inseticidas , Inseticidas , Filogenia , Psychodidae , Animais , Psychodidae/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Genômica , Inativação Metabólica/genética , Xenobióticos/metabolismoRESUMO
Spodoptera litura is a significant agricultural pest, and its glutathione S-transferase (GST) plays a crucial role in insecticide resistance. This study aimed to investigate the relationship between the SlGSTe11 gene of S. litura and resistance to cyantraniliprole and nicotine. Transcriptome analysis revealed that SlGSTe11 is highly expressed mainly in fat bodies, with a significant increase in SlGSTe11 gene expression under induction by cyantraniliprole and nicotine. The ectopic expression of the SlGSTe11 gene in transgenic fruit flies resulted in a 5.22-fold increase in the tolerance to cyantraniliprole. Moreover, compared to the UAS-SlGSTe11 line, the Act5C-UAS>SlGSTe11 line laid more eggs and had a lower mortality after nicotine exposure. RNAi-mediated inhibition of SlGSTe11 gene expression led to a significant increase in the mortality of S. litura under cyantraniliprole exposure. In vitro metabolism experiments demonstrated that the recombinant SlGSTe11 protein efficiently metabolizes cyantraniliprole. Molecular docking results indicated that SlGSTe11 has a strong affinity for both cyantraniliprole and nicotine. These findings suggest that SlGSTe11 is involved in the development of resistance to cyantraniliprole and nicotine in S. litura.
Assuntos
Corpo Adiposo , Glutationa Transferase , Proteínas de Insetos , Resistência a Inseticidas , Inseticidas , Nicotina , Pirazóis , Spodoptera , ortoaminobenzoatos , Animais , Spodoptera/genética , Spodoptera/efeitos dos fármacos , Spodoptera/metabolismo , Spodoptera/enzimologia , Spodoptera/crescimento & desenvolvimento , Inseticidas/farmacologia , Inseticidas/metabolismo , Inseticidas/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Proteínas de Insetos/química , ortoaminobenzoatos/metabolismo , ortoaminobenzoatos/farmacologia , Pirazóis/farmacologia , Nicotina/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Glutationa Transferase/química , Resistência a Inseticidas/genética , Corpo Adiposo/metabolismo , Corpo Adiposo/enzimologia , Corpo Adiposo/efeitos dos fármacos , Simulação de Acoplamento MolecularRESUMO
Agricultural activities contribute to plastic pollution, with unintentional introduction and intentional use of plastic mulch films leading to the accumulation of microplastic particles in soils. The lack of removal techniques and scarce information on the effects on soil organisms, especially for biodegradable mulch films, necessitate an assessment of potential effects. This study aimed to elucidate the effects of mulch film microplastic on soil fauna by investigating reproduction output and subcellular responses before and after recovery from exposure. Two common soil organisms, Folsomia candida and Eisenia fetida, were exposed to petroleum-based polyethylene (PE) and biodegradable polylactic acid/polybutylene adipate terephthalate (PLA/PBAT) microplastic for 28 days, according to OECD guidelines 232 and 222, respectively. Juvenile numbers revealed no polymer- or concentration-dependent effects on E. fetida and F. candida reproduction after exposure to up to 5 and 10 g/kgdw soil, respectively. To provide a more sensitive and early indication of sublethal effects, subcellular responses in E. fetida were analyzed. Glutathione S-transferase (GST) activity increased with rising microplastic concentration; however, catalase (CAT), acetylcholine esterase (AChE) activity, and reactive oxygen species (ROS) did not differ from control levels. Further, the more environmentally relevant PE polymer was chosen for in-depth assessment of subcellular response after 28-day microplastic exposure and subsequent 28 days in uncontaminated soil with E. fetida. No significant differences in biomarker activity and stress levels were observed. We conclude that mulch film-derived microplastic did not adversely affect earthworm and collembolan species in this scenario, except for a slight induction in the detoxification enzyme glutathione S-transferase.
Assuntos
Microplásticos , Poluentes do Solo , Poluentes do Solo/toxicidade , Poluentes do Solo/análise , Animais , Microplásticos/toxicidade , Oligoquetos , Solo/química , Monitoramento Ambiental , Poliésteres , Polietileno , Plásticos , Glutationa Transferase/metabolismo , Artrópodes/efeitos dos fármacosRESUMO
During their development, amphibians undergo various physiological processes that may affect their susceptibility to environmental pollutants. Naturally occurring fluctuations caused by developmental events are often overlooked in ecotoxicological studies. Our aim is to investigate how biomarkers of oxidative stress are modulated at different stages of larval development in the Amazonian amphibian species, Physalaemus ephippifer. The premetamorphosis, prometamorphosis and metamorphic climax stages were used to analyze total antioxidant capacity (ACAP), glutathione S-transferase (GST) activity, lipid peroxidation (LPO) levels and the expression of genes nrf2, gst, gsr (glutathione reductase) and gclc (glycine-cysteine ligase, catalytic subunit). Although there was no difference in ACAP and the genes expression among the studied stages, individuals from the premetamorphosis and prometamorphosis showed higher GST activity than ones under the climax. LPO levels were highest in individuals from the metamorphic climax. The present study suggests that the oxidative status changes during ontogeny of P. ephippifer tadpoles, especially during the metamorphic climax, the most demanding developmental phase. Variations in the redox balance at different developmental stages may lead to a divergent response to pollution. Therefore, we recommend that studies using anuran larvae as biomonitors consider possible physiological differences during ontogeny in their respective analyses.
Assuntos
Anuros , Glutationa Transferase , Larva , Peroxidação de Lipídeos , Oxirredução , Estresse Oxidativo , Animais , Anuros/metabolismo , Anuros/crescimento & desenvolvimento , Larva/metabolismo , Larva/crescimento & desenvolvimento , Glutationa Transferase/metabolismo , Antioxidantes/metabolismo , Metamorfose Biológica , Biomarcadores/metabolismoRESUMO
OBJECTIVE: Aim: The objective of the research was to conduct a comprehensive longitudinal analysis of the temporal dynamics of glutathione system functionality in individuals diagnosed with paranoid schizophrenia. Specifically, the research was focused on investigating variations in the profiles of glutathione-dependent enzymes, with meticulous consideration given to the duration of the illness. PATIENTS AND METHODS: Materials and Methods: The study group comprised 300 individuals officially diagnosed with 'Paranoid Schizophrenia,' subdivided into five subgroups, each consisting of 60 patients. The subgroups were defined as follows: Subgroup I included 60 patients with a disease duration ranging from 3 to 5 years; Subgroup II comprised 60 patients with a duration of 6 to 10 years; Subgroup III consisted of 60 patients with a duration of 11 to 15 years; Subgroup IV included 60 patients with a duration of 16 to 20 years; and Subgroup V encompassed 60 patients with a duration of 21 years and older. The comparison group comprised 20 patients diagnosed with "Primary psychotic episode". RESULTS: Results: The research demonstrates a consistent and noteworthy reduction in the enzymatic activities of glutathione peroxidase, glutathione reductase, and glutathione-S-transferase in various Subgroups of paranoid schizophrenia patients. The observed declines are particularly prominent within the first 3-5 years of the illness, show casing statistically significant reductions. Patients with prolonged illness durations, especially surpassing 21 years, display substantial reductions in all three enzymes, suggesting a cumulative enzymatic impact associated with prolonged illness. CONCLUSION: Conclusions: The identification of critical periods of inhibition in the glutathione protection chain, provides valuable information about potential therapeutic interventions for individuals with paranoid schizophrenia.
Assuntos
Esquizofrenia Paranoide , Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Glutationa Redutase/sangue , Glutationa Transferase/metabolismo , Adulto Jovem , Estudos Longitudinais , Fatores de Tempo , Glutationa/metabolismoRESUMO
The fall armyworm (FAW), Spodoptera frugiperda Smith (Lepidoptera: Noctuidae), poses a significant threat to food security, necessitating effective management strategies. While chemical control remains a primary approach, understanding the toxicity and detoxification mechanisms of different insecticides is crucial. In this study, we conducted leaf-dipping bioassays to assess the toxicity of quinalphos and beta-cypermethrin·emamectin benzoate (ß-cyp·EMB) on S. frugiperda larvae. Additionally, we assessed the response of alterations in CarE, GST, MFO, and AChE activities to sublethal concentrations of these insecticides over various treatment durations. Results indicated that ß-cyp·EMB exhibited higher toxicity than quinalphos in S. frugiperda. Interestingly, the highest activities of GST, CarE, MFO, and AChE were observed at 6â¯h exposure to LC10 and LC25 of ß-cyp·EMB, surpassing equivalent sublethal concentrations of quinalphos. Subsequently, GST and CarE activities exposure to ß-cyp·EMB steadily decreased, while MFO and AChE activities exposure to both insecticides was initially decreased then increased. Conversely, two sublethal concentrations of quinalphos notably enhanced GST activity across all exposure durations, with significantly higher than ß-cyp·EMB at 12-48â¯h. Similarly, CarE activity was also increased at various durations. Our research has exhibited significant alterations in enzyme activities exposure to both concentration and duration. Furthermore, Pearson correlation analysis showed significant correlations among these enzyme activities at different treatment durations. These findings contribute to a better understanding of detoxification mechanisms across different insecticides, providing valuable insights for the rational management of S. frugiperda populations.
Assuntos
Inativação Metabólica , Inseticidas , Ivermectina , Larva , Piretrinas , Spodoptera , Animais , Inseticidas/toxicidade , Spodoptera/efeitos dos fármacos , Ivermectina/análogos & derivados , Ivermectina/toxicidade , Larva/efeitos dos fármacos , Piretrinas/toxicidade , Acetilcolinesterase/metabolismo , Glutationa Transferase/metabolismoRESUMO
Schistosomiasis caused by Schistosoma japonicum (S. japonicum) is a major public health problem in the Philippines, China and Indonesia. In this study, the immunopotentiator CpG-ODN was encapsulated within chitosan nanoparticles (Chi NPs) to create a combination adjuvant (Chi-CpG NP). This approach was employed to enhance the immunogenicity of 26 kDa glutathione S-transferase (Sj26GST) from S. japonicum through intranasal immunization. The results demonstrated higher levels of specific anti-Sj26GST antibodies and Sj26GST-specific splenocyte proliferation compared to mice that were immunized with Sj26GST + Chi-CpG NP. Cytokine analysis of splenocytes revealed that the Sj26GST + Chi-CpG NP induced a slight Th1-biased immune response, with increased production of IFN-γ by CD4+ T-cells in the spleen. Subsequently, mice were intradermally inoculated with 1 × 107 organisms in the Coeliac cavity. The bacterial organ burden detected in the liver of immunized mice suggested that Sj26GST + Chi-CpG NP enhances protective immunity to inhibit S. japonicum colonization. Therefore, Sj26GST + Chi-CpG NP vaccination enhances Sj26GST-specific immunogenicity and provides protection against S. japonicum.