RESUMEN
Under nitrogen deprivation (-N), cyanobacterium Synechocystis sp. PCC 6803 exhibits growth arrest, reduced protein content, and remarkably increased glycogen accumulation. However, producing glycogen under this condition requires a two-step process with cell transfer from normal to -N medium. Metabolic engineering and chemical treatment for rapid glycogen accumulation can bypass the need for two-step cultivation. For example, recent studies indicate that individually disrupting hydrogen (H2) or poly(3-hydroxybutyrate) (PHB) synthesis, or treatment with methyl viologen (MV), effectively increases glycogen accumulation in Synechocystis. Here we explore the effects of disrupted H2 or poly(3-hydroxybutyrate) synthesis, together with MV treatment to on enhanced glycogen accumulation in Synechocystis grown in normal medium. Wild-type cells without MV treatment exhibited low glycogen content of less than 6% w/w dry weight (DW). Compared with wild type, disrupting PHB synthesis combined with MV treatment did not increase glycogen content. Disrupted H2 production without MV treatment yielded up to 11% w/w DW glycogen content. Interestingly, when combined, disrupted H2 production with MV treatment synergistically enhanced glycogen accumulation to 51% and 59% w/w DW within 3 and 7 days, respectively. Metabolomic analysis suggests that MV treatment mediated the conversion of proteins into glycogen. Metabolomic and transcriptional-expression analysis suggests that disrupted H2 synthesis under MV treatment positively influenced glycogen synthesis. Disrupted H2 synthesis under MV treatment significantly increased NADPH levels. This increased NADPH content potentially contributed to the observed enhancements in antioxidant activity against MV-induced oxidants, O2 evolution, and metabolite substrates levels for glycogen synthesis in normal medium, ultimately leading to enhanced glycogen accumulation in Synechocystis. KEY MESSAGE: Combining disrupted hydrogen-gas synthesis and the treatment by photosynthesis electron-transport inhibitor significantly enhance glycogen production in cyanobacteria.
Asunto(s)
Glucógeno , Hidrógeno , Paraquat , Fotosíntesis , Synechocystis , Glucógeno/metabolismo , Synechocystis/metabolismo , Synechocystis/efectos de los fármacos , Synechocystis/genética , Fotosíntesis/efectos de los fármacos , Hidrógeno/metabolismo , Paraquat/farmacología , Hidroxibutiratos/metabolismo , Transporte de Electrón/efectos de los fármacos , Poliésteres/metabolismo , PolihidroxibutiratosRESUMEN
BACKGROUND: Paraquat (PQ) is a widely used and highly toxic herbicide that poses a significant risk to human health. The main consequence of PQ poisoning is pulmonary fibrosis, which can result in respiratory failure and potentially death. Our research aims to uncover a crucial mechanism in which PQ poisoning induces senescence in epithelial cells, ultimately regulating the activation of pulmonary fibroblasts through the exosomal pathway. METHODS: Cellular senescence was determined by immunohistochemistry and SA-ß-Gal staining. The expression of miRNAs was measured by qPCR. Pulmonary fibroblasts treated with specific siRNA of SIRT1 or LV-SIRT1 were used to analysis senescent exosomes-mediated fibroblasts activation. Luciferase reporter assay and western blot were performed to elucidated the underlying molecular mechanisms. The effects of miR-217-5p antagomir on pulmonary fibrosis were assessed in PQ-poisoned mice models. RESULTS: Impairing the secretion of exosomes effectively mitigates the harmful effects of senescent epithelial cells on pulmonary fibroblasts, offering protection against PQ-induced pulmonary fibrosis in mice. Additionally, we have identified a remarkable elevation of miR-217-5p expression in the exosomes of PQ-treated epithelial cells, which specifically contributes to fibroblasts activation via targeted inhibition of SIRT1, a protein involved in cellular stress response. Remarkably, suppression of miR-217-5p effectively impaired senescent epithelial cells-induced fibroblasts activation. Further investigation has revealed that miR-217-5p attenuated SIRT1 expression and subsequently resulted in enhanced acetylation of ß-catenin and Wnt signaling activation. CONCLUSION: These findings highlight a potential strategy for the treatment of pulmonary fibrosis induced by PQ poisoning. Disrupting the communication between senescent epithelial cells and pulmonary fibroblasts, particularly by targeting the miR-217-5p/SIRT1/ß-catenin axis, may be able to alleviate the effects of PQ poisoning on the lungs.
Asunto(s)
Exosomas , MicroARNs , Fibrosis Pulmonar , Humanos , Ratones , Animales , Fibrosis Pulmonar/genética , Paraquat/metabolismo , Paraquat/farmacología , beta Catenina/metabolismo , Exosomas/metabolismo , Sirtuina 1/metabolismo , Pulmón/patología , MicroARNs/genética , Células Epiteliales/patología , Fibroblastos/metabolismoRESUMEN
Plants, being sessile organisms, constantly need to respond to environmental stresses, often leading to the accumulation of reactive oxygen species (ROS). While ROS can be harmful, they also act as second messengers guiding plant growth and stress responses. Because chloroplasts are sensitive to environmental changes and are both a source and a target of ROS during stress conditions, they are important in conveying environmental changes to the nucleus, where acclimation responses are coordinated to maintain organellar and overall cellular homeostasis. ANAC102 has previously been established as a regulator of ß-cyclocitral-mediated chloroplast-to-nucleus signaling, protecting plants against photooxidative stress. However, debates persist about where ANAC102 is located-in chloroplasts or in the nucleus. Our study, utilizing the genomic ANAC102 sequence driven by its native promoter, establishes ANAC102 primarily as a nuclear protein, lacking a complete N-terminal chloroplast-targeting peptide. Moreover, our research reveals the sensitivity of plants overexpressing ANAC102 to severe superoxide-induced chloroplast oxidative stress. Transcriptome analysis unraveled a dual role of ANAC102 in negatively and positively regulating genome-wide transcriptional responses to chloroplast oxidative stress. Through the integration of published data and our own study, we constructed a comprehensive transcriptional network, which suggests that ANAC102 exerts direct and indirect control over transcriptional responses through downstream transcription factor networks, providing deeper insights into the ANAC102-mediated regulatory landscape during oxidative stress.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Estrés Oxidativo , Paraquat , Factores de Transcripción , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/fisiología , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Paraquat/farmacología , Proteínas Nucleares/metabolismo , Proteínas Nucleares/genética , Regulación de la Expresión Génica de las Plantas , Cloroplastos/metabolismoRESUMEN
Paraquat (1,1'-dimethyl-4,4'-bipyridyl dichloride) is an herbicide widely used worldwide and officially banned in Brazil in 2020. Kidney lesions frequently occur, leading to acute kidney injury (AKI) due to exacerbated reactive O2 species (ROS) production. However, the consequences of ROS exposure on ionic transport and the regulator local renin-angiotensin-aldosterone system (RAAS) still need to be elucidated at a molecular level. This study evaluated how ROS acutely influences Na+-transporting ATPases and the renal RAAS. Adult male Wistar rats received paraquat (20 mg/kg; ip). After 24 h, we observed body weight loss and elevation of urinary flow and serum creatinine. In the renal cortex, paraquat increased ROS levels, NADPH oxidase and (Na++K+)ATPase activities, angiotensin II-type 1 receptors, tumor necrosis factor-α (TNF-α), and interleukin-6. In the medulla, paraquat increased ROS levels and NADPH oxidase activity but inhibited (Na++K+)ATPase. Paraquat induced opposite effects on the ouabain-resistant Na+-ATPase in the cortex (decrease) and medulla (increase). These alterations, except for increased serum creatinine and renal levels of TNF-α and interleukin-6, were prevented by 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (tempol; 1 mmol/L in drinking water), a stable antioxidant. In summary, after paraquat poisoning, ROS production culminated with impaired medullary function, urinary fluid loss, and disruption of Na+-transporting ATPases and angiotensin II signaling.
Asunto(s)
Paraquat , Sistema Renina-Angiotensina , Ratas , Animales , Masculino , Especies Reactivas de Oxígeno/metabolismo , Paraquat/metabolismo , Paraquat/farmacología , Angiotensina II/metabolismo , Angiotensina II/farmacología , Creatinina/metabolismo , Creatinina/orina , Interleucina-6 , Factor de Necrosis Tumoral alfa/metabolismo , Ratas Wistar , Riñón , Adenosina Trifosfatasas/metabolismo , Adenosina Trifosfatasas/farmacología , Sodio/metabolismo , Sodio/farmacología , NADPH Oxidasas/metabolismo , NADPH Oxidasas/farmacologíaRESUMEN
While the spatial distribution pattern of fish is increasingly used for toxicological test of chemicals or wastewater, no ideal parameter is available for quantitative assessment of spatial distribution, especially uneven distribution with multiple hotspots. Here, to develop a quantitative assessment parameter for spatial distribution, the zebrafish were exposed to ethanol, pentylenetetrazole (PTZ), paraquat dichloride (paraquat) and wastewater, followed by a behavioral test in a narrow tank. Behavioral data was acquired and analyzed by idTracker and MATLAB. By comparing the effects of all treatments on behavior parameters, we confirmed that the spatial distribution was more easily altered rather than general locomotor parameters, e.g. 0.7-70 mg/L PTZ and 5-20 mg/L paraquat being effective for altering spatial distribution but having little effects on general locomotor parameters. Based on the heatmap, i.e., the cumulative proportion of grids and that of frequency in grids, we calculated the behavioral Gini coefficient (Gb) for quantitative assessment of fish spatial distribution. The Gini coefficient ranged from zero to 1, with larger values meaning poorer evenness of spatial distribution. Of note, Gb showed smaller coefficient of variations (CV) with 3%-19% between replicate tanks in all treatments than the highest frequency (4%-79%), displaying well robustness. Especially, Gb addressed the challenge of the complicated heatmap with multiple hotspots. Overall, the behavioral Gini coefficient we established is an ideal parameter to quantitatively assess spatial distribution of fish shoal, which is expected to be applied in toxicity testing for chemicals and wastewater and automatic quality monitoring for surface water and aquaculture water.
Asunto(s)
Aguas Residuales , Pez Cebra , Animales , Paraquat/farmacología , Conducta Animal , AguaRESUMEN
Historically, males have frequently been portrayed as the manipulative and deceptive gender, while females are often seen as adopting a coy and passive role. In this context, it is proposed that males use a terminal investment strategy, misleading females about their true poor condition, while females passively opt to mate with these deceptive males. However, we hypothesize that females in suboptimal condition may also engage in a terminal investment strategy by mimicking or enhancing their attractiveness to match that of females in better conditions. We studied this hypothesis in Tenebrio molitor, by subjecting females to three varying doses of lipopolysaccharides of Escherichia coli (LPS; 0.25, 0.5, or 1 mg ml-1), or three doses of the pro-oxidant Paraquat (PQ; 20, 40 or 80 mM), and subsequently assessing their survival and attractiveness to males. The LPS treatments and 20 mM of PQ had no significant effect on the survival or attractiveness of the females. However, females treated with 40 or 80 mM PQ survived fewer days compared to the control group. Those injected with 40 mM were more attractive than their control counterparts, while those treated with 80 mM were less attractive. Since the identical doses of LPS, which induce terminal investment in males, had no effect on females, we suggest sexual dimorphism in terminal investment. Furthermore, similar to males, if the stressor reaches a sufficiently high level, the signal becomes honest. These findings highlight how the quantity of stressors influences support for the terminal investment strategy in both males and females. Notably, this study challenges prevailing notions regarding gender roles in sexual selection, indicating that females, not just males, conceal their poor condition to attract mating partners.
Asunto(s)
Lipopolisacáridos , Conducta Sexual Animal , Femenino , Masculino , Animales , Conducta Sexual Animal/efectos de los fármacos , Conducta Sexual Animal/fisiología , Lipopolisacáridos/farmacología , Tenebrio/fisiología , Tenebrio/efectos de los fármacos , Paraquat/farmacologíaRESUMEN
Methyl viologen (MV), also known as paraquat, is a widely used herbicide but has also been reported as highly toxic to different life forms. The mode of its operation is related to superoxide radical (O2.-) production and consequent oxidative damage. However, besides the damage to key macromolecules, reactive oxygen species (ROS; to which O2.- belongs) are also known as regulators of numerous ion transport systems located at cellular membranes. In this study, we used MV as a tool to probe the role of O2.- in regulating membrane-transport activity and systemic acquired tolerance in halophytic Chenopodium quinoa and glycophytic spinach plants. Both plant species showed growth reduction in terms of reduced shoot length, lower shoot fresh and dry weight, photosynthesis rate, and chlorophyll contents; however, quinoa showed less reduction in growth compared with spinach. This whole plant response was further examined by measuring the ion concentration, gene expression of ion transporters, activation of antioxidants, and osmolyte accumulation. We observed that at the mechanistic level, the differences in growth in response to MV were conferred by at least four complementary physiological mechanisms: (1) higher K+ loss from spinach leaves resulted from higher expression of MV-induced plasma membrane-based depolarization-activated K+ efflux GORK channel, (2) higher activation of high-affinity K+ uptake transporter HAK5 in quinoa, (3) higher antioxidant production and osmolyte accumulation in quinoa as compared with spinach, and (4) maintaining a higher rate of photosynthesis due to higher chlorophyll contents, and efficiency of photosystem II and reduced ROS and MDA contents. Obtained results also showed that MV induced O2.- significantly reduced N contents in both species but with more pronounced effects in glycophytic spinach. Taken together this study has shown the role of O2.- in regulating membrane ion transport and N metabolism in the leaves of halophyte vs. glycophyte in the context of oxidative stress tolerance.
Asunto(s)
Chenopodium quinoa , Homeostasis , Oxidación-Reducción , Fotosíntesis , Potasio , Spinacia oleracea , Superóxidos , Chenopodium quinoa/metabolismo , Spinacia oleracea/metabolismo , Spinacia oleracea/efectos de los fármacos , Superóxidos/metabolismo , Potasio/metabolismo , Clorofila/metabolismo , Paraquat/farmacología , Hojas de la Planta/metabolismo , Antioxidantes/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Objective: To investigate the effects of duration, temperature and shake on paraquat (PQ) concentration in the blood of PQ-exposed rats during the specinen preservation and transportation. Methods: In March 2021, 60 SD male rats of Specific Pathogen Free class were randomly divided into low-dose group (10 mg/kg PQ) and high-dose group (80 mg/kg PQ). Each group was divided into 5 subgroups (normal temperature group, cold storage group, 37 ℃ storage group, shaking on normal temperature group and shaking on 37 ℃ group), six rats in each subgroup. The rats were given intraperitoneal injection of PQ, 1 h after exposure, the blood samples were obtained by cardiac extraction. After different interventions, the concentrations of PQ were detected and compared before and after the intervention in each subgroup. Results: In the shaking on 37 ℃ group, the results of PQ concentrations in PQ-exposed rats were significantly lower than those before the intervention (P<0.05). In the other subgroups, the results were not significantly different compared with before intervention (P>0.05) . Conclusion: The concentration of PQ in the blood of rats exposed to PQ was decreased by shaking for 4 hours at 37 ℃.
Asunto(s)
Ratas , Masculino , Animales , Ratas Sprague-Dawley , Paraquat/farmacología , PulmónRESUMEN
The effectiveness of several sulfhydryl compounds in the treatment of paraquat intoxication has been previously tested based on their antioxidant ability. However, practical guidelines for their clinical use remain to be determined. As a preliminary pharmacokinetic study on sulfhydryl compounds, we attempted to establish the optimal concentration of N-acetyl-L-cysteine, glutathione, superoxide dismutase, and catalase. We measured the antioxidant effect of these antioxidants in normal pooled plasma and on intracellular reactive oxygen species (ROS) induced by paraquat. N-acetyl-L-cysteine begins to suppress the production of ROS in plasma at concentrations as low as 5 mM, with the suppression being maximal at 40 mM. In the same way, glutathione increased the total antioxidant status in plasma at concentrations of 5-40 mM in a dose-dependent manner. Complete suppression of ROS in plasma induced by exposure to 500 micrometer paraquat for 40 min was observed when using 40 mM N-acetyl-L-cysteine and 5 mM glutathione. These concentrations are comparable with 50 units of catalase, which reduced ROS at concentrations of 5-100 units. Further pharmacokinetic study into the systemic administration of these antioxidants is necessary, using effective concentrations of 5-40 mM for both N-acetyl-L-cysteine and glutathione, and 1-50 units of catalase.