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1.
Biomed Res Int ; 2021: 6678165, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33604382

RESUMO

Objective: We aimed to investigate the protective effect of s-nitrosoglutathione (SNG) pretreatment on acute kidney injury (AKI) in septic rats. Methods: We constructed a rat model of sepsis by cecal ligation and puncture and observed the survival of the rats. We obtained kidney and blood samples from rats, observed the pathological damage to the kidney tissues, and evaluated kidney function and the expression levels of inflammatory factors. We also detected the expression of induced nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the kidneys by immunohistochemistry and evaluated the apoptosis of kidney tubular epithelial cells (KTEC) by TUNEL. Results: Pretreatment with SNG significantly reduced the mortality of septic rats, attenuated kidney pathological damage, and decreased the levels of serum creatinine, plasma neutrophil gelatinase-associated lipocalin, and plasma kidney injury molecule-1. Moreover, SNG pretreatment decreased the levels of TNF-α and IL-1ß in serum and kidney and reduced the expressions of NO, iNOS, PGE2, and COX-2 in the kidneys. Furthermore, pretreatment with SNG significantly reduced the apoptotic rate of KTEC and decreased the levels of caspase-3 and Bax mRNA, but increased the level of Bcl-2 mRNA. Conclusion: Pretreatment with SNG has a protective effect on AKI in septic rats, and the specific mechanisms are related to inhibition of inflammation, oxidation, and apoptosis.


Assuntos
Injúria Renal Aguda , Apoptose/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , S-Nitrosoglutationa/farmacologia , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/patologia , Animais , Inflamação/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Sepse/metabolismo , Sepse/patologia
2.
Cell Prolif ; 54(3): e12990, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33458941

RESUMO

OBJECTIVES: S-nitrosoglutathione reductase (GSNOR), a protein denitrosylase, protects the mitochondria from mitochondrial nitrosative stress. Mammalian preimplantation embryos are mitochondria-rich, but the effects of GSNOR on mitochondrial function in preimplantation embryos are not well-studied. In the present study, we investigate whether GSNOR plays a role in mitochondrial regulation during porcine preimplantation embryo development. MATERIALS AND METHODS: GSNOR dsRNA was employed to knock down the expression of GSNOR, and Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME), a pan-NOS inhibitor, was used to prevent protein S-nitrosylation. Mitochondrial amount and function in embryo development were assessed by performing immunofluorescence staining, Western blot, fluorescent probe and real-time reverse transcription PCR. RESULTS: GSNOR knock-down significantly impaired blastocyst formation and quality and markedly induced the increase in protein S-nitrosylation. Notably, GSNOR knock-down-induced overproduction of S-nitrosylation caused mitochondrial dysfunction, including mitochondrial membrane potential depolarization, mitochondria-derived reactive oxygen species (ROS) increase and ATP deficiency. Interestingly, GSNOR knock-down-induced total mitochondrial amount increase, but the ratio of active mitochondria reduction, suggesting that the damaged mitochondria were accumulated and mitochondrial clearance was inhibited. In addition, damaged mitochondria produced more ROS, and caused DNA damage and apoptosis. Importantly, supplementation with L-NAME reverses the increase in S-nitrosylation, accumulation of damaged mitochondria, and oxidative stress-induced cell death. Interestingly, autophagy was downregulated after GSNOR knock-down, but reversed by L-NAME treatment. Thus, GSNOR maintains mitochondrial homeostasis by promoting autophagy and the clearing of damaged mitochondria in porcine preimplantation embryos.


Assuntos
Homeostase/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , S-Nitrosoglutationa/farmacologia , Aldeído Oxirredutases/efeitos dos fármacos , Animais , Autofagia/efeitos dos fármacos , Blastocisto/metabolismo , Mitocôndrias/metabolismo , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Oxirredutases/metabolismo , Suínos
3.
Int J Mol Sci ; 21(18)2020 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-32932738

RESUMO

We characterized modes of action of NO-donor S-nitrosoglutathione (GSNO) and NO-synthase inhibitor l-NAME derived from dicrotic (DiN) and anacrotic (AnN) notches of rat arterial pulse waveform (APW) in the condition of increased/decreased NO bioavailability. The cross-relationship patterns of DiN and AnN with 34 hemodynamic parameters (HPs) induced by GSNO and l-NAME are presented. After GSNO bolus administration, approximate non-hysteresis relationships were observed in the difference between DiN-AnN (mmHg) blood pressure (BP) and other 19 HPs, suggesting that these HPs, i.e., their signaling pathways, responding to NO concentration, are directly connected. Hysteresis relationships were observed between DiN-AnN (mmHg) and other 14 HPs, suggesting that signaling pathways of these HPs are indirectly connected. The hysteresis relationships were only observed between the time interval DiN-AnN (ms) and other 34 HPs, indicating no direct connection of signaling pathways. The cross-relationship patterns of DiN-AnN (mmHg), but not DiN-AnN (ms), induced by l-NAME were in accordance to the increased NO bioavailability induced by GSNO. In conclusion, we found the non-hysteresis/hysteresis cross-relationship "patterns" of DiN-AnN intervals to other HPs in the presence of GSNO that revealed their direct or indirect signaling pathways connections. This may contribute to our understanding of biological effects of natural substances that modulate NO production and/or NO signaling pathways.


Assuntos
Artérias/metabolismo , Artérias/fisiologia , Pressão Sanguínea/fisiologia , Sistema Cardiovascular/metabolismo , Sistema Cardiovascular/fisiopatologia , Óxido Nítrico/metabolismo , Animais , Artérias/efeitos dos fármacos , Disponibilidade Biológica , Pressão Sanguínea/efeitos dos fármacos , Sistema Cardiovascular/efeitos dos fármacos , Hemodinâmica/efeitos dos fármacos , Hemodinâmica/fisiologia , Masculino , NG-Nitroarginina Metil Éster/metabolismo , NG-Nitroarginina Metil Éster/farmacologia , Ratos , Ratos Wistar , S-Nitrosoglutationa/metabolismo , S-Nitrosoglutationa/farmacologia , Transdução de Sinais/fisiologia
4.
Nitric Oxide ; 104-105: 1-10, 2020 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-32771473

RESUMO

Nitric oxide (NO) deficiency is often associated with several acute and chronic diseases. NO donors and especially S-nitrosothiols such as S-nitrosoglutathione (GSNO) have been identified as promising therapeutic agents. Although their permeability through the intestinal barrier have recently be proved, suitable drug delivery systems have to be designed for their oral administration. This is especially challenging due to the physico-chemical features of these drugs: high hydrophilicity and high lability. In this paper, three types of particles were prepared with an Eudragit® polymer: nanoparticles and microparticles obtained with a water-in-oil-in-water emulsion/evaporation process versus microparticles obtained with a solid-in-oil-in-water emulsion/evaporation process. They had a similar encapsulation efficiency (around 30%), and could be freeze-dried then be stored at least one month without modification of their critical attributes (size and GSNO content). However, microparticles had a slightly slower in vitro release of GSNO than nanoparticles, and were able to boost by a factor of two the drug intestinal permeability (Caco-2 model). Altogether, this study brings new data about GSNO intestinal permeability and three ready-to-use formulations suitable for further preclinical studies with oral administration.


Assuntos
Portadores de Fármacos/química , Nanopartículas/química , Doadores de Óxido Nítrico/farmacologia , S-Nitrosoglutationa/farmacologia , Células CACO-2 , Portadores de Fármacos/toxicidade , Composição de Medicamentos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Liofilização , Humanos , Mucosa Intestinal/metabolismo , Nanopartículas/toxicidade , Óxido Nítrico/metabolismo , Doadores de Óxido Nítrico/química , Doadores de Óxido Nítrico/toxicidade , Tamanho da Partícula , S-Nitrosoglutationa/química , S-Nitrosoglutationa/toxicidade
5.
J Tissue Eng Regen Med ; 14(6): 807-818, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32330363

RESUMO

Mounting evidence showing that local nitric oxide (NO) delivery may significantly improve the wound healing process has stimulated the development of wound dressings capable of releasing NO topically. Herein, we describe the preparation of a self-expandable NO-releasing hydrolyzed collagen sponge (CS), charged with the endogenously found NO donor, S-nitrosoglutathione (GSNO). We show that cold pressed and GSNO-charged CS (CS/GSNO) undergo self-expansion to its original 3D shape upon water absorption to a swelling degree of 2,300 wt%, triggering the release of free NO. Topical application of compressed CS/GSNO on wounds in an animal model showed that exudate absorption by CS/GSNO leads to the release of higher NO doses during the inflammatory phase and progressively lower NO doses at later stages of the healing process. Moreover, treated animals showed significant increase in the mRNA expression levels of monocyte chemoattractant protein-1 (MCP-1), murine macrophage marker (F4/80), transforming growth factor beta (TGF-ß), stromal cell-derived factor 1 (SDF-1), insulin-like growth factor-1 (IGF-1), nitric oxide synthase(iNOS), and matrix metalloproteinase(MMP-9). Cluster differentiation 31 (CD31), vascular endothelial growth factor (VEGF), and F4/80 were measured on Days 7 and 12 by immunohistochemistry in the cicatricial tissue. These results indicate that the topical delivery of NO enhances the migration and infiltration of leucocytes, macrophages, and keratinocytes to the wounded tissue, as well as the neovascularization and collagen deposition, which are correlated with an accelerated wound closure. Thus, self-expandable CS/GSNO may represent a novel biocompatible and active wound dress for the topical delivery of NO on wounds.


Assuntos
Colágeno , Óxido Nítrico , S-Nitrosoglutationa , Cicatrização/efeitos dos fármacos , Ferimentos e Lesões , Animais , Colágeno/química , Colágeno/farmacologia , Modelos Animais de Doenças , Implantes de Medicamento/química , Implantes de Medicamento/farmacocinética , Implantes de Medicamento/farmacologia , Masculino , Camundongos , Óxido Nítrico/química , Óxido Nítrico/farmacocinética , Óxido Nítrico/farmacologia , S-Nitrosoglutationa/química , S-Nitrosoglutationa/farmacocinética , S-Nitrosoglutationa/farmacologia , Ferimentos e Lesões/tratamento farmacológico , Ferimentos e Lesões/metabolismo , Ferimentos e Lesões/patologia
6.
Plant Physiol Biochem ; 150: 204-208, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32155448

RESUMO

Nitric oxide (NO) and potassium (K+) exert a profound influence on the acclimation of plants to multiple stress conditions. A recent report indicated that exogenous addition of an NO donor causes, under conditions of adequate K+ supply, a detrimental effect on K+ status. It remains unknown whether an exogenous NO source could negatively affect the potential capture of this element when plants are faced with a K+ shortage. In this work we offer evidence that, under conditions of K+-deprivation, the addition of the naturally occurring NO donor, S-nitrosoglutathione (GSNO), diminishes the potential inward transport of the K+-analogue rubidium (Rb+) from diluted Rb+ concentrations in Arabidopsis thaliana. Studies with the akt1-2 mutant, lacking the AKT1 inward-rectifier K+-channel involved in K+-uptake, unveiled that the effect of GSNO on Rb+-influx involves a non-AKT1 component. In addition, exposure to the NO-donor led to down-regulation of transcripts coding for the AtHAK5 K+-transporter, a major component of the K+-transport machinery in K+-deprived plants. Moreover, studies with the hak5 mutant showed that GSNO could either stimulate Rb+-uptake or does not lead to a significant effect on Rb+-uptake relative to -K+ and to -K+ in the presence of decayed GSNO, respectively, thus indicating that the presence of AtHAK5 is required for GSNO exerting an inhibitory effect.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Rubídio , S-Nitrosoglutationa , Arabidopsis/efeitos dos fármacos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Doadores de Óxido Nítrico/farmacologia , Potássio/metabolismo , Rubídio/metabolismo , S-Nitrosoglutationa/farmacologia
7.
Exp Neurol ; 328: 113262, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32119935

RESUMO

BACKGROUND: S-Nitrosoglutathione (GSNO) is a nitric oxide donor that has been investigated for neuroprotective and neuro-recovery effect. We aimed to conduct a systematic review on the published literatures using GSNO in both pre-clinical and clinical stroke studies. METHODS: We searched PubMed up to June 30, 2019, using the following keywords: S-Nitrosoglutathione, GSNO, stroke, cerebrovascular, carotid arteries, middle cerebral artery, and middle cerebral artery occlusion. Only studies published in the English language providing efficacy results of GSNO on ischemic stroke were included. Stroke Therapy Academic Industry Roundtable (STAIR) score was used to assess the quality of pre-clinical studies and PEDro score for clinical trials. A meta-analysis was conducted to compare the effect size. RESULTS: Of 39 articles identified, 10 (6 for pre-clinical and 4 for clinical studies) met the eligibility criteria and were included. The median STAIR score across the pre-clinical studies was 5.5 (range: 4-7), and the median PEDro score for the 4 clinical trials was 10 (ranged: 6 to 10). Among the 6 pre-clinical studies, GSNO reduced infarct size in 6 studies and improved neurological behavior scales in 5 studies compared to placebo. Inverse-variance weighted linear meta-analysis of standardized mean difference (Hedge's g) on 4 human studies revealed a big effect size (Hedge's g = -0.82, 95% CI: [-1.26, -0.38], P = .0003) favoring the GSNO group in term of reducing embolic signals. I2 value was 0 across the included clinical studies in the meta-analysis. CONCLUSIONS: Pre-clinical studies showed positive benefit of GSNO in animal stroke models. The meta-analysis of clinical studies demonstrated that GSNO is effective in reducing embolic signals in patients with symptomatic internal carotid artery stenosis undergoing carotid endarterectomy or stenting. Further investigation of this molecule is warranted.


Assuntos
Encéfalo/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , S-Nitrosoglutationa/farmacologia , Acidente Vascular Cerebral/tratamento farmacológico , Animais , Humanos
8.
Plant Sci ; 292: 110389, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32005394

RESUMO

Nitric oxide (NO) is an intracellular messenger that mediates stress responses. Several plant aldehyde dehydrogenase (ALDH) genes are expressed during abiotic stress conditions to reduce the level of cytotoxic aldehydes. We investigated a possible interference between NO and ALDHs, using the isoform ALDH3H1 of Arabidopsis thaliana as model. The physiological NO donor; S-nitrosoglutathione (GSNO), inhibits ALDH3H1 in a time- and concentration-dependent manner. Mutagenesis and ESI-MS/MS analyses show that all Cys residues of ALDH3H1 are targets of GSNO-mediated S-nitrosation. Chemical labelling indicates that the deactivation is due to the conversion of the catalytic thiol into a catalytically non-active nitrosothiol. GSNO has the same effect on the chloroplastic ALDH3I1, suggesting that susceptibility of the catalytic Cys to NO is a common feature of ALDHs. S-Nitrosation and enzymatic inhibition of ALDH were reverted by reducing agents. Our study proves that the function of ALDHs does not exclusively depend on transcriptional regulation, with stress-induced expression, but may be also susceptible to posttranslational regulation through S-nitrosation. We discuss the potential involvement of S-nitrosoglutathione reductase (GSNOR), binding specific cofactors and reducing partners in a protective system of ALDHs in vivo, which will be experimentally corroborated in our forthcoming study.


Assuntos
Aldeído Desidrogenase/genética , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico/metabolismo , S-Nitrosoglutationa/farmacologia , Aldeído Desidrogenase/metabolismo , Arabidopsis/química , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Nitrosação , Estresse Fisiológico
9.
Molecules ; 25(3)2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-32013200

RESUMO

The beneficial cardiovascular effects of garlic have been reported in numerous studies. The major bioactive properties of garlic are related to organic sulfides. This study aimed to investigate whether garlic juice works exclusively due to its sulfur compounds or rather via the formation of new products of the nitroso-sulfide signaling pathway. Changes in isometric tension were measured on the precontracted aortic rings of adult normotensive Wistar rats. We evaluated NO-donor (S-nitrosoglutathione, GSNO)-induced vasorelaxation and compare it with effects of hydrogen sulfide (H2S)/GSNO and garlic/GSNO. Incubation with garlic juice increased the maximal GSNO-induced relaxation and markedly changed the character of the relaxant response. Although incubation with an H2S donor enhanced the maximal vasorelaxant response of GSNO, neither the absolute nor the relative relaxation changed over time. The mixture of GSNO with an H2S donor evoked a response similar to GSNO-induced relaxation after incubation with garlic juice. This relaxation of the H2S and GSNO mixture was soluble guanylyl cyclase (sGC) dependent, partially reduced by HNO scavenger and it was adenosine triphosphate-sensitive potassium channels (KATP) independent. In this study, we demonstrate for the first time the suggestion that H2S itself is probably not the crucial bioactive compound of garlic juice but rather potentiates the production of new signaling molecules during the GSNO-H2S interaction.


Assuntos
Alho , Sulfeto de Hidrogênio/farmacologia , Extratos Vegetais/farmacologia , S-Nitrosoglutationa/farmacologia , Transdução de Sinais , Vasodilatadores/farmacologia , Animais , Aorta/efeitos dos fármacos , Sucos de Frutas e Vegetais , Alho/química , Técnicas In Vitro , Contração Isométrica/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Vasodilatadores/isolamento & purificação
10.
Acta Biomater ; 103: 259-271, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31846803

RESUMO

Multidrug resistance of cancer cells is one of the major obstacle for chemotherapeutic efficiency. Nitric oxide (NO) has raised the potential to overcome multidrug resistance (MDR) with low side effects. Herein, we report a reactive oxygen species (ROS) and glutathione (GSH) responsive nanoparticle for the delivery of NO prodrug such as S-nitrosoglutathione (GSNO), which was chemically conjugated to an amphiphilic block copolymer. The GSNO functionalized nanoparticles show high NO loading capacity, good stability and sustained NO release with specific GSH activated NO-releasing kinetics. Such GSNO functionalized nanoparticles delivered doxorubicin (DOX) in a ROS triggered manner and increased the intracellular accumulation of DOX. However, in normal healthy cells, showing physiological concentrations of ROS, these nanoparticles presented good biocompatibility. The present work indicated that these multifunctional nanoparticles can serve as effective co-delivery platforms of NO and DOX to selectively kill chemo-resistant cancer cells through increasing chemo-sensitivity. STATEMENT OF SIGNIFICANCE: In this work, we constructed nitric oxide donor (S-nitrosoglutathione, GSNO) functionalized amphiphilic copolymer (PEG-PPS-GSNO) to deliver doxorubicin (DOX). The developed PEG-PPS-GSNO@DOX nanoparticles presented high NO capacity, ROS triggered DOX release and GSH triggered NO release. Thus NO reversed the chemo-resistance in HepG2/ADR cells increasing intrcellular accumulation of DOX. Furthermore, these PEG-PPS-GSNO@DOX nanoparticles exhibited biocompatibility to healthy cells and toxicity to cancer cells, due to elevated ROS.


Assuntos
Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Nanopartículas/química , Neoplasias/tratamento farmacológico , Polímeros/química , Espécies Reativas de Oxigênio/metabolismo , S-Nitrosoglutationa/farmacologia , Morte Celular/efeitos dos fármacos , Doxorrubicina/farmacologia , Liberação Controlada de Fármacos , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Endocitose/efeitos dos fármacos , Células HEK293 , Células Hep G2 , Humanos , Concentração Inibidora 50 , Nanopartículas/ultraestrutura , Neoplasias/patologia , Óxido Nítrico , Tamanho da Partícula , Polietilenoglicóis/síntese química , Polietilenoglicóis/química , Polímeros/síntese química , Espectroscopia de Prótons por Ressonância Magnética , Sulfetos/síntese química , Sulfetos/química
11.
J Stroke Cerebrovasc Dis ; 28(12): 104470, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31680031

RESUMO

BACKGROUND: The nitric oxide (NO)-producing activity of endothelial nitric oxide synthase (eNOS) plays a significant role in maintaining endothelial function and protecting against the stroke injury. However, the activity of the eNOS enzyme and the metabolism of major NO metabolite S-nitrosoglutathione (GSNO) are dysregulated after stroke, causing endothelial dysfunction. We investigated whether an administration of exogenous of GSNO or enhancing the level of endogenous GSNO protects against neurovascular injury in wild-type (WT) and eNOS-null (endothelial dysfunction) mouse models of cerebral ischemia-reperfusion (IR). METHODS: Transient cerebral ischemic injury was induced by middle cerebral artery occlusion (MCAO) for 60 minutes in male adult WT and eNOS null mice. GSNO (0.1 mg/kg body weight, intravenously) or N6022 (GSNO reductase inhibitor, 5.0 mg/kg body weight, intravenously) was administered 30 minutes before MCAO in preinjury and at the reperfusion in postinjury studies. Brain infarctions, edema, and neurobehavioral functions were evaluated at 24 hours after the reperfusion. RESULTS: eNOS-null mice had a higher degree (P< .05) of injury than WT. Pre- or postinjury treatment with either GSNO or N6022 significantly reduced infarct volume, improved neurological and sensorimotor function in both WT and eNOS-null mice. CONCLUSION: Reduced brain infarctions and edema, and improved neurobehavioral functions by pre- or postinjury GSNO treatment of eNOS knock out mice indicate that GSNO can attenuate IR injury, likely by mimicking the eNOS-derived NO-dependent anti-ischemic and anti-inflammatory functions. Neurovascular protection by GSNO/N6022 in both pre- and postischemic injury groups support GSNO as a promising drug candidate for the prevention and treatment of stroke injury.


Assuntos
Álcool Desidrogenase/antagonistas & inibidores , Benzamidas/farmacologia , Encéfalo/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Infarto da Artéria Cerebral Média/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Óxido Nítrico Sintase Tipo III/metabolismo , Óxido Nítrico/metabolismo , Pirróis/farmacologia , S-Nitrosoglutationa/farmacologia , Álcool Desidrogenase/metabolismo , Animais , Comportamento Animal/efeitos dos fármacos , Encéfalo/embriologia , Encéfalo/patologia , Edema Encefálico/enzimologia , Edema Encefálico/patologia , Edema Encefálico/prevenção & controle , Modelos Animais de Doenças , Infarto da Artéria Cerebral Média/enzimologia , Infarto da Artéria Cerebral Média/genética , Infarto da Artéria Cerebral Média/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico Sintase Tipo III/deficiência , Óxido Nítrico Sintase Tipo III/genética
12.
Int J Mol Sci ; 20(21)2019 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-31661878

RESUMO

Nitric oxide (NO) acts an essential signaling molecule that is involved in regulating various physiological and biochemical processes in plants. However, whether S-nitrosylation is a crucial molecular mechanism of NO is still largely unknown. In this study, 50 µM S-nitrosoglutathione (GSNO) treatment was found to have a maximum biological effect on promoting adventitious rooting in cucumber. Meanwhile, removal of endogenous NO significantly inhibited the development of adventitious roots implying that NO is responsible for promoting the process of adventitious rooting. Moreover, application of GSNO resulted in an increase of intracellular S-nitrosothiol (SNO) levels and endogenous NO production, while decreasing the S-nitrosoglutathione reductase (GSNOR) activity during adventitious rooting, implicating that S-nitrosylation might be involved in NO-induced adventitious rooting in cucumber. Furthermore, the identification of S-nitrosylated proteins was performed utilizing the liquid chromatography/mass spectrometry/mass spectrometry (LC-MS/MS) and biotin-switch technique during the development of adventitious rooting. Among these proteins, the activities and S-nitrosylated level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), tubulin alpha chain (TUA), and glutathione reductase (GR) were further analyzed as NO direct targets. Our results indicated that NO might enhance the S-nitrosylation level of GAPDH and GR, and was found to subsequently reduce these activities and transcriptional levels. Conversely, S-nitrosylation of TUA increased the expression level of TUA. The results implied that S-nitrosylation of key proteins seems to regulate various pathways through differential S-nitrosylation during adventitious rooting. Collectively, these results suggest that S-nitrosylation could be involved in NO-induced adventitious rooting, and they also provide fundamental evidence for the molecular mechanism of NO signaling during adventitious rooting in cucumber explants.


Assuntos
Cucumis sativus/metabolismo , Óxido Nítrico/metabolismo , Raízes de Plantas/efeitos dos fármacos , S-Nitrosoglutationa/farmacologia , Aldeído Oxirredutases/metabolismo , Cromatografia Líquida , Cucumis sativus/química , Cucumis sativus/efeitos dos fármacos , Cucumis sativus/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Proteoma/efeitos dos fármacos , Proteômica , S-Nitrosoglutationa/química , Transdução de Sinais , Espectrometria de Massas em Tandem , Tubulina (Proteína)/metabolismo
13.
Sci Rep ; 9(1): 15403, 2019 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-31659183

RESUMO

Murine 3T3-L1 adipocytes share many similarities with primary fat cells and represent a reliable in vitro model of adipogenesis. The aim of this study was to probe the effect of S-nitrosoglutathione (GSNO) on adipocyte differentiation. Adipogenesis was induced with a mixture of insulin, dexamethasone, and 3-isobutyl-1-methylxanthine in the absence and presence of increasing GSNO concentrations. Biochemical analysis after 7 days of differentiation showed a prominent anti-adipogenic effect of GSNO which was evident as reduced cellular triglycerides and total protein content as well as decreased mRNA and protein expression of late transcription factors (e.g. peroxisome proliferator activated receptor γ) and markers of terminal differentiation (e.g. leptin). By contrast, the nitrosothiol did not affect mRNA and protein expression of CCAAT/enhancer-binding protein ß (C/EBPß), which represents a pivotal early transcription factor of the adipogenic cascade. Differentiation was also inhibited by the NO donor (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate. Biotin switch experiments showed significantly increased S-nitrosation of C/EBPß variants indicating that posttranslational S-nitrosative modification of this transcription factor accounts for the observed anti-adipogenic effect of NO. Our results suggest that S-nitrosation might represent an important physiological regulatory mechanism of fat cell maturation.


Assuntos
Adipócitos/efeitos dos fármacos , Adipogenia , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Doadores de Óxido Nítrico/farmacologia , S-Nitrosoglutationa/farmacologia , 1-Metil-3-Isobutilxantina/farmacologia , Células 3T3-L1 , Adipócitos/citologia , Adipócitos/metabolismo , Animais , Proteína beta Intensificadora de Ligação a CCAAT/genética , Dexametasona/farmacologia , Insulina/farmacologia , Leptina/genética , Leptina/metabolismo , Camundongos , PPAR gama/genética , PPAR gama/metabolismo
14.
Am J Respir Cell Mol Biol ; 61(6): 765-775, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31596601

RESUMO

S-nitrosothiols (SNOs) are endogenous signaling molecules that have numerous beneficial effects on the airway via cyclic guanosine monophosphate-dependent and -independent processes. Healthy human airways contain SNOs, but SNO levels are lower in the airways of patients with cystic fibrosis (CF). In this study, we examined the interaction between SNOs and the molecular cochaperone C-terminus Hsc70 interacting protein (CHIP), which is an E3 ubiquitin ligase that targets improperly folded CF transmembrane conductance regulator (CFTR) for subsequent degradation. Both CFBE41o- cells expressing either wild-type or F508del-CFTR and primary human bronchial epithelial cells express CHIP. Confocal microscopy and IP studies showed the cellular colocalization of CFTR and CHIP, and showed that S-nitrosoglutathione inhibits the CHIP-CFTR interaction. SNOs significantly reduced both the expression and activity of CHIP, leading to higher levels of both the mature and immature forms of F508del-CFTR. In fact, SNO inhibition of the function and expression of CHIP not only improved the maturation of CFTR but also increased CFTR's stability at the cell membrane. S-nitrosoglutathione-treated cells also had more S-nitrosylated CHIP and less ubiquitinated CFTR than cells that were not treated, suggesting that the S-nitrosylation of CHIP prevents the ubiquitination of CFTR by inhibiting CHIP's E3 ubiquitin ligase function. Furthermore, the exogenous SNOs S-nitrosoglutathione diethyl ester and S-nitro-N-acetylcysteine increased the expression of CFTR at the cell surface. After CHIP knockdown with siRNA duplexes specific for CHIP, F508del-CFTR expression increased at the cell surface. We conclude that SNOs effectively reduce CHIP-mediated degradation of CFTR, resulting in increased F508del-CFTR expression on airway epithelial cell surfaces. Together, these findings indicate that S-nitrosylation of CHIP is a novel mechanism of CFTR correction, and we anticipate that these insights will allow different SNOs to be optimized as agents for CF therapy.


Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Processamento de Proteína Pós-Traducional , S-Nitrosotióis/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Aprotinina/farmacologia , Células Cultivadas , Fibrose Cística/genética , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Humanos , Leupeptinas/farmacologia , Dobramento de Proteína , Estabilidade Proteica , Proteólise , Interferência de RNA , RNA Interferente Pequeno/farmacologia , S-Nitrosoglutationa/farmacologia , Ubiquitina-Proteína Ligases/antagonistas & inibidores , Ubiquitina-Proteína Ligases/genética , Ubiquitinação
15.
Eur Rev Med Pharmacol Sci ; 23(17): 7674-7683, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31539160

RESUMO

OBJECTIVE: The aim of this study is to investigate the relation between CaMKII S-nitrosylation and its activation, as well as the underlying mechanism, after global cerebral ischemia-reperfusion. MATERIALS AND METHODS: The rat model of cerebral ischemia-reperfusion was established by four-vessel occlusion of 15 min and reperfusion of different times. nNOS inhibitor 7-nitroindazole (7-NI), exogenous nitric oxide donor GSNO (nitrosoglutathione), or N-methyl-D-aspartate receptor (NMDAR) antagonist MK-801 were administered before ischemia. The expressions of S-nitrosylation and phosphorylation of CaMKII and nNOS were detected by biotin switch assay, immunoblotting, and immunohistochemical staining after cerebral ischemia-reperfusion. The survival of hippocampal CA1 pyramidal cells after administration of the three drugs was examined by cresyl violet staining. RESULTS: Following cerebral ischemia-reperfusion, the S-nitrosylation of CaMKII was increased, accompanied by a decrease of phosphorylation, suggesting a decrease of activity (p<0.05). Meanwhile, the phosphorylation and S-nitrosylation of nNOS were notably decreased at the same time point (p<0.05). The administration of 7-NI, GSNO, and MK-801 increased the S-nitrosylation and phosphorylation of nNOS, leading to the attenuation of increased S-nitrosylation and decreased autophosphorylation of CaMKII after cerebral ischemia-reperfusion (p<0.05). Administration of MK-801, GSNO, and 7-NI significantly decreased the neuronal damage in rat hippocampal CA1 caused by cerebral ischemia-reperfusion (p<0.05). CONCLUSIONS: After cerebral ischemia-reperfusion, the decrease of autophosphorylation of CaMKII regulated by its S-nitrosylation may be due to the denitrosylation of nNOS and subsequent NO production. Increasing the phosphorylation of CaMKII by nNOS inhibitor, exogenous NO donor or NMDA receptor antagonist exerted neuroprotective effects against cerebral ischemia-reperfusion injury.


Assuntos
Região CA1 Hipocampal/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Óxido Nítrico Sintase Tipo I/metabolismo , Traumatismo por Reperfusão/patologia , Animais , Região CA1 Hipocampal/patologia , Modelos Animais de Doenças , Maleato de Dizocilpina/farmacologia , Indazóis/farmacologia , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Fosforilação/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/metabolismo , S-Nitrosoglutationa/farmacologia
16.
Biomolecules ; 9(9)2019 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-31438648

RESUMO

S-nitrosoglutathione reductase (GSNOR) exerts crucial roles in the homeostasis of nitric oxide (NO) and reactive nitrogen species (RNS) in plant cells through indirect control of S-nitrosation, an important protein post-translational modification in signaling pathways of NO. Using cultivated and wild tomato species, we studied GSNOR function in interactions of key enzymes of reactive oxygen species (ROS) metabolism with RNS mediated by protein S-nitrosation during tomato root growth and responses to salinity and cadmium. Application of a GSNOR inhibitor N6022 increased both NO and S-nitrosothiol levels and stimulated root growth in both genotypes. Moreover, N6022 treatment, as well as S-nitrosoglutathione (GSNO) application, caused intensive S-nitrosation of important enzymes of ROS metabolism, NADPH oxidase (NADPHox) and ascorbate peroxidase (APX). Under abiotic stress, activities of APX and NADPHox were modulated by S-nitrosation. Increased production of H2O2 and subsequent oxidative stress were observed in wild Solanum habrochaites, together with increased GSNOR activity and reduced S-nitrosothiols. An opposite effect occurred in cultivated S. lycopersicum, where reduced GSNOR activity and intensive S-nitrosation resulted in reduced ROS levels by abiotic stress. These data suggest stress-triggered disruption of ROS homeostasis, mediated by modulation of RNS and S-nitrosation of NADPHox and APX, underlies tomato root growth inhibition by salinity and cadmium stress.


Assuntos
Aldeído Oxirredutases/metabolismo , Cádmio/toxicidade , Lycopersicon esculentum/efeitos dos fármacos , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Cloreto de Sódio/farmacologia , Ascorbato Peroxidases/metabolismo , Benzamidas/química , Benzamidas/metabolismo , Benzamidas/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Lycopersicon esculentum/crescimento & desenvolvimento , Lycopersicon esculentum/metabolismo , NADPH Oxidases/metabolismo , Óxido Nítrico/metabolismo , Nitrosação , Estresse Oxidativo/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Pirróis/química , Pirróis/metabolismo , Pirróis/farmacologia , Espécies Reativas de Nitrogênio/química , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/química , S-Nitrosoglutationa/farmacologia , S-Nitrosotióis/metabolismo , Solanum/crescimento & desenvolvimento , Solanum/metabolismo , Estresse Fisiológico
17.
Biochem Pharmacol ; 168: 352-365, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31374192

RESUMO

The epigenetic signatures associated with cognitive decline driven by lack of estrogen in post-menopausal state, is not well-understood. The present study is an attempt to unravel the epigenetic mechanisms involved in cognitive impairment preceding ovariectomy in mice and evaluate the protective effects of S-nitrosoglutathione (GSNO). A significant decline in cognitive functions was observed in mice following ovariectomy as assessed by Morris water maze and Novel object recognition test. Administration of GSNO (100 µg/kg body weight, orally) daily for 4 weeks was found to ameliorate cognitive deficits observed in ovariectomised (OVX) mice. The activity of histone acetyl-transferase (HAT) was significantly disrupted in cortex and hippocampus of OVX mice. This was accompanied by increased activity of histone deacetylase (HDAC) and increased levels of HDAC-2, HDAC-3 causing lowered acetylated histone (H)3 levels. Reduced H3 acetylation triggers epigenetic repression of brain derived neurotrophic factor (BDNF) in cortex and hippocampus of OVX mice that may be responsible for neuronal damage and cognitive impairment. GSNO supplementation to OVX mice was able to reinstate HAT(CBP/p300) and HDAC balance through S-nitrosylation. GSNO restored histone acetylation at BDNF promoters (pII, pIV) thereby ameliorating BDNF levels and improving brain morphology and cognition. The study suggests that GSNO improves cognitive function in OVX mice by modulating epigenetic programming.


Assuntos
Disfunção Cognitiva/prevenção & controle , Epigênese Genética/efeitos dos fármacos , S-Nitrosoglutationa/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Fator Neurotrófico Derivado do Encéfalo/sangue , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Imunoprecipitação da Cromatina , Estradiol/sangue , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Histona Desacetilases/genética , Histona Desacetilases/metabolismo , Histonas/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ovariectomia , S-Nitrosoglutationa/administração & dosagem
18.
Nitric Oxide ; 90: 1-9, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31170453

RESUMO

Currently, most antimicrobial topical treatments utilize antibiotics to prevent or treat infection at a wound site. However, with the ongoing evolution of multi-drug resistant bacterial strains, there is a high demand for alternative antimicrobial treatments. Nitric oxide (NO) is an endogenous gas molecule with potent antimicrobial activity, which is effective against a wide variety of bacterial strains. In this study, the potential for creating NO releasing creams containing the naturally occurring NO carrier, S-nitrosoglutathione (GSNO), are characterized and evaluated. GSNO is shown to have prolonged stability (>300 days) when mixed and stored within Vaseline at 24 °C. Further, enhanced proliferation of NO from GSNO using zinc oxide nanoparticles (ZnO) is demonstrated. Triggering NO release from the GSNO/Vaseline mixture using a commercial zinc oxide-containing cream exhibits first-order NO release kinetics with the highest %NO release over the first 6 h. Significant killing effects against S. aureus, S. epidermidis, and P. aeruginosa are demonstrated for the GSNO/Vaseline/ZnO cream mixtures in a proportional manner dependent upon the concentration of GSNO in the final mixture.


Assuntos
Antibacterianos/farmacologia , Óxido Nítrico/metabolismo , S-Nitrosoglutationa/farmacologia , Óxido de Zinco/farmacologia , Antibacterianos/química , Antibacterianos/metabolismo , Testes de Sensibilidade Microbiana , Óxido Nítrico/química , Pseudomonas aeruginosa/efeitos dos fármacos , S-Nitrosoglutationa/química , S-Nitrosoglutationa/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Óxido de Zinco/química , Óxido de Zinco/metabolismo
19.
J Pharm Pharmacol ; 71(8): 1255-1261, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31115903

RESUMO

OBJECTIVES: To investigate the therapeutic effects and mechanisms of S-nitrosoglutathione (SNG) on acute kidney injury (AKI) induced by lipopolysaccharide (LPS). METHODS: We established an AKI model by intraperitoneal administration of LPS in mice and LPS-induced human kidney (HK-2) cells in vitro. We obtained the kidney tissues from mice for histopathological examination, examined inflammatory cytokines by enzyme-linked immunosorbent assay and measured the expression levels of toll-like receptor 4-nuclear factor-κB (TLR4-NF-κB) signal pathway-related proteins by Western blotting. KEY FINDINGS: Pretreatment of SNG effectively improved the kidney function, reduced the pathological damage score of kidney in mice and decreased the expression levels of IL-1ß, IL-6 and TNF-α in a dose-dependent manner in vivo and in vitro. Furthermore, pretreatment of SNG also repressed TLR4, phosphorylated NF-κB IκBα, IKKß and p65 expression levels in HK-2 cells induced by LPS. CONCLUSIONS: S-nitrosoglutathione attenuates the severity of LPS-induced AKI by inhibiting the TLR4-NF-κB signalling pathway and may act as a protective agent for septic AKI.


Assuntos
Injúria Renal Aguda/tratamento farmacológico , NF-kappa B/metabolismo , Substâncias Protetoras/farmacologia , S-Nitrosoglutationa/farmacologia , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/metabolismo , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/metabolismo , Animais , Linhagem Celular , Citocinas/metabolismo , Humanos , Proteínas I-kappa B/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fator de Necrose Tumoral alfa/metabolismo
20.
Cell Death Dis ; 10(5): 329, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30988280

RESUMO

Protein S-nitrosylation, the redox-based posttranslational modification of a cysteine thiol by the attachment of a nitric oxide (NO) group, is responsible for a variety of signaling effects. Dysregulation of S-nitrosylation may be directly linked to cancer apoptotic resistance and cancer therapy outcomes, emphasizing the importance of S-nitrosylation in cancer. Peroxiredoxin-2 (Prdx2), an antioxidant enzyme, plays an important role in the protection of cancer cells from oxidative radical damage caused by hydrogen dioxide (H2O2), which is a potential target for cancer therapy. Our studies showed that, as an endogenous NO carrier, S-nitrosoglutathione (GSNO) induced apoptosis in lung cancer cells via nitrosylating Prdx2. The nitrosylation of Prdx2 at Cys51 and Cys172 sites disrupted the formation of Prdx2 dimer and repressed the Prdx2 antioxidant activity, causing the accumulation of endogenous H2O2. H2O2 activated AMPK, which then phosphorylated SIRT1 and inhibited its deacetylation activity toward p53 in A549 cells or FOXO1 in NCI-H1299 cells. Taken together, our results elucidate the roles and mechanisms of Prdx2 S-nitrosylation at Cys51 and Cys172 sites in lung cancer cells apoptosis and this finding provides an effective lung cancer treatment strategy for managing aberrant Prdx2 activity in lung cancers.


Assuntos
Apoptose/efeitos dos fármacos , Peroxirredoxinas/metabolismo , S-Nitrosoglutationa/farmacologia , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/antagonistas & inibidores , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular , Proteína Forkhead Box O1/metabolismo , Humanos , Peróxido de Hidrogênio/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Óxido Nítrico/metabolismo , Peroxirredoxinas/antagonistas & inibidores , Peroxirredoxinas/genética , Fosforilação , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Sirtuína 1/antagonistas & inibidores , Sirtuína 1/genética , Sirtuína 1/metabolismo
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