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1.
Life Sci ; 269: 119080, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33465387

RESUMO

S-Allylcysteine (SAC) is an extensively studied natural product which has been proven to confer cardioprotection. This potentiates SAC into many clinical relevance possibilities, hence, the use of it ought to be optimally elucidated. To further confirm this, an ischemia/reperfusion model has been used to determine SAC at 10 mM and 50 mM on cardiac function, cardiac marker, and mitochondrial permeability. Using Langendorff setup, 24 adult male Wistar rats' hearts were isolated to be perfused with Kreb-Henseleit buffer throughout the ischemia/reperfusion method. After 20 min of stabilization, global ischemia was induced by turning off the perfusion for 35 min followed by 60 min of reperfusion with either Kreb-Henseleit buffer or SAC with the dose of 10 mM or 50 mM. The cardiac function was assessed and coronary effluent was collected at different timepoints throughout the experiment for lactate dehydrogenase (LDH) measurement. The harvested hearts were then used to measure glutathione while isolated mitochondria for mPTP analysis. SAC-reperfused hearts were shown to prevent the aggravation of cardiac function after I/R induction. It also dose-dependently upregulated glutathione reductase and glutathione level and these were also accompanied by significant reduction of LDH leakage and preserved mitochondrial permeability. Altogether, SAC dose-dependently was able to recover the post-ischemic cardiac function deterioration alongside with improvement of glutathione metabolism and mitochondrial preservation. These findings highly suggest that SAC when sufficiently supplied to the heart would be able to prevent the deleterious complications after the ischemic insult.


Assuntos
Antioxidantes/farmacologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Cisteína/análogos & derivados , Coração/efeitos dos fármacos , Isquemia/tratamento farmacológico , Mitocôndrias/efeitos dos fármacos , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Animais , Antineoplásicos/farmacologia , Cisteína/farmacologia , Coração/fisiopatologia , Isquemia/metabolismo , Isquemia/patologia , Masculino , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Ratos , Ratos Wistar
2.
Food Chem ; 338: 128055, 2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-32950008

RESUMO

This study examined the ability of l-arginine, l-cysteine and l-methionine, to inhibit postharvest senescence of broccoli. Florets were dipped in aqueous solutions of the amino acids at concentrations from 1.0 to 100 mM and stored at 10 °C. A 5 mM dip was found to be optimal in delaying senescence as measured by retention of green colour, vitamin C and antioxidant activity, and a lower level of ethylene production, respiration, weight loss, phenylalanine ammonia lyase (PAL) activity and ion leakage with the benefits being similar for all three amino acids. Arginine, cysteine and methionine have Generally Recognised As Safe (GRAS) status and should have few impediments in obtaining regulatory approval for commercial use if similar effects were found for other leafy vegetables.


Assuntos
Arginina/farmacologia , Brassica/efeitos dos fármacos , Cisteína/farmacologia , Metionina/farmacologia , Proteínas de Plantas/metabolismo , Amônia-Liases/metabolismo , Antioxidantes/química , Antioxidantes/metabolismo , Ácido Ascórbico/metabolismo , Brassica/metabolismo , Etilenos/metabolismo , Fatores de Tempo
3.
Ecotoxicol Environ Saf ; 209: 111784, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33316727

RESUMO

Cysteine (Cys) is incorporated into several compounds which are involved in detoxification of heavy metals. It is evident from recent studies that Cys is effective in alleviating the toxicity of heavy metals. Nevertheless, little is known about the Cys-mediated alleviation of chromium (Cr) toxicity. In our study, the impacts of exogenous Cys on Cr-stressed maize (Zea mays L.) were examined by using physiological and proteomic analyses. The results showed that Cr (100 µM) increased the accumulation of hydrogen peroxide, decreased cell viability, enhanced lipid peroxidation and consequently inhibited plant growth. The application of Cys (500 µM) attenuated the adverse effects of Cr on seedling growth. Cys supplementation to Cr treated plants decreased Cr accumulation in the shoots and increased Cr accumulation in roots. Cys treatment also modulated the activities of antioxidant enzymes and increased endogenous Cys content. Sixty proteins in root tissue were significantly affected by exogenous Cys under Cr stress using two-dimensional electrophoresis. Forty-six differentially expressed proteins were successfully identified by MALDI-TOF/TOF mass spectrometry. These differentially expressed proteins were involved in various biological pathways such as stress response (41.3%), energy and carbohydrate metabolism (21.7%), protein metabolism (6.5%), amino acid metabolism (6.5%), and others of unknown functions. The defense response-related proteins including glutathione peroxidase, glutathione S-transferases, pathogenesis-related proteins, glyoxalases and superoxide dismutase were differently regulated by Cys suggesting their roles in the Cys-mediated Cr tolerance.


Assuntos
Antioxidantes/farmacologia , Cromo/toxicidade , Cisteína/farmacologia , Poluentes do Solo/toxicidade , Zea mays/fisiologia , Antioxidantes/metabolismo , Cromo/metabolismo , Cisteína/metabolismo , Glutationa Peroxidase/metabolismo , Peróxido de Hidrogênio/metabolismo , Lactoilglutationa Liase/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Proteoma/efeitos dos fármacos , Proteômica , Plântula/efeitos dos fármacos , Superóxido Dismutase/metabolismo , Zea mays/metabolismo
4.
Sheng Li Xue Bao ; 72(3): 299-307, 2020 Jun 25.
Artigo em Chinês | MEDLINE | ID: mdl-32572428

RESUMO

The purpose of the present study is to investigate the effect of L-cysteine on colonic motility and the underlying mechanism. Immunohistochemical staining and Western blot were used to detect the localization of the H2S-generating enzymes cystathionine-ß-synthase (CBS) and cystathionine-γ-lyase (CSE). Organ bath system was used to observe the muscle contractile activities. Whole-cell patch-clamp technique was applied to record ionic channels currents in colonic smooth muscle cells. The results showed that both CBS and CSE were localized in mucosa, longitudinal and circular muscle and enteric neurons. L-cysteine had a dual effect on colonic contraction, and the excitatory effect was blocked by pretreatment with CBS inhibitor aminooxyacetate acid (AOAA) and CSE inhibitor propargylglycine (PAG); L-cysteine concentration-dependently inhibited L-type calcium channel current (ICa,L) without changing the characteristic of L-type calcium channel (P < 0.01); In contrast, the exogenous H2S donor NaHS increased ICa,L at concentration of 100 µmol/L, but inhibited ICa,L and modified the channel characteristics at concentration of 300 µmol/L (P < 0.05); Furthermore, L-cysteine had no effect on large conductance calcium channel current (IBKCa), but NaHS significantly inhibited IBKCa (P < 0.05). These results suggest that L-cysteine has a potential dual effect on colonic smooth muscle and the inhibitory effect might be directly mediated by L-type calcium channel while the excitatory effect might be mediated by endogenous H2S.


Assuntos
Cisteína/farmacologia , Sulfeto de Hidrogênio , Cistationina beta-Sintase , Cistationina gama-Liase , Músculo Liso
5.
Biotechniques ; 69(2): 108-112, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32459144

RESUMO

The outbreak of viral pneumonia caused by the novel coronavirus SARS-CoV-2 that began in December 2019 caused high mortality. It has been suggested that the main protease (Mpro) of SARS-CoV-2 may be an important target to discover pharmaceutical compounds for the therapy of this life-threatening disease. Remdesivir, ritonavir and chloroquine have all been reported to play a role in suppressing SARS-CoV-2. Here, we applied a molecular docking method to study the binding stability of these drugs with SARS-CoV-2 Mpro. It appeared that the ligand-protein binding stability of the alliin and SARS-CoV-2 Mpro complex was better than others. The results suggested that alliin may serve as a good candidate as an inhibitor of SARS-CoV-2 Mpro. Therefore, the present research may provide some meaningful guidance for the prevention and treatment of SARS-CoV-2.


Assuntos
Antivirais/farmacologia , Cisteína/análogos & derivados , Proteínas não Estruturais Virais/antagonistas & inibidores , Monofosfato de Adenosina/análogos & derivados , Monofosfato de Adenosina/farmacologia , Alanina/análogos & derivados , Alanina/farmacologia , Antimaláricos/farmacologia , Betacoronavirus/enzimologia , Cloroquina/farmacologia , Cisteína/farmacologia , Cisteína Endopeptidases , Simulação de Acoplamento Molecular , Ritonavir/farmacologia
6.
Food Chem Toxicol ; 137: 111180, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32032653

RESUMO

Aflatoxins are carcinogenic metabolites produced by Aspergillus and Penicillium spp. Aflatoxin contamination of food is a serious health hazard. Some metal ions (such as Zn2+) affect Aspergillus growth and aflatoxin biosynthesis. Presence of zinc in the growth medium incites aflatoxin production. This study investigates the effect of zinc binding amino acids and peptides on aflatoxin synthesis in indigenous toxigenic Aspergillus species isolated from agro-ecological zones in Northern Iran. Zinc (II) chelating nutrients (such as Histidine (His), Cysteine (Cys), Histidine-Cysteine (His-Cys), and triple peptide (Asn-Cys-Ser) were added to the growth medium of toxigenic Aspergillus isolates and incubated at temperature range of 25-40 °C. Aflatoxin production on different culture media was tested using ELISA. Addition of cysteine to Sabouraud dextrose broth (SDB) medium significantly reduced aflatoxin production, which could be related to its zinc chelating property. Aflatoxin production was drastically restrained at high concentration of His, especially in combination with Cys, at high pH values and incubation temperature (pH = 7.5, temperature = 40 °C). Aflatoxin production was low in presence of triple peptide (Asn-Cys-Ser) at concentration of 500 mg/L. From the application perspective, natural zinc chelators can be used as harmless aflatoxin-production inhibitors.


Assuntos
Aflatoxina B1/antagonistas & inibidores , Aspergillus flavus/efeitos dos fármacos , Quelantes/farmacologia , Cisteína/farmacologia , Histidina/farmacologia , Oligopeptídeos/farmacologia , Aflatoxina B1/metabolismo , Quelantes/química , Zinco/química
7.
Int J Mol Sci ; 21(3)2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-32041330

RESUMO

The S-Allyl-L-cysteine ​​(SAC) component of aged garlic extract (AGE) is proven to have anticancer, antihepatotoxic, neuroprotective and neurotrophic properties. -Cystathionase (CTH), cystathionine ß-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase (MPST) are involved in H2S/sulfane sulfur endogenous formation from L-cysteine. The aim of the study was to determine the effect of SAC on MCF-7 cells survival and apoptosis, which is a widely known approach to reduce the number of cancer cells. An additional goal of this paper was to investigate the effect of SAC on the activity and expression of enzymes involved in H2S production. The experiments were carried out in the human breast adenocarcinoma cell line MCF-7. Changes in the cell viability were determined by MTT assay. Cell survival was determined by flow cytometry (FC). Changes in enzymes expression were analyzed using Western blot. After 24 h and 48 h incubation with 2245 µM SAC, induction of late apoptosis was observed. A decrease in cell viability was observed with increasing SAC concentration and incubation time. SAC had no significant cytotoxic effect on the MCF-7 cells upon all analyzed concentrations. CTH, MPST and CBS expression were confirmed in non-treated MCF-7 cells. Significant decrease in MPST activity at 2245 µM SAC after 24 h and 48 h incubation vs. 1000 µM SAC was associated with decrease in sulfane sulfur levels. The presented results show promising SAC effects regarding the deterioration of the MCF-7 cells' condition in reducing their viability through the downregulation of MPST expression and sulfate sulfur level reduction.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias da Mama/tratamento farmacológico , Sobrevivência Celular/efeitos dos fármacos , Cisteína/análogos & derivados , Sulfurtransferases/biossíntese , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cisteína/farmacologia , Humanos , Sulfeto de Hidrogênio/metabolismo , Células MCF-7 , Extratos Vegetais/farmacologia
8.
Life Sci ; 245: 117367, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32001265

RESUMO

AIMS: The present study determines the effect of administration of novel antioxidant astaxanthin-s-allyl cysteine biconjugate (AST-SAC) against streptozotocin-induced diabetes mellitus (DM) in rats. MAIN METHODS: AST-SAC (1 mg/kg/day) was treated against DM in rats for 45 days. The oxidative stress, antioxidants level, insulin secretion, activities of various carbohydrate metabolizing enzymes were studied. The glucose uptake in L6 myotubes was studied. In addition, in silico analysis of interaction of AST-SAC with proteins such as insulin receptor (IR) and 5'-adenosine monophosphate-activated protein kinase (AMPK) were carried out. KEY FINDINGS: Administration of AST-SAC in DM rats has protected the mitochondrial function (decreased oxidative stress and normalized oxidative phosphorylation activities) and antioxidant capacity of the pancreas which has resulted in beta cells rejuvenation and insulin secretion restoration. AST-SAC decreased the alpha-glucosidases activities to bring glycemic control in DM rats. Due to these effects the glycoprotein components and lipids were restored to near normalcy in DM rats. AST-SAC protected the antioxidant status of liver, kidney and plasma; and curbed the progression of secondary complications of DM. AST-SAC treatment stimulated glucose uptake in L6 myotubes in in vitro. To support this observation, AST-SAC interacted with proteins such as IR and AMPK in silico. SIGNIFICANCE: AST-SAC can be considered as "multi-target-directed ligand", that is, through these manifold effects, AST-SAC has been able to prevail over DM in rats.


Assuntos
Antioxidantes/uso terapêutico , Cisteína/análogos & derivados , Cisteína/uso terapêutico , Diabetes Mellitus Experimental/tratamento farmacológico , Hipoglicemiantes/uso terapêutico , Mitocôndrias/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Xantofilas/uso terapêutico , Animais , Antioxidantes/farmacologia , Colesterol/metabolismo , Cisteína/farmacologia , Diabetes Mellitus Experimental/metabolismo , Glucose/metabolismo , Masculino , Mitocôndrias/metabolismo , Simulação de Acoplamento Molecular , Ratos , Ratos Sprague-Dawley , Triglicerídeos/metabolismo , Xantofilas/farmacologia
9.
Eur J Pharmacol ; 869: 172893, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31883915

RESUMO

Organosulfur compounds, such as L-cysteine, allicin and other sulfur-containing organic compounds in Allium species, have been proposed to possess many important physiological and pharmacological functions. A novel L-cysteine derivative, t-Butyl S-allylthio-L-cysteinate (5P39), was designed and synthesized by combining L-cysteine derivative and allicin pharmacophore through a disulfide bond. This study aimed to explore the effects and mechanisms of 5P39 on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. At the experimental concentration (5, 10 and 20 µM), 5P39 suppressed the excessive secretion of nitric oxide (NO) and interleukin-6 (IL-6) in mice peritoneal macrophages stimulated by LPS. A mouse model of ALI was established by tracheal instillation of LPS for 2 h before 5P39 (30 and 60 mg/kg) administration. The results showed that 5P39 treatment down-regulated the wet/dry weight ratio (W/D ratio) of lungs and reduced the protein concentration, the number of total cells as well as the myeloperoxidase (MPO) activity in bronchoalveolar lavage fluid (BALF). 5P39 administration improved the histopathological changes of lungs in ALI mice with the decreased levels of pro-inflammatory cytokines in BALF. The inhibitory effects of 5P39 on the toll-like receptor 4 (TLR4) expression and macrophages accumulation in lung tissues were observed by immunohistochemistry. Additionally, 5P39 significantly attenuated the LPS-activated high expression of key proteins in TLR4/MyD88 signaling pathway. Taken together, the present study showed that 5P39 effectively alleviate the severity of ALI, and its mechanism might relate to the inhibition of LPS-activated TLR4/MyD88 signaling pathway, demonstrating a promising potential for further development into an anti-inflammatory drug candidate.


Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Cisteína/farmacologia , Cisteína/uso terapêutico , Fator 88 de Diferenciação Mieloide/metabolismo , Receptor 4 Toll-Like/metabolismo , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/patologia , Animais , Líquido da Lavagem Broncoalveolar/química , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cisteína/análogos & derivados , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/metabolismo , Masculino , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
10.
Biochem Biophys Res Commun ; 521(2): 414-419, 2020 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-31672277

RESUMO

Lysosome-associated membrane protein-2 (LAMP2), is a highly glycosylated lysosomal membrane protein involved in chaperone mediated autophagy. Mutations of LAMP2 cause the classic triad of myopathy, cardiomyopathy and encephalopathy of Danon disease (DD). Additionally, retinopathy has also been observed in young DD patients, leading to vision loss. Emerging evidence show LAMP2-deficiency to be involved in oxidative stress (ROS) but the mechanism remains obscure. In the present study, we found that tert-butyl hydroperoxide or antimycin A induced more cell death in LAMP2 knockdown (LAMP2-KD) than in control ARPE-19 cells. Mechanistically, LAMP2-KD reduced the concentration of cytosolic cysteine, resulting in low glutathione (GSH), inferior antioxidant capability and mitochondrial lipid peroxidation. ROS induced RPE cell death through ferroptosis. Inhibition of glutathione peroxidase 4 (GPx4) increased lethality in LAMP2-KD cells compared to controls. Cysteine and glutamine supplementation restored GSH and prevented ROS-induced cell death of LAMP2-KD RPE cells.


Assuntos
Ferroptose , Proteína 2 de Membrana Associada ao Lisossomo/genética , Espécies Reativas de Oxigênio/efeitos adversos , Epitélio Pigmentado da Retina/patologia , Linhagem Celular , Cisteína/farmacologia , Técnicas de Silenciamento de Genes , Glutamina/farmacologia , Glutationa/metabolismo , Humanos , Epitélio Pigmentado da Retina/citologia
11.
Spectrochim Acta A Mol Biomol Spectrosc ; 225: 117516, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31518754

RESUMO

We describe a sensitive turn-on fluorescent assay for antioxidants by using fluorescence-tunable graphene quantum dots (GQDs). GQDs exhibited strong fluorescence without dopamine (DA). DA could self-polymerize to a thin polydopamine (PDA) film on the surface of GQDs under alkaline environment, resulting in the fluorescence quenching of GQDs via fluorescence resonance energy transfer (FRET). However, the self-polymerization of DA could be effectively inhibited in the presence of antioxidants including glutathione (GSH), ascorbic acid (AA), cysteine (Cys), and homocysteine (Hcys). Thus, the fluorescence of GQDs restored. The "turn-on" sensing of antioxidants could be achieved with high sensitivity. The detection limit for GSH, AA, Cys, and Hcys could be achieved as low as 2.4 nM, 1.5 nM, 4.2 nM, and 4.4 nM, respectively. Finally, the GQDs@PDA system was applied for monitoring cerebral antioxidants in rat brain microdialysates. This work promises new opportunities to evaluate antioxidant capacity in physiological and pathological fields.


Assuntos
Antioxidantes/análise , Dopamina/química , Pontos Quânticos/química , Antioxidantes/farmacologia , Ácido Ascórbico/análise , Ácido Ascórbico/farmacologia , Química Encefálica , Cisteína/análise , Cisteína/farmacologia , Estudos de Viabilidade , Fluorescência , Transferência Ressonante de Energia de Fluorescência , Corantes Fluorescentes/química , Glutationa/análise , Glutationa/farmacologia , Grafite , Homocisteína/análise , Homocisteína/farmacologia , Indóis/química , Limite de Detecção , Microdiálise , Polimerização/efeitos dos fármacos , Polímeros/química , Pontos Quânticos/ultraestrutura
12.
J Med Chem ; 63(5): 2282-2291, 2020 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-31418565

RESUMO

Cancer immunotherapy has gained increasing attention due to its potential specificity and lack of adverse side effects when compared to more traditional modes of treatment. Toll-like receptor 2 (TLR2) agonists are lipopeptides possessing the S-[2,3-bis(palmitoyloxy)propyl]-l-cysteine (Pam2Cys) motif and exhibit potent immunostimulatory effects. These agonists offer a means of providing "danger signals" in order to activate the immune system toward tumor antigens. Thus, the development of TLR2 agonists is attractive in the search of potential immunostimulants for cancer. Existing SAR studies of Pam2Cys with TLR2 indicate that the structural requirements for activity are, for the most part, very intolerable. We have investigated the importance of stereochemistry, the effect of N-terminal acylation, and homologation between the two ester functionalities in Pam2Cys-conjugated lipopeptides on TLR2 activity. The R diastereomer is significantly more potent than the S diastereomer and N-terminal modification generally lowers TLR2 activity. Most notably, homologation gives rise to analogues which are comparatively active to the native Pam2Cys containing constructs.


Assuntos
Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/farmacologia , Lipopeptídeos/química , Lipopeptídeos/farmacologia , Receptor 2 Toll-Like/agonistas , Adjuvantes Imunológicos/síntese química , Vacinas Anticâncer/farmacologia , Cisteína/análogos & derivados , Cisteína/síntese química , Cisteína/farmacologia , Humanos , Lipopeptídeos/síntese química , Neoplasias/prevenção & controle , Estereoisomerismo , Receptor 2 Toll-Like/metabolismo
13.
Adv Clin Exp Med ; 28(12): 1609-1614, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31851788

RESUMO

BACKGROUND: During cerebral ischemia, energy restoration through the regulation of glucose transporters and antioxidant defense mechanisms is essential to maintain cell viability. Antioxidant therapy has been considered effective to attenuate brain damage; moreover, the regulation of transcription factors that positively regulate the expression of glucose transporters is associated with this therapy. Recently, it has been reported that the use of antioxidants such as S-allylcysteine (SAC), a component of aged garlic extract (AGE), improves survival in experimental models of cerebral ischemia. OBJECTIVES: The aim of this study was to determine the effect of AGE and SAC on the level of mRNA expression of the main neuronal glucose transporter (GLUT3) and the glutamate cysteine ligase catalytic subunit (GCLC) in rats with transient focal cerebral ischemia. MATERIAL AND METHODS: Cerebral ischemia was induced in male Wistar rats by middle cerebral artery occlusion (MCAO) for 2 h. The animals were sacrificed after different reperfusion times (0-48 h). Animals injected with AGE (360 mg/kg, intraperitoneally (i.p.)) and SAC (300 mg/kg, i.p.) at the beginning of reperfusion were sacrificed after 2 h. The mRNA expression level was analyzed in the fronto-parietal cortex using quantitative polymerase chain reaction (qPCR). RESULTS: Two major increases in GLUT3 expression at 1 h and 24 h of reperfusion were found. Both treatments increased GLUT3 and GCLC mRNA levels in control and under ischemic/reperfusion injury animals. CONCLUSIONS: This data suggests that SAC and AGE might induce neuroprotection, while controlling reactive oxygen species (ROS) levels, as indicated by the increase in GCLC expression, and regulating the energy content of the cell by increasing glucose transport mediated by GLUT3.


Assuntos
Isquemia Encefálica , Alho , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Glutamato-Cisteína Ligase/metabolismo , Fármacos Neuroprotetores , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Isquemia Encefálica/metabolismo , Cisteína/análogos & derivados , Cisteína/farmacologia , Alho/química , Proteínas Facilitadoras de Transporte de Glucose/efeitos dos fármacos , Glutamato-Cisteína Ligase/efeitos dos fármacos , Masculino , Fármacos Neuroprotetores/uso terapêutico , Extratos Vegetais/farmacologia , Ratos , Ratos Wistar , Traumatismo por Reperfusão/metabolismo
14.
J Mol Med (Berl) ; 97(12): 1643-1656, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31773180

RESUMO

Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic renal disease, caused in the majority of the cases by a mutation in either the PKD1 or the PKD2 gene. ADPKD is characterised by a progressive increase in the number and size of cysts, together with fibrosis and distortion of the renal architecture, over the years. This is accompanied by alterations in a complex network of signalling pathways. However, the underlying molecular mechanisms are not well characterised. Previously, we defined the PKD Signature, a set of genes typically dysregulated in PKD across different disease models from a meta-analysis of expression profiles. Given the importance of transcription factors (TFs) in modulating disease, we focused in this paper on characterising TFs from the PKD Signature. Our results revealed that out of the 1515 genes in the PKD Signature, 92 were TFs with altered expression in PKD, and 32 of those were also implicated in tissue injury/repair mechanisms. Validating the dysregulation of these TFs by qPCR in independent PKD and injury models largely confirmed these findings. STAT3 and RUNX1 displayed the strongest activation in cystic kidneys, as demonstrated by chromatin immunoprecipitation (ChIP) followed by qPCR. Using immunohistochemistry, we showed a dramatic increase of expression after renal injury in mice and cystic renal tissue of mice and humans. Our results suggest a role for STAT3 and RUNX1 and their downstream targets in the aetiology of ADPKD and indicate that the meta-analysis approach is a viable strategy for new target discovery in PKD. KEY MESSAGES: We identified a list of transcription factors (TFs) commonly dysregulated in ADPKD. Out of the 92 TFs identified in the PKD Signature, 35% are also involved in injury/repair processes. STAT3 and RUNX1 are the most significantly dysregulated TFs after injury and during PKD progression. STAT3 and RUNX1 activity is increased in cystic compared to non-cystic mouse kidneys. Increased expression of STAT3 and RUNX1 is observed in the nuclei of renal epithelial cells, also in human ADPKD samples.


Assuntos
Subunidade alfa 2 de Fator de Ligação ao Core/metabolismo , Regulação da Expressão Gênica/genética , Rim/metabolismo , Rim Policístico Autossômico Dominante/metabolismo , Fator de Transcrição STAT3/metabolismo , Fatores de Transcrição/metabolismo , Animais , Imunoprecipitação da Cromatina , Subunidade alfa 2 de Fator de Ligação ao Core/genética , Cisteína/análogos & derivados , Cisteína/farmacologia , Cisteína/toxicidade , Modelos Animais de Doenças , Progressão da Doença , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Humanos , Rim/efeitos dos fármacos , Rim/lesões , Masculino , Camundongos , Camundongos Transgênicos , Rim Policístico Autossômico Dominante/genética , Regiões Promotoras Genéticas/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/genética , Fator de Transcrição STAT3/genética , Canais de Cátion TRPP/genética , Fatores de Transcrição/genética
15.
Int J Mol Med ; 44(5): 1943-1951, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31573046

RESUMO

S­allyl­cysteine sulfoxide (alliin) is the main organosulfur component of garlic and its preparations. The present study aimed to examine the protective effect of alliin on cardiac function and the underlying mechanism in a mouse model of myocardial infarction (MI). Notably, alliin treatment preserved heart function, attenuated the area of infarction in the myocardium of mice and reduced lesions in the myocardium, including cardiomyocyte fibrosis and death. Further mechanistic experiments revealed that alliin inhibited necroptosis but promoted autophagy in vitro and in vivo. Cell viability assays showed that alliin dose­dependently reduced the necroptotic index and inhibited the expression of necroptosis­related receptor­interacting protein 1, receptor­interacting protein 3 and tumor necrosis factor receptor­associated factor 2, whereas the levels of Beclin 1 and microtubule­associated protein 1 light chain 3, which are associated with autophagy, exhibited an opposite trend upon treatment with alliin. In addition, the level of peroxisome proliferator­activated receptor γ was increased by alliin. Collectively, these findings demonstrate that alliin has the potential to protect cardiomyocytes from necroptosis following MI and that this protective effect occurs via the enhancement of autophagy.


Assuntos
Autofagia/efeitos dos fármacos , Cisteína/análogos & derivados , Infarto do Miocárdio/tratamento farmacológico , Miócitos Cardíacos/efeitos dos fármacos , Necroptose/efeitos dos fármacos , Safrol/análogos & derivados , Animais , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cisteína/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Safrol/farmacologia , Transdução de Sinais/efeitos dos fármacos
16.
Eur J Pharmacol ; 863: 172707, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31568786

RESUMO

Hydrogen sulfide plays an important role in the regulation of the cardiovascular system, insulin secretion, and glucose homeostasis. The aim of the present study was to examine the effects of chronic treatment with sodium hydrosulfide (NaHS), L-Cysteine (L-Cys) and DL-Propargylglycine (DL-PAG) on the changes induced by a high-fat diet (HFD) in zoometric and metabolic variables as well as cardiovascular changes such as hypertension and sympathetic hyperactivity. For this purpose, male Wistar rats were fed a normal fat diet (NFD) or HFD for 12 weeks. Next, the HFD rats were divided into 5 subgroups which received daily i.p. injections during 4 weeks of: (1) nothing (no injection, Control); (2) vehicle (PBS; 1ml/kg); (3) NaHS (5.6 mg/kg); (4) L-Cys (300mg/kg); or (5) DL-PAG (1mg/kg). Then, an oral glucose tolerance test, hormone serum levels and blood pressure were determined. The cardiovascular responses to stimulation of the vasopressor sympathetic tone or intravenous administration of the agonists noradrenaline (α1/2-adrenoceptors), methoxamine (α1-adrenoceptors) and UK 14,304 (α2-adrenoceptors) were determined in pithed rats. Lastly, the heart, liver and adipose tissue were weighted. HFD significantly increased: (1) zoometric variables, which were decreased by NaHS and L-Cys; (2) metabolic variables, ameliorated by DL-PAG; (3) haemodynamic variables, which were reversed by NaHS and L-Cys; and (4) the vasopressor responses induced by sympathetic stimulation, which were diminished by NaHS and L-Cys. In conclusion, chronic treatment with NaHS and L-Cys are effective in reducing adipose tissue and ameliorating the cardiovascular changes induced by obesity; meanwhile, DL-PAG ameliorates metabolic variables.


Assuntos
Sistema Cardiovascular/efeitos dos fármacos , Cisteína/administração & dosagem , Cisteína/farmacologia , Dieta Hiperlipídica/efeitos adversos , Recuperação de Função Fisiológica/efeitos dos fármacos , Sulfetos/administração & dosagem , Sulfetos/farmacologia , Agonistas de Receptores Adrenérgicos alfa 1/farmacologia , Agonistas de Receptores Adrenérgicos alfa 2/farmacologia , Animais , Sistema Cardiovascular/fisiopatologia , Hemodinâmica/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Fatores de Tempo
17.
Int J Med Sci ; 16(8): 1180-1187, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31523181

RESUMO

Objective: The effects of pre-treatments from s-methyl cysteine (SMC) alone, syringic acid (SA) alone and SMC plus SA against kainic acid (KA) induced injury in nerve growth factor (NGF) differentiated PC12 cells were investigated. Methods: NGF-differentiated PC12 cells were treated with 1 µM SMC, 1 µM SA or 0.5 µM SMC plus 0.5 µM SA for 2 days. Subsequently, cells were further treated by 150 µM KA. Results: KA suppressed Bcl-2 mRNA expression, enhanced Bax mRNA expression and casued cell death. SMC was greater than SA, and similar as SMC+SA in increasing Bcl-2 mRNA expression. SMC+SA led to greater increase in mitochondrial membrane potential and cell survival than SMC or SA alone. SMC+SA resulted in more reduction in reactive oxygen species and tumor necrosis factor-alpha generation, more increase in glutathione content and glutathione reductase activity than SMC or SA alone. KA up-regulated protein expression of nuclear factor kappa B (NF-κB) p65 and phosphorylated p38 (p-p38). SMC or SA pre-treatments alone limited protein expression of both factors. SMC+SA resulted in more suppression in NF-κB p65 and p-p38 expression. KA decreased glutamine level, increased glutamate level and stimulated calcium release. SMC pre-treatments alone reversed these alterations. SMC alone elevated glutamine synthetase (GS) activity and mRNA expression. SMC+SA led to greater GS activity and mRNA expression than SMC pre-treatments alone. Conclusion: These findings suggested that this combination, SMC+SA, might provide greater protective potent for neuronal cells.


Assuntos
Cisteína/análogos & derivados , Ácido Gálico/análogos & derivados , Fator de Crescimento Neural/farmacologia , Fármacos Neuroprotetores/farmacologia , Animais , Cálcio/metabolismo , Diferenciação Celular/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/patologia , Sobrevivência Celular , Cisteína/farmacologia , Sinergismo Farmacológico , Ácido Gálico/farmacologia , Ácido Caínico/toxicidade , Potencial da Membrana Mitocondrial/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Células PC12 , Ratos , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
18.
PLoS One ; 14(9): e0221831, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31490969

RESUMO

The preservation of biological samples for an extended time period of days to weeks after initial collection is important for the identification, screening, and characterization of bacterial pathogens. Traditionally, preservation relies on cold-chain infrastructure; however, in many situations this is impractical or not possible. Thus, our goal was to develop alternative bacterial sample preservation and transport media that are effective without refrigeration or external instrumentation. The viability, nucleic acid stability, and protein stability of Bacillus anthracis Sterne 34F2, Francisella novicida U112, Staphylococcus aureus ATCC 43300, and Yersinia pestis KIM D27 (pgm-) was assessed for up to 28 days. Xanthan gum (XG) prepared in PBS with L-cysteine maintained more viable F. novicida U112 cells at elevated temperature (40°C) compared to commercial reagents and buffers. Viability was maintained for all four bacteria in XG with 0.9 mM L-cysteine across a temperature range of 22-40°C. Interestingly, increasing the concentration to 9 mM L-cysteine resulted in the rapid death of S. aureus. This could be advantageous when collecting samples in the built environment where there is the potential for Staphylococcus collection and stabilization rather than other organisms of interest. F. novicida and S. aureus DNA were stable for up to 45 days upon storage at 22°C or 40°C, and direct analysis by real-time qPCR, without DNA extraction, was possible in the XG formulations. XG was not compatible with proteomic analysis via LC-MS/MS due to the high amount of residual Xanthomonas campestris proteins present in XG. Our results demonstrate that polysaccharide-based formulations, specifically XG with L-cysteine, maintain bacterial viability and nucleic acid integrity for an array of both Gram-negative and Gram-positive bacteria across ambient and elevated temperatures.


Assuntos
Bactérias/efeitos dos fármacos , Polissacarídeos/farmacologia , Preservação Biológica/métodos , Bactérias/citologia , Bactérias/metabolismo , Cisteína/farmacologia , Viabilidade Microbiana/efeitos dos fármacos , Polissacarídeos Bacterianos/farmacologia , Proteômica , Temperatura
19.
Biochemistry ; 58(35): 3669-3682, 2019 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-31386812

RESUMO

Cancer cells regulate key enzymes in the glycolytic pathway to control the glycolytic flux, which is necessary for their growth and proliferation. One of the enzymes is pyruvate kinase muscle isoform 2 (PKM2), which is allosterically regulated by various small molecules. Using detailed biochemical and kinetic studies, we demonstrate that cysteine inhibits wild-type (wt) PKM2 by shifting from an active tetramer to a mixture of a tetramer and a less active dimer/monomer equilibrium and that the inhibition is dependent on cysteine concentration. The cysteine-mediated PKM2 inhibition is reversed by fructose 1,6-bisphosphate, an allosteric activator of PKM2. Furthermore, kinetic studies using two dimeric PKM2 variants, S437Y PKM2 and G415R PKM2, show that the reversal is caused by the tetramerization of wtPKM2. The crystal structure of the wtPKM2-Cys complex was determined at 2.25 Å, which showed that cysteine is held to the amino acid binding site via its main chain groups, similar to that observed for phenylalanine, alanine, serine, and tryptophan. Notably, ligand binding studies using fluorescence and isothermal titration calorimetry show that the presence of phosphoenolpyruvate alters the binding affinities of amino acids for wtPKM2 and vice versa, thereby unravelling the existence of a functionally bidirectional coupling between the amino acid binding site and the active site of wtPKM2.


Assuntos
Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/química , Cisteína/farmacologia , Inibidores Enzimáticos/farmacologia , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/química , Hormônios Tireóideos/química , Substituição de Aminoácidos/genética , Aminoácidos/química , Aminoácidos/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Catálise , Domínio Catalítico/genética , Cristalografia por Raios X , Humanos , Isoenzimas/antagonistas & inibidores , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Ligantes , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Modelos Moleculares , Proteínas Mutantes/antagonistas & inibidores , Proteínas Mutantes/química , Proteínas Mutantes/metabolismo , Fosfoenolpiruvato/química , Fosfoenolpiruvato/metabolismo , Ligação Proteica , Conformação Proteica , Serina/genética , Hormônios Tireóideos/genética , Hormônios Tireóideos/metabolismo , Tirosina/genética
20.
Toxins (Basel) ; 11(9)2019 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-31461909

RESUMO

Two garlic-derived compounds, Propyl Propane Thiosulfonate (PTS) and Propyl Propane Thiosulfinate (PTSO), were examined for their efficacy against mycotoxigenic Fusarium species (F. graminearum, F. langsethiae, F. verticillioides). The objectives were to assess the inhibitory effect of these compounds on growth and mycotoxin production in vitro, and in situ in artificially inoculated wheat, oats and maize with one isolate of each respectively, at different water activity (aw) conditions when stored for up to 20 days at 25 °C. In vitro, 200 ppm of either PTS or PTSO reduced fungal growth by 50-100% and mycotoxin production by >90% depending on species, mycotoxin and aw conditions on milled wheat, oats and maize respectively. PTS was generally more effective than PTSO. Deoxynivalenol (DON) and zearalenone (ZEN) were decreased by 50% with 80 ppm PTSO. One-hundred ppm of PTS reduced DON and ZEN production in wheat stored at 0.93 aw for 20 days, although contamination was still above the legislative limits. Contrasting effects on T-2/HT-2 toxin contamination of oats was found depending on aw, with PTS stimulating production under marginal conditions (0.93 aw), but at 0.95 aw effective control was achieved with 100 ppm. Treatment of stored maize inoculated with F. verticilliodies resulted in a stimulation of total fumonsins in most treatments. The potential use of such compounds for mycotoxin control in stored commodities is discussed.


Assuntos
Cisteína/análogos & derivados , Grão Comestível/microbiologia , Contaminação de Alimentos/análise , Fusarium/efeitos dos fármacos , Alho/química , Micotoxinas/análise , Ácidos Sulfínicos/farmacologia , Avena/microbiologia , Cisteína/isolamento & purificação , Cisteína/farmacologia , Fusarium/crescimento & desenvolvimento , Fusarium/metabolismo , Ácidos Sulfínicos/isolamento & purificação , Toxina T-2/análogos & derivados , Toxina T-2/análise , Triticum/microbiologia , Zea mays/microbiologia
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