RESUMO
Here, we report for the first time on the mechanisms of action of the essential oil of Ruta graveolens (REO) against the plant pathogen Colletotrichum gloeosporioides. In particular, the presence of REO drastically affected the morphology of hyphae by inducing changes in the cytoplasmic membrane, such as depolarization and changes in the fatty acid profile where straight-chain fatty acids (SCFAs) increased by up to 92.1%. In addition, REO induced changes in fungal metabolism and triggered apoptosis-like responses to cell death, such as DNA fragmentation and the accumulation of reactive oxygen species (ROS). The production of essential enzymes involved in fungal metabolism, such as acid phosphatase, ß-galactosidase, ß-glucosidase, and N-acetyl-ß-glucosaminidase, was significantly reduced in the presence of REO. In addition, C. gloeosporioides activated naphthol-As-BI phosphohydrolase as a mechanism of response to REO stress. The data obtained here have shown that the essential oil of Ruta graveolens has a strong antifungal effect on C. gloeosporioides. Therefore, it has the potential to be used as a surface disinfectant and as a viable replacement for fungicides commonly used to treat anthracnose in the postharvest testing phase.
Assuntos
Antifúngicos , Colletotrichum , Óleos Voláteis , Espécies Reativas de Oxigênio , Ruta , Colletotrichum/efeitos dos fármacos , Óleos Voláteis/farmacologia , Óleos Voláteis/química , Ruta/química , Antifúngicos/farmacologia , Antifúngicos/química , Espécies Reativas de Oxigênio/metabolismo , Doenças das Plantas/microbiologia , Testes de Sensibilidade Microbiana , Fragmentação do DNA/efeitos dos fármacosRESUMO
Experiments conducted on triple-negative breast cancer have shown that fucoidan from Lessonia trabeculata (FLt) exhibits cytotoxic and antitumor properties. However, further research is necessary to gain a complete understanding of its bioactivity and level of cytotoxicity. The cytotoxic effect of FLt was determined by the 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Apoptosis was analyzed using annexin V and caspase 3/7 staining kit and DNA fragmentation. In addition, transcriptional expression of antiapoptotic (Bcl-2 and XIAP) and proapoptotic (caspase 8, caspase 9, and AIF) genes were analyzed in TNBC 4T1 cells. After 72 h of culture, the IC50 for FLt was 561 µg/mL, while doxorubicin (Dox) had an IC50 of 0.04 µg/mL. In addition, assays for FLt + Dox were performed. Annexin V and caspase 3/7 revealed that FLt induces early and late-stage apoptosis. DNA fragmentation results support necrotic death of 4T1 cells. Similarly, transcripts that prevent cell death were decreased, while transcripts that promote cell death were increased. This study showed that FLt induces apoptosis by both caspase-dependent and caspase-independent mechanisms. These findings suggest that FLt may have potential applications in breast cancer treatment. Further research will provide more information to elucidate the mechanism of action of FLt.
Assuntos
Apoptose , Caspases , Polissacarídeos , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Polissacarídeos/farmacologia , Animais , Feminino , Caspases/metabolismo , Camundongos , Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Humanos , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/patologia , Fragmentação do DNA/efeitos dos fármacos , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/patologia , KelpRESUMO
Short time treatment with reduced dosages of selol-loaded PLGA nanocapsules (NcSel) combined with magnetic hyperthermia (MHT) is evaluated in aged Erhlich tumor-bearing mice. Clinical, hematological, biochemical, genotoxic and histopathological parameters are assessed during 7 d treatment with NcSel and MHT, separately or combined. The time evolution of the tumor volume is successfully modeled using the logistic mathematical model. The combined therapy comprising NcSel and MHT is able to hinder primary tumor growth and a case of complete tumor remission is recorded. Moreover, no metastasis was diagnosed and the adverse effects are negligible. NcSel plus MHT may represent an effective and safe alternative to cancer control in aged patients. Future clinical trials are encouraged.
Assuntos
Neoplasias da Mama/terapia , Hipertermia Induzida , Nanopartículas de Magnetita/uso terapêutico , Nanocápsulas/uso terapêutico , Compostos de Selênio/uso terapêutico , Animais , Neoplasias da Mama/patologia , Carcinoma de Ehrlich/patologia , Carcinoma de Ehrlich/terapia , Ciclo Celular/efeitos dos fármacos , Terapia Combinada , Fragmentação do DNA/efeitos dos fármacos , Feminino , Nanopartículas de Magnetita/química , Nanopartículas de Magnetita/ultraestrutura , Camundongos , Nanocápsulas/química , Nanocápsulas/ultraestrutura , Compostos de Selênio/química , Fatores de Tempo , Resultado do Tratamento , Carga Tumoral/efeitos dos fármacosRESUMO
Quinoline and 1,2,3-triazoles are well-known nitrogen-based heterocycles presenting diverse pharmacological properties, although their antileishmanial activity is still poorly exploited. As an effort to contribute with studies involving these interesting chemical groups, in the present study, a series of compounds derived from 4-aminoquinoline and 1,2,3-triazole were synthetized and biological studies using L. amazonensis species were performed. The results pointed that the derivative 4, a hybrid of 4-aminoquinoline/1,2,3-triazole exhibited the best antileishmanial action, with inhibitory concentration (IC50) values of ~1 µM against intramacrophage amastigotes of L. amazonensis , and being 16-fold more active to parasites than to the host cell. The mechanism of action of derivative 4 suggest a multi-target action on Leishmania parasites, since the treatment of L. amazonensis promastigotes caused mitochondrial membrane depolarization, accumulation of ROS products, plasma membrane permeabilization, increase in neutral lipids, exposure of phosphatidylserine to the cell surface, changes in the cell cycle and DNA fragmentation. The results suggest that the antileishmanial effect of this compound is primarily altering critical biochemical processes for the correct functioning of organelles and macromolecules of parasites, with consequent cell death by processes related to apoptosis-like and necrosis. No up-regulation of reactive oxygen and nitrogen intermediates was promoted by derivative 4 on L. amazonensis -infected macrophages, suggesting a mechanism of action independent from the activation of the host cell. In conclusion, data suggest that derivative 4 presents selective antileishmanial effect, which is associated with multi-target action, and can be considered for future studies for the treatment against disease.
Assuntos
Aminoquinolinas/farmacologia , Antiprotozoários/farmacologia , Leishmania mexicana/efeitos dos fármacos , Triazóis/farmacologia , Aminoquinolinas/síntese química , Animais , Antiprotozoários/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Membrana Celular/química , Membrana Celular/metabolismo , Fragmentação do DNA/efeitos dos fármacos , Feminino , Metabolismo dos Lipídeos/efeitos dos fármacos , Macrófagos/parasitologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Organelas/efeitos dos fármacos , Fosfatidilserinas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Triazóis/síntese químicaRESUMO
Quinones are plant-derived secondary metabolites that present diverse pharmacological properties, including antibacterial, antifungal, antiviral, anti-inflammatory, antipyretic and anticancer activities. In the present study, we evaluated the cytotoxic effect of a new naphthoquinone 6b,7-dihydro-5H-cyclopenta [b]naphtho [2,1-d]furan-5,6 (9aH)-dione) (CNFD) in different tumor cell lines. CNFD displayed cytotoxic activity against different tumor cell lines, especially in MCF-7 human breast adenocarcinoma cells, which showed IC50 values of 3.06 and 0.98 µM for 24 and 48 h incubation, respectively. In wound-healing migration assays, CNFD promoted inhibition of cell migration. We have found typical hallmarks of apoptosis, such as cell shrinkage, chromatin condensation, phosphatidylserine exposure, increase of caspases-9 and-3 activation, increase of internucleosomal DNA fragmentation without affecting the cell membrane permeabilization, increase of ROS production, and loss of mitochondrial membrane potential induced by CNFD. Moreover, gene expression experiments indicated that CNFD increased the expression of the genes CDKN1A, FOS, MAX, and RAC1 and decreased the levels of mRNA transcripts of several genes, including CCND1, CDK2, SOS1, RHOA, GRB2, EGFR and KRAS. The CNFD treatment of MCF-7 cells induced the phosphorylation of c-jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinases (MAPKs) and inactivation of extracellular signal-regulated protein kinase 1/2 (ERK1/2). In a study using melanoma cells in a murine model in vivo, CNFD induced a potent anti-tumor activity. Herein, we describe, for the first time, the cytotoxicity and anti-tumor activity of CNFD and sequential mechanisms of apoptosis in MCF-7 cells. CNFD seems to be a promising candidate for anti-tumor therapy.
Assuntos
Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Melanoma/tratamento farmacológico , Naftoquinonas/uso terapêutico , Espécies Reativas de Oxigênio/metabolismo , Animais , Antineoplásicos/farmacologia , Caspases/metabolismo , Linhagem Celular Tumoral , DNA/metabolismo , Fragmentação do DNA/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , MAP Quinase Quinase 4/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Mitocôndrias/metabolismo , Naftoquinonas/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
BACKGROUND: In the coming decades, diabetes mellitus might affect 628 million individuals. Its final impact on male fertility and reproductive outcomes should be considered since the number of adolescents and young adults presenting diabetes is rising. Resveratrol (RES), a polyphenol, is a biological modulator with multitarget and multi-action characteristics. OBJECTIVES: to evaluate if RES is effective against the male reproductive damage caused by type 1 diabetes (DM1), focusing on sperm DNA integrity and reproductive outcome. MATERIALS AND METHODS: At 30 dpp (days postpartum), male rats were divided into 7 groups: Sham control (SC); RES vehicle (RV); RES (R); STZ-diabetic (D; induced at 30dpp with 65 mg/kg of streptozotocin); STZ-diabetic + insulin (DI); STZ-diabetic + RES (DR); STZ-diabetic + insulin +RES (DIR). DR, DIR, and R groups received 150mg RES/kg b.w./day by gavage (from 33 to 110dpp). DI and DIR received insulin (from day 5 after DM1 induction until 110dpp). Blood glucose was monitored in different time points. Animals were mated with healthy females. Euthanasia occurred at 110 dpp. RESULTS: DM1 increased lipid peroxidation (testis and epididymis) and sperm DNA fragmentation, alterations of chromatin structure, reduced mitochondrial mass and acrosome integrity, causing a decline in fertility and pregnancy rates. RES improved the parameters. DISCUSSION: RES, as an adjuvant, activates specific reactions against hyperglycemia, the main trigger of most complications of diabetes, by controlling oxidative stress, probably as a result of SIRT1 activation. We present here more evidences showing its valuable role in diminishing diabetes seriousness to male reproduction, not only to spermatogenesis in the first instance, but also to sperm overall quality and fertility outcomes, regardless of insulin treatment. CONCLUSION: RES attenuated lipid peroxidation and sperm DNA damage in DM1-induced animals, which positively reflected on male fertility. Our results show RES potential against DM1 complications in male reproduction.
Assuntos
Antioxidantes/uso terapêutico , Diabetes Mellitus Tipo 1/complicações , Infertilidade Masculina/tratamento farmacológico , Resveratrol/uso terapêutico , Espermatozoides/efeitos dos fármacos , Animais , Antioxidantes/farmacologia , Fragmentação do DNA/efeitos dos fármacos , Diabetes Mellitus Experimental , Avaliação Pré-Clínica de Medicamentos , Infertilidade Masculina/etiologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Ratos Wistar , Reprodução/efeitos dos fármacos , Resveratrol/farmacologiaRESUMO
BACKGROUND AND OBJECTIVE: Evidence point out promising anticancer activities of Dihydropyrimidinones (DHPM) and organoselenium compounds. This study aimed to evaluate the cytotoxic and antiproliferative potential of DHPM-derived selenoesters (Se-DHPM), as well as their molecular mechanisms of action. METHODS: Se-DHPM cytotoxicity was evaluated against cancer lines (HeLa, HepG2, and MCF-7) and normal cells (McCoy). HepG2 clonogenic assay allowed verifying antiproliferative effects. The propidium iodide/ orange acridine fluorescence readings showed the type of cell death induced after treatments (72h). Molecular simulations with B-DNA and 49H showed docked positions (AutoDock Vina) and trajectories/energies (GROMACS). In vitro molecular interactions used CT-DNA and 49H applying UV-Vis absorbance and fluorescence. Comet assay evaluated DNA fragmentation of HepG2 cells. Flow cytometry analysis verified HepG2 cell cycle effects. Levels of proteins (ß-actin, p53, BAX, HIF-1α, γH2AX, PARP-1, cyclin A, CDK-2, and pRB) were quantified by immunoblotting. RESULTS: Among Se-DHPM, 49H was selectively cytotoxic to HepG2 cells, reduced cell proliferation, and increased BAX (80%), and p53 (66%) causing apoptosis. Molecular assays revealed 49H inserted in the CT-DNA molecule causing the hypochromic effect. Docking simulations showed H-bonds and hydrophobic interactions, which kept the ligand partially inserted into the DNA minor groove. 49H increased the DNA damage (1.5 fold) and γH2AX level (153%). Besides, treatments reduced PARP-1 (60%) and reduced pRB phosphorylation (21%) as well as decreased cyclin A (46%) arresting cell cycle at the G1 phase. CONCLUSION: Together all data obtained confirmed the hypothesis of disruptive interactions between Se-DHPM and DNA, thereby highlighting its potential as a new anticancer drug.
Assuntos
Antineoplásicos/síntese química , Carcinoma Hepatocelular/tratamento farmacológico , Citotoxinas/síntese química , Neoplasias Hepáticas/tratamento farmacológico , Compostos Organosselênicos/síntese química , Actinina/metabolismo , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Citotoxinas/farmacologia , Fragmentação do DNA/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Simulação de Acoplamento Molecular , Compostos Organosselênicos/farmacologia , Compostos Organosselênicos/toxicidade , Fosforilação , Relação Estrutura-Atividade , Proteína X Associada a bcl-2/metabolismoRESUMO
B-N-methylamino-L-alanine (BMAA), a cyanotoxin produced by most cyanobacteria, has been proposed to cause long term damages leading to neurodegenerative diseases, including Amyotrophic Lateral Sclerosis/Parkinsonism Dementia complex (ALS/PDC) and retinal pathologies. Previous work has shown diverse mechanisms leading to BMAA-induced degeneration; however, the underlying mechanisms of toxicity affecting retina cells are not fully elucidated. We here show that BMAA treatment of rat retina neurons in vitro induced nuclear fragmentation and cell death in both photoreceptors (PHRs) and amacrine neurons, provoking mitochondrial membrane depolarization. Pretreatment with the N-Methyl-D-aspartate (NMDA) receptor antagonist MK-801 prevented BMAA-induced death of amacrine neurons, but not that of PHRs, implying activation of NMDA receptors participated only in amacrine cell death. Noteworthy, BMAA stimulated a selective axonal outgrowth in amacrine neurons, simultaneously promoting growth cone destabilization. BMAA partially decreased the viability of Müller glial cells (MGC), the main glial cell type in the retina, induced marked alterations in their actin cytoskeleton and impaired their capacity to protect retinal neurons. BMAA also induced cell death and promoted axonal outgrowth in differentiated rat pheochromocytoma (PC12) cells, implying these effects were not limited to amacrine neurons. These results suggest that BMAA is toxic for retina neurons and MGC and point to the involvement of NMDA receptors in amacrine cell death, providing new insight into the mechanisms involved in BMAA neurotoxic effects in the retina.
Assuntos
Diamino Aminoácidos/toxicidade , Células Ependimogliais/efeitos dos fármacos , Agonistas de Aminoácidos Excitatórios/toxicidade , Doenças Retinianas/induzido quimicamente , Neurônios Retinianos/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Sobrevivência Celular/efeitos dos fármacos , Toxinas de Cianobactérias , Fragmentação do DNA/efeitos dos fármacos , Maleato de Dizocilpina/farmacologia , Células Ependimogliais/patologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Doenças Retinianas/metabolismo , Doenças Retinianas/prevenção & controle , Neurônios Retinianos/patologiaRESUMO
To verify a possible synergistic effect of smoking and varicocele on the seminal plasma proteome and biological functions, a cross-sectional study was performed in 25 smokers and 24 nonsmokers. Samples were used for conventional semen analysis, functional analysis (DNA fragmentation, acrosome integrity and mitochondrial activity) and proteomics by a shotgun approach. Functional enrichment of biological pathways was performed in differentially expressed proteins. Smokers presented lower ejaculate volume (p = .027), percentage of progressively motile spermatozoa (p = .002), total sperm count (p = .039), morphology (p = .001) and higher percentage of immotile spermatozoa (p = .03), round cell (p = .045) and neutrophil count (p = .009). Smokers also presented lower mitochondrial activity and acrosome integrity and higher DNA fragmentation. We identified and quantified 421 proteins in seminal plasma, of which one was exclusive, 21 were overexpressed and 70 were underexpressed in the seminal plasma of smokers. The proteins neprilysin, beta-defensin 106A and histone H4A were capable of predicting the smoker group. Enriched functions were related to immune function and sperm machinery in testis/epididymis. Based on our findings, we can conclude that cigarette smoking leads to the establishment of inflammatory protein pathways in the testis/epididymis in the presence of varicocele that seems to act in synergy with the toxic components of the cigarette.
Assuntos
Fumar Cigarros/efeitos adversos , Infertilidade Masculina/imunologia , Sêmen/química , Proteínas de Plasma Seminal/análise , Varicocele/complicações , Acrossomo/efeitos dos fármacos , Acrossomo/imunologia , Acrossomo/patologia , Adulto , Brasil , Estudos Transversais , Fragmentação do DNA/efeitos dos fármacos , Epididimo/irrigação sanguínea , Epididimo/efeitos dos fármacos , Epididimo/imunologia , Humanos , Infertilidade Masculina/patologia , Masculino , Pessoa de Meia-Idade , não Fumantes/estatística & dados numéricos , Proteômica/estatística & dados numéricos , Sêmen/imunologia , Sêmen/metabolismo , Análise do Sêmen/estatística & dados numéricos , Proteínas de Plasma Seminal/metabolismo , Transdução de Sinais/imunologia , Fumantes/estatística & dados numéricos , Testículo/irrigação sanguínea , Testículo/efeitos dos fármacos , Testículo/imunologia , Nicotiana/toxicidade , Varicocele/imunologia , Adulto JovemRESUMO
(-)-Epicatechin is a phenolic compound with antioxidant activity that is present in natural food and drinks, such as cocoa and red wine. Evidence suggests that (-)-epicatechin exhibits anticancer activity; however, its mechanism of action is poorly understood. Here, we investigated the anticancer effects of (-)-epicatechin and its mechanism of action in breast cancer cells. We assessed the anticancer activity by cell proliferation assays, apoptosis by DNA fragmentation and flow cytometry. The expression of proteins associated with apoptosis was analyzed by the human apoptosis array. MitoSOXTM Red and biomarkers of oxidative damage were used to measure the effect of (-)-epicatechin on mitochondrial reactive oxygen species (ROS) and cellular damage, respectively. (-)-Epicatechin treatment caused a decreasing in the viability of MDA-MB-231 and MCF-7 cells. This cell death was associated with DNA fragmentation and an apoptotic proteomic profile. Further, (-)-epicatechin in MDA-MB-231 cells upregulated death receptor (DR4/DR5), increased the ROS production, and modulated pro-apoptotic proteins. In MCF-7 cells, (-)-epicatechin did not involve death receptor; however, an increase in ROS and the upregulation of pro-apoptotic proteins (Bad and Bax) were observed. These changes were associated with the apoptosis activation through the intrinsic pathway. In conclusion, this study shows that (-)-epicatechin has anticancer activity in breast cancer cells and provides novel insight into the molecular mechanism of (-)-epicatechin to induce apoptosis.