RESUMO
Bisphenol A (BPA) may adversely affect human health by inducing oxidative stress and irreversible damage to cells. Bioactive compounds found in some functional foods, individually or in combination, can attenuate the negative effects of BPA exposure; an example is the multi-supplement containing guarana (Gua), selenium (Se), and L-carnitine (LC) -GSC- which has already demonstrated antioxidant, genoprotective, and immunomodulatory activities. This study aimed to determine the effect of GSC and its constituents on oxidative and genotoxic alterations triggered by BPA exposure in the retinal epithelial cell line. The cells exposed to BPA (0.001, 0.01, 0.1, 1, 3, and 10 µM) to determine the lowest concentration required to induce cyto-genotoxicity. ARPE-19 cells were then concomitantly exposed to the selected BPA concentration, GSC, and its components (Gua, 1.07 mg/mL; Se, 0.178 µg/mL; and LC, 1.43 mg/mL). Flow cytometry, biochemical assays, qRT-PCR, genotoxicity, apoptosis, and cellular proliferation. Based on our results, 10 µM of BPA could induce cyto-genotoxic and oxidative alterations. BPA did not alter the Bcl-2/BAX expression ratio but induced Casp3 and Casp8 overexpression, suggesting that apoptosis was induced mainly via the extrinsic pathway. GSC partially reversed the alterations triggered by BPA in ARPE-19 cells. However, Se had unexpected negative effects on ARPE-19 cells. The multi-supplement GSC may attenuate changes in oxidative and genotoxic markers related to exposure of ARPE-19 cells to BPA. our results revealed that the antioxidant, anti-apoptotic, and genoprotective properties of GSC were not universally shared by its individual, once Se did not exhibit any positive impact.
Assuntos
Apoptose , Compostos Benzidrílicos , Carnitina , Estresse Oxidativo , Fenóis , Epitélio Pigmentado da Retina , Selênio , Fenóis/toxicidade , Compostos Benzidrílicos/toxicidade , Humanos , Selênio/farmacologia , Carnitina/farmacologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Linhagem Celular , Paullinia/química , Dano ao DNA/efeitos dos fármacos , Antioxidantes/farmacologia , Células Epiteliais/efeitos dos fármacos , Citometria de Fluxo , Suplementos NutricionaisRESUMO
Guarana (Paullinia cupana) is habitually ingested by people in the Amazon region and is a key ingredient in various energy drinks consumed worldwide. Extension in longevity and low prevalence of chronic age-related diseases have been associated to habitual intake of guarana. Anti-aging potential of guarana was also demonstrated in Caenorhabditis elegans; however, the mechanisms involved in its effects are not clear. Herein, we investigated the putative pathways that regulate the effects of guarana ethanolic extract (GEE) on lifespan using C. elegans. The major known longevity pathways were analyzed through mutant worms and RT-qPCR assay (DAF-2, DAF-16, SKN-1, SIR-2.1, HSF-1). The possible involvement of purinergic signaling was also investigated. This study demonstrated that GEE acts through antioxidant activity, DAF-16, HSF-1, and SKN-1 pathways, and human adenosine receptor ortholog (ADOR-1) to extend lifespan. GEE also downregulated skn-1, daf-16, sir-2.1 and hsp-16.2 in 9-day-old C. elegans, which might reflect less need to activate these protective genes due to direct antioxidant effects. Our results contribute to the comprehension of guarana effects in vivo, which might be helpful to prevent or treat aging-associated disorders, and also suggest purinergic signaling as a plausible therapeutic target for longevity studies.
Assuntos
Antioxidantes/farmacologia , Caenorhabditis elegans/efeitos dos fármacos , Paullinia/química , Extratos Vegetais/farmacologia , Envelhecimento/efeitos dos fármacos , Animais , Antioxidantes/isolamento & purificação , Caenorhabditis elegans/fisiologia , Longevidade/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TempoRESUMO
Guarana (Paullinia cupana) is habitually ingested by people in the Amazon region and is a key ingredient in various energy drinks consumed worldwide. Extension in longevity and low prevalence of chronic age-related diseases have been associated to habitual intake of guarana. Anti-aging potential of guarana was also demonstrated in Caenorhabditis elegans; however, the mechanisms involved in its effects are not clear. Herein, we investigated the putative pathways that regulate the effects of guarana ethanolic extract (GEE) on lifespan using C. elegans. The major known longevity pathways were analyzed through mutant worms and RT-qPCR assay (DAF-2, DAF-16, SKN-1, SIR-2.1, HSF-1). The possible involvement of purinergic signaling was also investigated. This study demonstrated that GEE acts through antioxidant activity, DAF-16, HSF-1, and SKN-1 pathways, and human adenosine receptor ortholog (ADOR-1) to extend lifespan. GEE also downregulated skn-1, daf-16, sir-2.1 and hsp-16.2 in 9-day-old C. elegans, which might reflect less need to activate these protective genes due to direct antioxidant effects. Our results contribute to the comprehension of guarana effects in vivo, which might be helpful to prevent or treat aging-associated disorders, and also suggest purinergic signaling as a plausible therapeutic target for longevity studies.
Assuntos
Animais , Extratos Vegetais/farmacologia , Caenorhabditis elegans/efeitos dos fármacos , Paullinia/química , Antioxidantes/farmacologia , Fatores de Tempo , Envelhecimento/efeitos dos fármacos , Caenorhabditis elegans/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Longevidade/efeitos dos fármacos , Antioxidantes/isolamento & purificaçãoRESUMO
The aim of the study was to investigate the association between Glutathione S-transferase P1 (GSTP1) gene polymorphism with obesity and markers of cardiometabolic risk. A cross-sectional study was carried out in individuals aged≥18 and ≤30 years. The study included 54 normal weight, 27 overweight and 68 obese volunteers. Anthropometric measurements and biochemical parameters were evaluated, the DNA was extracted from blood samples and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to measure GSTP1 Ile105Val gene polymorphism of the study participants. Also, biochemical analysis and hormone assays were carried out. A positive association between GSTP1 polymorphism and obesity was observed on subjects carrying at least one G allele (AG and GG). GG genotype was found only in the obese group. The G allele carriers presented 2.4 times higher chance of obesity when compared to those with the AA genotype. These results were independent of sex and age. We suggest that despite a study in population regional (south of Brazil), the GSTP1 gene polymorphism may play a significant role in the increase of susceptibility of obesity and contribute to identify the cardiovascular risk in young adults.
Assuntos
Alelos , Predisposição Genética para Doença , Glutationa S-Transferase pi/genética , Mutação de Sentido Incorreto , Obesidade/genética , Polimorfismo de Fragmento de Restrição , Adulto , Substituição de Aminoácidos , Feminino , Humanos , Masculino , Obesidade/enzimologia , Adulto JovemRESUMO
Previous investigations suggested that elevated cell-free DNA (cfDNA) can indicate non-healthy states. However, the potential association between cfDNA seminal plasma levels and fertility sperm parameters has not yet been determined. Therefore, the present study evaluated the association between seminal cfDNA levels and sperm fertility criteria to determine the use of seminal cfDNA quantification. An in vivo protocol quantified cfDNA levels of semen samples obtained from 163 male patients using fluorescent PicoGreen dye staining. To confirm if semen cfDNA quantification is realistic, an in vitro complementary test was performed using three or four semen samples. The fresh sperm samples were exposed to paraquat that generates high levels of superoxide anion causing oxidative stress and cell mortality. The results showed significant association between dsDNA levels and several sperm fertility parameters, such as low viability and alterations of motility and morphology. The in vitro analysis confirmed the association between dsDNA levels and sperm viability. Together, these results suggest that dsDNA levels could be an important biomarker to test sperm fertility.
Assuntos
DNA/análise , Corantes Fluorescentes/química , Infertilidade Masculina/fisiopatologia , Análise do Sêmen/métodos , Sêmen/química , Motilidade dos Espermatozoides , Adulto , Sistema Livre de Células , Humanos , Masculino , Compostos Orgânicos/química , Estresse OxidativoRESUMO
Rosuvastatin is a cholesterol-lowering drug that also attenuates the inflammatory process and oxidative stress via the reduction of superoxide anion production. Superoxide anions are metabolized by manganese-dependent superoxide dismutase (MnSOD or SOD2) in the mitochondria. In humans, there is a gene polymorphism where a change of alanine (Ala) to valine (Val) occurs at the 16th amino acid (Ala16Val-SOD2). The VV genotype has been associated with the risk of developing several metabolic diseases, such as hypercholesterolemia. Thus, to further explore this phenomenon, this study investigated the influence of the Val16Ala-SOD2 polymorphism on the lipid profile and inflammatory and fibrinolytic biomarkers of 122 hypercholesterolemic patients undergoing the first pharmacological cholesterol-lowering therapy who were treated with 20 mg rosuvastatin for 120 days. The findings indicate that the VV patients who present a low-efficiency SOD2 enzyme exhibit an attenuated response to rosuvastatin compared with the A-allele patients. The effect of rosuvastatin on inflammatory and fibrinolytic biomarkers was also less intense in the VV patients. These results suggest some pharmacogenetic effects of Val16Ala-SOD2 in hypercholesterolemia treatment.
Assuntos
Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Fibrinólise/efeitos dos fármacos , Fibrinolíticos/uso terapêutico , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Hipercolesterolemia/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Variantes Farmacogenômicos , Polimorfismo de Nucleotídeo Único , Rosuvastatina Cálcica/uso terapêutico , Superóxido Dismutase/genética , Adulto , Idoso , Biomarcadores/sangue , Colesterol/sangue , Resistência a Medicamentos/genética , Feminino , Frequência do Gene , Heterozigoto , Homozigoto , Humanos , Hipercolesterolemia/sangue , Hipercolesterolemia/enzimologia , Hipercolesterolemia/genética , Mediadores da Inflamação/sangue , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Farmacogenética , Fenótipo , Estudos Prospectivos , Fatores de Risco , Superóxidos/metabolismo , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
AIM: To evaluate the in vitro toxicity of irrigating solutions and pharmacological associations used in the pulpectomy of primary teeth. METHODOLOGY: The cell viability (MTT), lipid peroxidation (TBARS), alkaline comet assay and GEMO tests were performed to evaluate the cytotoxicity and genotoxicity of solutions: sodium hypochlorite (1% and 2.5%), 2% chlorhexidine, 6% citric acid and 17% EDTA, which were tested, individually and in association, exposing human peripheral blood mononuclear cells (MTT, TBARS and alkaline comet assay), at 24 and 72 h, and dsDNA (GEMO). After performing the Kolmogorov-Smirnov test, data were analysed by anova followed by Dunnett's post hoc test, and Kruskal-Wallis followed by Dunn post hoc test. A significance level was established at P < 0.05. RESULTS: All irrigating solutions and pharmacological associations reduced cell viability at 24 h (P < 0.05). These reductions were maintained after 72 h, except for EDTA and associations of sodium hypochlorite (1% and 2.5%) with EDTA and of chlorhexidine with EDTA. Lipid peroxidation at 24 h was caused by EDTA and by 2.5% sodium hypochlorite with EDTA; it was also caused at 72 h by sodium hypochlorite (1% and 2.5%) and the three associations with citric acid (P < 0.05). All groups caused DNA damage when assessed by the alkaline comet assay, at 24 h and 72 h (P < 0.05). In the GEMO assay, all groups caused dsDNA damage (P < 0.05), except for chlorhexidine with EDTA. CONCLUSION: All groups showed some level of toxicity. Amongst the main solutions, chlorhexidine presented less cytotoxic potential. EDTA was the least cytotoxic of the auxiliary irrigant solutions, and the association of these two solutions showed the lowest toxicity potential amongst all groups.
Assuntos
DNA/efeitos dos fármacos , Leucócitos Mononucleares/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Pulpectomia/efeitos adversos , Dano ao DNA , Leucócitos Mononucleares/metabolismo , Irrigantes do Canal Radicular , Dente Decíduo , Testes de ToxicidadeRESUMO
AIM: To evaluate the cytotoxicity, oxidative stress and genotoxicity in vitro of four iodoform pastes and three calcium hydroxide pastes. METHODOLOGY: Peripheral blood mononuclear cells (PBMCs) and pure calf thymus DNA (dsDNA) were exposed to extracts of the pastes. Cytotoxicity was assessed with the MTT assay. Generation of reactive oxygen species (ROS) was evaluated using a DCFH-DA assay, and lipid peroxidation was evaluated using a TBARS assay. Genotoxicity was evaluated using the alkaline comet assay and Genomodifier capacity assay (GEMO). All tests were performed after 24 h and 72 h of cell exposure, except GEMO. After performing the Kolmogorov-Smirnov test, data were analysed by Kruskal-Wallis and Dunn's post-tests, and anova with Dunnett's post-test, with a significance level established at P < 0.05. RESULTS: The MTT assay revealed that chlorhexidine, Maxitrol and neomycin sulphate + bacitracin pastes decreased cell viability after 24 h (P < 0.05). No group was associated with a significant decreased cell viability or lipid peroxidation after 72 h. Calcium hydroxide pastes increased the cell viability levels at both experimental times (P < 0.05). Lipid peroxidation was observed with the exposure of cells to calcium hydroxide pastes after 24 h (P < 0.05). Exposure to chlorhexidine, Guedes-Pinto and calcium hydroxide pastes resulted in a significant increase in ROS after 24 h (P < 0.05), whereas iodoform pastes and Calen thickened with zinc oxide significantly increased the ROS after 72 h (P < 0.05). The comet assay revealed that exposure of the PBMCs to iodoform pastes did not damage DNA at either period of time (P > 0.05). However, chlorhexidine paste caused DNA damage in dsDNA (P < 0.05). Calcium hydroxide pastes caused DNA damage in both tests (P < 0.05). CONCLUSION: The pastes varied in their ability to induce cytotoxicity, genotoxicity and oxidative stress. In general, Guedes-Pinto, Maxitrol and neomycin sulphate + bacitracin pastes exhibited better biocompatibility in vitro.
Assuntos
DNA/efeitos dos fármacos , Hidrocarbonetos Iodados/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Materiais Restauradores do Canal Radicular/farmacologia , Análise de Variância , Animais , Bovinos , Dano ao DNA , Humanos , Leucócitos Mononucleares/metabolismo , Teste de Materiais , Espécies Reativas de Oxigênio/metabolismo , Estatísticas não Paramétricas , Dente Decíduo , Testes de ToxicidadeRESUMO
Hypercholesterolemia is a metabolic disorder characterized by high levels of low-density lipoprotein and blood cholesterol, causing inflammatory lesion. Purinergic signaling modulates the inflammatory and immune responses through adenine nucleotides and nucleoside. Guaraná has hypocholesterolemic and antiinflammatory properties. Considering that there are few studies demonstrating the effects of guaraná powder on the metabolism of adenine nucleotides, we investigated its effects on the activity of ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) and ecto-adenosine deaminase activity in lymphocytes of rats with diet-induced hypercholesterolemia. The rats were divided into hypercholesterolemic and normal diet groups. Each group was subdivided by treatment: saline, guaraná powder 12.5, 25, or 50 mg/kg/day and caffeine concentration equivalent to highest dose of guaraná, fed orally for 30 days. An increase in adenosine triphosphate hydrolysis was observed in the lymphocytes of rats with hypercholesterolemia and treated with 25 or 50 mg/kg/day when compared with the other groups. The hypercholesterolemic group treated with the highest concentration of guaraná powder showed decreased ecto-adenosine deaminase activity compared with the normal diet groups. Guaraná was able to reduce the total cholesterol and low-density lipoprotein cholesterol to basal levels in hypercholesterolemic rats. High concentrations of guaraná associated with a hypercholesterolemic diet are likely to have contributed to the reduction of the inflammatory process.
Assuntos
Cafeína/farmacologia , Hipercolesterolemia/tratamento farmacológico , Paullinia/química , Teobromina/farmacologia , Teofilina/farmacologia , Adenosina Desaminase/metabolismo , Animais , Colesterol/sangue , LDL-Colesterol/sangue , Dieta Hiperlipídica , Linfócitos/enzimologia , Masculino , Preparações de Plantas/farmacologia , Ratos , Ratos WistarRESUMO
Os problemas relacionados ao armazenamento vesical são muitos e relevantes. Eles, além de influírem de forma efetiva na qualidade de vida, podem eventualmente evoluir para falência renal. Existem vários trabalhos, os quais descrevem as propriedades imunomoduladoras e imunossupressoras das células-tronco mesenquimais derivadas do tecido adiposo (ADSCs). Objetiva-se com o presente avaliar clínica, ecográfica e anatomofisiologicamente o alotransplante parcial de bexiga a fresco em coelhos, utilizando como agente imunomodulador ADSCs alogênicas. Para isso foram utilizados 25 coelhos, sendo um deles macho e doador das ADSCs, e os outros 24 eram fêmeas, submetidas a alotransplante parcial de bexiga, sendo tratadas com ciclosporina (GCi) ou células-tronco mesenquimais (GCe). Conclui-se que as ADSCs foram suficientes para evitar sinais clínicos e ecográficos de rejeição ao alotransplante de vesícula urinária, mantendo a estrutura anatomofisiológica vesical por até 30 dias em coelhos.
The problems related to bladder storage are many and significant. In addition to effectively impacting the quality of life, they can eventually progress to kidney failure. There are several studies which describe the immunomodulatory and immunosuppressive properties of ADSCs. The aim of this study is to clinically, through sonography, anatomically and physiologically evaluate fresh partial allograft bladder from rabbits using allogeneic ADSCs as immunomodulator agents. For such, 25 rabbits were used, one being a male ADSCs donor, and the other 24 females who underwent simultaneous partial allograft bladder, being treated with cyclosporine (GCi) or mesenchymal stem cells (GCe). It was concluded that ADSCs were sufficient to prevent clinical and ultrasound signs of allograft rejection of the urinary bladder. These bladders retained the anatomophysiological structure for 30 days in rabbits.