RESUMO
Obesity is associated with an increased incidence and aggressiveness of breast cancer and is estimated to increment the development of this tumor by 50 to 86%. These associations are driven, in part, by changes in the serum molecules. Epidemiological studies have reported that Metformin reduces the incidence of obesity-associated cancer, probably by regulating the metabolic state. In this study, we evaluated in a breast cancer in-vitro model the activation of the IR-ß/Akt/p70S6K pathway by exposure to human sera with different metabolic and hormonal characteristics. Furthermore, we evaluated the effect of brief Metformin treatment on sera of obese postmenopausal women and its impact on Akt and NF-κB activation. We demonstrated that MCF-7 cells represent a robust cellular model to differentiate Akt pathway activation influenced by the stimulation with sera from obese women, resulting in increased cell viability rates compared to cells stimulated with sera from normal-weight women. In particular, stimulation with sera from postmenopausal obese women showed an increase in the phosphorylation of IR-ß and Akt proteins. These effects were reversed after exposure of MCF-7 cells to sera from postmenopausal obese women with insulin resistance with Metformin treatment. Whereas sera from women without insulin resistance affected NF-κB regulation. We further demonstrated that sera from post-Metformin obese women induced an increase in p38 phosphorylation, independent of insulin resistance. Our results suggest a possible mechanism in which obesity-mediated serum molecules could enhance the development of luminal A-breast cancer by increasing Akt activation. Further, we provided evidence that the phenomenon was reversed by Metformin treatment in a subgroup of women.
Assuntos
Neoplasias da Mama , Resistência à Insulina , Menopausa , Proteínas Proto-Oncogênicas c-akt , Neoplasias da Mama/patologia , Proliferação de Células , Sobrevivência Celular , Feminino , Humanos , Técnicas In Vitro , Metformina/farmacologia , NF-kappa B , Obesidade/complicações , Proteínas Proto-Oncogênicas c-akt/metabolismo , Soro/efeitos dos fármacos , Soro/metabolismoRESUMO
We investigated the ability of tannic acid (TA) to prevent oxidative and nitrosative damage in the brain, liver, kidney, and serum of a rat model of acute hypermethioninemia. Young Wistar rats were divided into four groups: I (control), II (TA 30 mg/kg), III (methionine (Met) 0.4 g/kg + methionine sulfoxide (MetO) 0.1 g/kg), and IV (TA/Met + MetO). Rats in groups II and IV received TA orally for seven days, and rats of groups I and III received an equal volume of water. After pretreatment with TA, rats from groups II and IV received a single subcutaneous injection of Met + MetO, and were euthanized 3 h afterwards. In specific brain structures and the kidneys, we observed that Met + MetO led to increased reactive oxygen species (ROS), nitrite, and lipid peroxidation levels, followed by a reduction in thiol content and antioxidant enzyme activity. On the other hand, pretreatment with TA prevented both oxidative and nitrosative damage. In the serum, Met + MetO caused a decrease in the activity of antioxidant enzymes, which was again prevented by TA pretreatment. In contrast, in the liver, there was a reduction in ROS levels and an increase in total thiol content, which was accompanied by a reduction in catalase and superoxide dismutase activities in the Met + MetO group, and pretreatment with TA was able to prevent only the reduction in catalase activity. Conclusively, pretreatment with TA has proven effective in preventing oxidative and nitrosative changes caused by the administration of Met + MetO, and may thus represent an adjunctive therapeutic approach for treatment of hypermethioninemia.
Assuntos
Erros Inatos do Metabolismo dos Aminoácidos/tratamento farmacológico , Glicina N-Metiltransferase/deficiência , Estresse Nitrosativo/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Taninos/farmacologia , Erros Inatos do Metabolismo dos Aminoácidos/metabolismo , Erros Inatos do Metabolismo dos Aminoácidos/patologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Glutationa Peroxidase/genética , Glicina N-Metiltransferase/metabolismo , Humanos , Rim/efeitos dos fármacos , Rim/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Estresse Nitrosativo/genética , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/genética , Ratos , Espécies Reativas de Oxigênio/metabolismo , Soro/efeitos dos fármacos , Soro/metabolismo , Superóxido Dismutase/genéticaRESUMO
Introducción: Las interferencias en el proteinograma electroforético por electroforesis capilar incluyen la aparición de picos con concentraciones y movilidades electroforéticas, que podrían simular la presencia de un componente monoclonal. Objetivos: Ante la aparición de un pico adicional con movilidad intera2-ß por electroforesis capilar (Minicap®-Sebia), el objetivo fue identificar el interferente y evaluar su relación con la funcionalidad renal. Material y métodos: Se estudiaron muestras de suero que presentaron dicha interferencia en un período de un año mediante proteinograma en soporte sólido, electroinmunofijación e inmunoelectroforesis. Se adicionó in vitro el probable interferente para confirmar su movilidad electroforética. Se evaluó el impacto de la corrección de la interferencia con la herramienta "eliminación de artefactos" (Phoresis®-Sebia) y la correlación de la concentración del pico a línea de base del interferente con la estimación de la tasa de filtrado glomerular (CKD- EPI). Resultados: La integración a la línea de base de los picos fue de 0,07-0,36 g/dL. No se observaron particularidades al realizar los estudios complementarios. Se evidenció, en todos los casos, la administración de iopamidol como medio de contraste, confirmándose su movilidad electroforética por su adición in vitro. Mediante la herramienta "eliminación de artefactos" se recuperaron los niveles basales de las fracciones. Se demostró la existencia de una correlación entre la concentración del pico a línea de base del interferente y la estimación de la tasa de filtración glomerular por CKD-EPI (r=-0.534, p<0.0001). Conclusiones: Se identificó al interferente como Iopamidol y se demostró su relación con la disminución de la tasa de filtración glomerular
Introduction: Interferences in the electrophoretic proteinogram by capillary electrophoresis include the appearance of peaks with concentrations and electrophoretic mobilities, which could simulate the presence of a monoclonal component. Objectives: In the light of an additional peak with interα2-ß mobility by capillary electrophoresis (MINICAP®-Sebia), the aim was to identify the interferent and evaluate its connection to renal functionality. Methods: Serum samples that presented this interference over a period of one year were studied by proteinogram on solid support, electroimmunofixation and immunoelectrophoresis. The probable interferent was added in vitro to confirm its electrophoretic mobility. The impact of the interference correction with the "artifact removal" tool (Phoresis®-Sebia) and the correlation of the baseline peak concentration of the interferent with the estimation of the glomerular filtration rate (CKD-EPI) were evaluated. Results: The integration to the baseline of the peaks was 0.07-0.36 g/dL. No particularities were observed when performing the complementary studies. In all cases, the administration of Iopamidol as a contrast medium was demonstrated, confirming its electrophoretic mobility due to its in vitro addition. Using the "artifact removal" tool, the basal levels of the fractions were recovered. The existence of a correlation between the concentration of the baseline peak of the interferent and the estimation of the glomerular filtration rate by CKD-EPI was shown (r=-0.534, p <0.0001). Conclusions: The interferent was identified as Iopamidol and its connection to the decrease in the glomerular filtration rate was demonstrated
Assuntos
Humanos , Iopamidol , Proteínas Sanguíneas , Eletroforese Capilar , Meios de Contraste , Imunoeletroforese , Soro/efeitos dos fármacos , Barreira de Filtração GlomerularRESUMO
Diabetes Mellitus (DM) is associated with an increased susceptibility to various infections, which might be attributed to changes in immune response owing to chronic hyperglycemia. Nucleoside triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase, and adenosine deaminase (ADA) are important enzymes involved in the generation of anti-aggregant and anti-inflammatory microenvironments. The aim of this study was to evaluate the effect of gallic acid (GA) on the hematological parameters and ectonucleotidase activities in platelets, lymphocytes, and serum of diabetic rats. Experimental rats were categorized into 4 groups: (i) control -saline, (ii) control - GA, (iii) diabetic -saline, and (iv) diabetic - GA. One week after induction of DM using streptozotocin (65â¯mg/kg), GA (30â¯mg/kg) or saline was orally administered to the rats for 21 days. Our results demonstrated that the concentration of mean corpuscular hemoglobin was decreased, whereas that of red cell distribution was increased in the diabetic group, however, GA could revert these alterations. Moreover, in diabetic rats, GA reverted the increase in ATP and ADP hydrolysis and ADA activity in lymphocytes, and it prevented the increase in NTPDase and ADA activities in platelets. A decrease in ATP hydrolysis and an increase in ADP and AMP hydrolysis were observed in the serum of diabetic rats; however, GA treatment could solely revert changes in ATP hydrolysis. Our study suggests that GA exhibits beneficial effects on immuno- and thrombo-regulatory responses in DM and that these effects may be related to the modulation of purinergic signaling.
Assuntos
Plaquetas/metabolismo , Diabetes Mellitus Experimental/sangue , Ácido Gálico/farmacologia , Linfócitos/metabolismo , Nucleotídeos de Purina/metabolismo , Soro/metabolismo , Animais , Plaquetas/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Ácido Gálico/uso terapêutico , Linfócitos/efeitos dos fármacos , Masculino , Agregação Plaquetária/efeitos dos fármacos , Agregação Plaquetária/fisiologia , Nucleotídeos de Purina/agonistas , Nucleotídeos de Purina/antagonistas & inibidores , Ratos , Ratos Wistar , Soro/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologiaRESUMO
OBJETIVO: evaluar el comportamiento de las queratitis bacterianas con el tratamiento coadyuvante de suero autólogo tópico al 50 %. MÉTODOS: estudio comparativo, longitudinal y prospectivo. La muestra estuvo constituida por 60 pacientes, divididos de forma aleatoria en dos grupos: A) utilizó tratamiento antibiótico convencional y suero autólogo y B)utilizó solo tratamiento antibiótico convencional (cefazolina y amikacina). Los datos almacenados se procesaron utilizando el paquete estadístico SPSS 15. Las variables se expresaron según sus respectivas medidas de resumen y para la comparación de las terapias se aplicaron pruebas de hipótesis, con un nivel de confianza del 95 % y de error inferior al 0,05 %. RESULTADOS: predominaron en los aislamientos microbiológicos Staphylococcus y Pseudomonas; seguidas por los Streptococcus, gonococos y enterobacterias. En cuanto al tiempo de aparición de los signos que favorecen la cicatrización corneal y la respuesta terapéutica, encontramos que con la aplicación tópica del suero autólogo, en casi dos tercios de los pacientes, estos se manifestaron a partir de la segunda semana de tratamiento y se obtuvo una respuesta terapéutica favorable. CONCLUSIONES: el suero autólogo al 50 % resulta ser un complemento terapéutico efectivo en el manejo de las queratitis infecciosas de etiología bacteriana.
OBJECTIVE: to evaluate the behavior of bacterial keratitis with the adjuvant 50% topical autologous serum treatment. METHODS: comparative, longitudinal and prospective study of 60 patients, randomly distributed into two groups. Group A used the conventional antibiotics treatment plus autologous serum and Group B used only conventional antibiotics treatment (cephazolin and amykacin). The collected data were processed by the statistical package SPSS15. The variables were expressed according to their respective summary measures and hypothesis tests were applied to compare the two therapies; a confidence level of 95 % and error level less than 0,05 % were used. RESULTS: in the microbiological isolates, Staphylococcus and Pseudomonas prevailed, followed by Streptococcus, gonococci and Enterobacteriaceae. Regarding the time of onset of the signs that favor corneal healing and the therapeutic response, it was found that with the topical application of the autologous serum, almost two thirds of the patients showed healing signs since the second week of treatment, revealing a favorable therapeutic response. CONCLUSIONS: the 50 % autologous serum proves to be an effective therapeutic complement in the management of the infectious keratitis of bacterial etiology.
Assuntos
Humanos , Soluções Oftálmicas , Úlcera da Córnea/diagnóstico , Úlcera da Córnea/terapia , Soro/efeitos dos fármacos , Ceratite/terapia , Estudos de Casos e Controles , Estudos Prospectivos , Estudos LongitudinaisRESUMO
PURPOSE: To study the influence of albumin on changes of liver function in the extrahepatic biliary obstruction through an experimental model in rats. METHODS: Sixty rats were divided into four groups: Group C (Control): 6 animals. Group M (Fictitious Operation): 18 rats underwent laparotomy and handling of the bile ducts; Groups O (extrahepatic biliary obstruction) and A (Treated with 2% albumin): 18 animals in each group underwent ligation of the ductus liver; The animals in groups M, O and A were divided into three subgroups of 6 animals each to be killed in the 7, 14 and 21 days postoperative (POD). Blood was drawn for determination of total bilirubin (TB), indirect bilirubin (IB), direct bilirubin (DB), alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). RESULTS: On POD 7, BI levels were 4.5 mg / dl in group O and 2.1 mg / dl in group A (p = 0.025). On the 14th POD, the levels of PA were 1185.2 U / l in the group and O 458.3 U / l in group A (p = 0.004). ALT levels were 101.7 U / l in the group O and 75.7 U / l in group A (= 0.037). On POD 21, the levels of ALP were 1069.5 U / l in the group O and 468.3 U / l in group A (p = 0, 004). CONCLUSION: The administration of albumin reduced the serum levels of bilirubin in the 7th day of supplementation.
Assuntos
Albuminas/farmacologia , Ductos Biliares Extra-Hepáticos , Colestase Extra-Hepática/tratamento farmacológico , Fígado/efeitos dos fármacos , Alanina Transaminase/sangue , Fosfatase Alcalina/sangue , Animais , Aspartato Aminotransferases/sangue , Bilirrubina/sangue , Colestase Extra-Hepática/enzimologia , Colestase Extra-Hepática/etiologia , Modelos Animais de Doenças , Laparotomia , Ligadura/métodos , Fígado/enzimologia , Masculino , Ratos , Ratos Wistar , Soro/efeitos dos fármacos , Resultado do TratamentoRESUMO
PURPOSE: To study the influence of albumin on changes of liver function in the extrahepatic biliary obstruction through an experimental model in rats. METHODS: Sixty rats were divided into four groups: Group C (Control): 6 animals. Group M (Fictitious Operation): 18 rats underwent laparotomy and handling of the bile ducts; Groups O (extrahepatic biliary obstruction) and A (Treated with 2 percent albumin): 18 animals in each group underwent ligation of the ductus liver; The animals in groups M, O and A were divided into three subgroups of 6 animals each to be killed in the 7, 14 and 21 days postoperative (POD). Blood was drawn for determination of total bilirubin (TB), indirect bilirubin (IB), direct bilirubin (DB), alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). RESULTS: On POD 7, BI levels were 4.5 mg / dl in group O and 2.1 mg / dl in group A (p = 0.025). On the 14th POD, the levels of PA were 1185.2 U / l in the group and O 458.3 U / l in group A (p = 0.004). ALT levels were 101.7 U / l in the group O and 75.7 U / l in group A (= 0.037). On POD 21, the levels of ALP were 1069.5 U / l in the group O and 468.3 U / l in group A (p = 0, 004). CONCLUSION: The administration of albumin reduced the serum levels of bilirubin in the 7th day of supplementation.
OBJETIVO: Estudar a influência da albumina em alterações funcionais do fígado na obstrução biliar extra-hepática por meio de um modelo experimental desenvolvido em ratos. MÉTODOS: 60 ratos distribuídos em quatro grupos: Grupo C (Controle): 6 animais. Grupo M (Operação Fictícia): 18 ratos submetidos à laparotomia e manuseio das vias biliares; Grupos O (Obstrução Biliar Extra-hepática) e A (Tratados com albumina a 2 por cento): 18 animais, em cada grupo, submetidos à ligadura do ducto hepático; Os animais dos grupos M, O e A foram distribuídos em três subgrupos de 6 animais cada, para serem mortos nos 7°, 14° e 21° dias pós- operatórios (DPO). Foi colhido sangue para dosagem de bilirrubina total (BT), bilirrubina indireta (BI), bilirrubina direta (BD), fosfatase alcalina (FAL), aspartato aminotransferase (AST) e alanina aminotransferase (ALT). RESULTADOS: no 7º DPO, os níveis de BI foram 4,5 mg/dl no grupo O e 2,1mg/dl no grupo A (p=0,025). No 14º DPO, os níveis de FAL foram 1185,2 U/l no grupo O e 458,3 U/l no grupo A (p=0,004). Os níveis de ALT foram de 101,7 U/l no grupo O e 75,7 U/l no grupo A (=0,037). No 21º DPO, os níveis de FAL foram de 1069,5 U/l no grupo Oe de 468,3 U/l no grupo A (p =0, 004). CONCLUSÃO: a administração de albumina reduziu os níveis séricos de bilirrubina indireta no 7°dia de suplementação.
Assuntos
Animais , Masculino , Ratos , Albuminas/farmacologia , Ductos Biliares Extra-Hepáticos , Colestase Extra-Hepática/tratamento farmacológico , Fígado/efeitos dos fármacos , Alanina Transaminase/sangue , Fosfatase Alcalina/sangue , Aspartato Aminotransferases/sangue , Bilirrubina/sangue , Colestase Extra-Hepática/enzimologia , Colestase Extra-Hepática/etiologia , Modelos Animais de Doenças , Laparotomia , Ligadura/métodos , Fígado/enzimologia , Ratos Wistar , Soro/efeitos dos fármacos , Resultado do TratamentoRESUMO
The objective of this study was to verify the effect of the organochalcogen 3-butyl-1-phenyl-2-(phenyltelluro)oct-en-1-one on some parameters of oxidative stress in human serum. Serum of volunteers were incubated for 30 min in the presence or absence of 1, 10, or 30 microM of 3-butyl-1-phenyl-2-(phenyltelluro)oct-en-1-one and oxidative stress was measured. First, we tested the influence of the compound on 1,1-diphenyl-2-picrylhydrazyl (DPPH(*)) radical-scavenging and verified that the organotellurium did not have any antioxidant properties. The organochalcogen was capable to enhance TBARS but the compound was not able to alter carbonyl assay. Furthermore, the organochalcogen provoked a reduction of protein thiol groups measured by the sulfhydryl assay. Moreover, the organotellurium enhanced the activity of catalase and superoxide dismutase, inhibited the activity of glutathione peroxidase and did not modify the glutathione S-transferase activity. Furthermore, nitric oxide production and hydroxyl radical activity were not affected by the compound. Our findings showed that this organochalcogen induces oxidative stress in human serum, indicating that this compound is potentially toxic to human beings.
Assuntos
Compostos Organometálicos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Soro/efeitos dos fármacos , Telúrio/química , Compostos de Bifenilo , Catalase/metabolismo , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Humanos , Radical Hidroxila/metabolismo , Técnicas In Vitro , Indicadores e Reagentes/farmacologia , Óxido Nítrico/metabolismo , Picratos , Carbonilação Proteica , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
In the present study we investigated the effect of homocysteine administration, the main metabolite accumulating in homocystinuria, on cholinesterase activity in rat and human serum. For the in vivo study, 8-, 15- and 60-day-old rats received one subcutaneous injection of homocysteine (0.3, 0.4 or 0.6 micromol/g of body weight, respectively) or saline (control) and were sacrificed 1h later, when serum was collected in order to determine cholinesterase activity. For the in vitro studies, serum of 8-, 15- and 60-day-old untreated rats or 20-25- and 52-60-day-old human beings (healthy volunteers) were incubated with 10-500 microM homocysteine. Results showed that acute hyperhomocysteinemia (in vivo study) significantly reduced cholinesterase activity in the serum of rats of all ages tested. We also observed that 500 microM homocysteine added to the incubation medium (in vitro study) significantly inhibited cholinesterase activity both in serum of rats and humans. Our findings seem to reinforce the proposed associations of cholinesterase activity with hyperhomocysteinemia.