RESUMO
Coronary artery endothelial cells (CAEC) exert an important role in the development of cardiovascular disease. Dysfunction of CAEC is associated with cardiovascular disease in subjects with type 2 diabetes mellitus (T2DM). However, comprehensive studies of the effects that a diabetic environment exerts on this cellular type are scarce. The present study characterized the molecular perturbations occurring on cultured bovine CAEC subjected to a prolonged diabetic environment (high glucose and high insulin). Changes at the metabolite and peptide level were assessed by Liquid Chromatography-Mass Spectrometry (LC-MS2) and chemoinformatics. The results were integrated with published LC-MS2-based quantitative proteomics on the same in vitro model. Our findings were consistent with reports on other endothelial cell types and identified novel signatures of DNA/RNA, amino acid, peptide, and lipid metabolism in cells under a diabetic environment. Manual data inspection revealed disturbances on tryptophan catabolism and biosynthesis of phenylalanine-based, glutathione-based, and proline-based peptide metabolites. Fluorescence microscopy detected an increase in binucleation in cells under treatment that also occurred when human CAEC were used. This multi-omics study identified particular molecular perturbations in an induced diabetic environment that could help unravel the mechanisms underlying the development of cardiovascular disease in subjects with T2DM.
Assuntos
Doenças Cardiovasculares , Diabetes Mellitus Tipo 2 , Aminoácidos/metabolismo , Animais , Doenças Cardiovasculares/complicações , Bovinos , DNA/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Células Endoteliais/metabolismo , Humanos , Metabolismo dos Lipídeos , Peptídeos/metabolismo , RNA/metabolismoRESUMO
We evaluated the effects of a non-hepatotropic parasite infection (Taenia crassiceps) on the outcome of acetaminophen-induced acute liver failure in mice. Uninfected and T. crassiceps infected mice orally received either 300 mg/kg acetaminophen or water as vehicle (n = 5 per group). Survival analysis, hepatocyte necrosis, alanine aminotransferase (ALT) levels, CYP2E1 protein, interleukin (IL-) 5, and IL-6 were assessed for all groups. All infected mice died within 16 h after exposure to acetaminophen (Tc+APAP group), whereas only one-third of uninfected animals exposed to acetaminophen (APAP group) died. Uninfected (Control group) and infected (Tc group) mice that received the vehicle showed no liver damage. Tc+APAP mice exhibited massive liver necrosis characterised by marked balloning degeneration of hepatocytes and higher serum ALT compared to Control, Tc, and APAP animals. Liver tissue from Tc+APAP mice also displayed increased expression of CYP2E1 protein and higher mRNA and protein levels of IL-5 and IL-6 compared to the other groups. These findings suggest that non-hepatotropic parasite infections may increase mortality following acute liver failure by promoting hepatocyte necrosis via IL-5 and IL-6-dependent CYP2E1 overproduction. This study identifies new potential risk factors associated with severe acute liver failure in patients.
Assuntos
Animais , Feminino , Acetaminofen , Analgésicos não Narcóticos , Falência Hepática Aguda , Teníase/parasitologia , Acetaminofen/administração & dosagem , Alanina Transaminase/sangue , Analgésicos não Narcóticos/administração & dosagem , Biomarcadores/sangue , Citocromo P-450 CYP2E1/biossíntese , Citocromo P-450 CYP2E1/sangue , Modelos Animais de Doenças , Hepatócitos/parasitologia , Hepatócitos/patologia , Interleucina-5/sangue , Interleucina-6/sangue , Falência Hepática Aguda/induzido quimicamente , Falência Hepática Aguda/mortalidade , Falência Hepática Aguda/parasitologia , Falência Hepática Aguda/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Teníase/patologiaRESUMO
We evaluated the effects of a non-hepatotropic parasite infection (Taenia crassiceps) on the outcome of acetaminophen-induced acute liver failure in mice. Uninfected and T. crassiceps infected mice orally received either 300 mg/kg acetaminophen or water as vehicle (n = 5 per group). Survival analysis, hepatocyte necrosis, alanine aminotransferase (ALT) levels, CYP2E1 protein, interleukin (IL-) 5, and IL-6 were assessed for all groups. All infected mice died within 16 h after exposure to acetaminophen (Tc+APAP group), whereas only one-third of uninfected animals exposed to acetaminophen (APAP group) died. Uninfected (Control group) and infected (Tc group) mice that received the vehicle showed no liver damage. Tc+APAP mice exhibited massive liver necrosis characterised by marked balloning degeneration of hepatocytes and higher serum ALT compared to Control, Tc, and APAP animals. Liver tissue from Tc+APAP mice also displayed increased expression of CYP2E1 protein and higher mRNA and protein levels of IL-5 and IL-6 compared to the other groups. These findings suggest that non-hepatotropic parasite infections may increase mortality following acute liver failure by promoting hepatocyte necrosis via IL-5 and IL-6-dependent CYP2E1 overproduction. This study identifies new potential risk factors associated with severe acute liver failure in patients.
Assuntos
Acetaminofen , Analgésicos não Narcóticos , Falência Hepática Aguda , Teníase/parasitologia , Acetaminofen/administração & dosagem , Alanina Transaminase/sangue , Analgésicos não Narcóticos/administração & dosagem , Animais , Biomarcadores/sangue , Citocromo P-450 CYP2E1/biossíntese , Citocromo P-450 CYP2E1/sangue , Modelos Animais de Doenças , Feminino , Hepatócitos/parasitologia , Hepatócitos/patologia , Interleucina-5/sangue , Interleucina-6/sangue , Falência Hepática Aguda/induzido quimicamente , Falência Hepática Aguda/mortalidade , Falência Hepática Aguda/parasitologia , Falência Hepática Aguda/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Teníase/patologiaRESUMO
Lung cancer is one of the most common types of cancer in men and women and a leading cause of death worldwide resulting in more than one million deaths per year. The venom of marine snails Conus contains up to 200 pharmacologically active compounds that target several receptors in the cell membrane. Due to their diversity and specific binding properties, Conus toxins hold great potential as source of new drugs against cancer. We analyzed the cytotoxic effect of a 17-amino acid synthetic peptide (s-cal14.1a) that is based on a native toxin (cal14.1a) isolated from the sea snail Conus californicus. Cytotoxicity studies in four lung cancer cell lines were complemented with measurement of gene expression of apoptosis-related proteins Bcl-2, BAX and the pro-survival proteins NFκB-1 and COX-2, as well as quantification of caspase activity. Our results showed that H1299 and H1437 cell lines treated with s-call4.1a had decreased cell viability, activated caspases, and reduced expression of the pro-survival protein NFκB-1. To our knowledge, this is the first report describing activation of apoptosis in human lung cancer cell lines by s-cal14.1a and we offer insight into the possible mechanism of action.
Assuntos
Antineoplásicos/farmacologia , Venenos de Moluscos/farmacologia , Peptídeos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 7/metabolismo , Linhagem Celular Tumoral , Caramujo Conus , Ciclo-Oxigenase 2/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , NF-kappa B/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Mensageiro/metabolismo , Proteína X Associada a bcl-2/genéticaRESUMO
Pharmacological concentrations of biotin have pleiotropic effects. Several reports have documented that biotin supplementation decreases hyperglycemia. We have shown that a biotin-supplemented diet increased insulin secretion and the mRNA abundance of proteins regulating insulin transcription and secretion. We also found enlarged pancreatic islets and modified islet morphology. Other studies have shown that pharmacological concentrations of biotin modify tissue structure. Although biotin administration is considered safe, little attention has been given to its effect on tissue structure. In this study, we investigated the effect of biotin supplementation on hepatic morphology and liver toxicity markers. Male BALB/cAnN Hsd mice were fed a control or a biotin-supplemented diet for 8 weeks. Versus the control mice, biotin-supplemented mice had an altered portal triad with dilated sinusoids, increased vascularity, and bile conducts. Furthermore, we observed an increased proportion of nucleomegaly and binucleated hepatocytes. In spite of the liver morphological changes, no differences were observed in the serum liver damage indicators, oxidative stress markers, or antioxidant enzymes. Our data demonstrate for the first time that biotin supplementation affects liver morphology in normal mice, and that these modifications are not paralleled with damage markers.
Assuntos
Biotina/farmacologia , Suplementos Nutricionais , Hepatócitos , Hepatopatias , Fígado , Animais , Biomarcadores/metabolismo , Hepatócitos/metabolismo , Hepatócitos/patologia , Fígado/lesões , Fígado/metabolismo , Fígado/patologia , Hepatopatias/metabolismo , Hepatopatias/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
In this work we studied the influence of sex hormones on heart and mitochondrial functions, from adult castrated female and male, and intact rats. Castration was performed at their third week of life and on the fourth month animals were subjected to heart ischemia and reperfusion. Electrocardiogram and blood pressure recordings were made, cytokines levels were measured, histopathological studies were performed and thiobarbituric acid reactive species were determined. At the mitochondrial level respiratory control, transmembranal potential and calcium management were determined; Western blot of some mitochondrial components was also performed. Alterations in cardiac function were worst in intact males and castrated females as compared with those found in intact females and castrated males, cytokine levels were modulated also by hormonal status. Regarding mitochondria, in those obtained from hearts from castrated females without ischemia-reperfusion, all evaluated parameters were similar to those observed in mitochondria after ischemia-reperfusion. The results show hormonal influences on the heart at functional and mitochondrial levels.
Assuntos
Coração/fisiopatologia , Mitocôndrias Cardíacas/fisiologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Animais , Castração , Citocromos c/metabolismo , Citocinas/metabolismo , Estradiol/sangue , Feminino , Masculino , Traumatismo por Reperfusão Miocárdica/sangue , Miocárdio/metabolismo , Miocárdio/patologia , Ratos , Ratos Sprague-Dawley , Receptores de Estradiol/metabolismo , Caracteres Sexuais , Testosterona/sangue , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Recent evidence suggests that hormonal effects on mitochondria could be mediated by mitochondrial estrogen receptors (mtERs). These receptors are new candidates for the beneficial estrogenic effects on mitochondria in different physiological conditions. The aim of this investigation was to study mtER expression during brain aging. We analyzed mtERalpha and mtERbeta expression in cortical, hippocampal and hypothalamic mitochondria of young adult (3months) and aged (18 months) female Wistar rats by Western blot. In addition, we explored the interaction of mtERbeta with respiratory complex V by using coimmunoprecipitation assays. The results show that mtERalpha and mtERbeta are present in young and aged brain mitochondria. We also demonstrate that mtERs are expressed as variants and have a brain region specific distribution. The predominant mtER variants detected were of 61 and 55KDa for mtERalpha and of 63 and 52KDa for mtERbeta. However, we did not observe differences in the mtERalpha or beta content between the two age groups studied. Additionally, we show that mtERbeta interacts with complex V. The overall results demonstrate that there is a differential expression of mtERalpha and mtERbeta variants in different brain areas, indicating that they may participate in different functions in the brain during aging.