RESUMO
Chronic low-grade vascular inflammation and endothelial dysfunction significantly contribute to the pathogenesis of cardiovascular diseases. In endothelial cells (ECs), anti-inflammatory or pro-inflammatory signaling can be induced by different patterns of the fluid shear stress (SS) exerted by blood flow on ECs. Laminar blood flow with high magnitude is anti-inflammatory, while disturbed flow and laminar flow with low magnitude is pro-inflammatory. Endothelial mechanosensors are the key upstream signaling proteins in SS-induced pro- and anti-inflammatory responses. Being transmembrane proteins, mechanosensors, not only experience fluid SS but also become regulated by the biomechanical properties of the lipid bilayer and the cytoskeleton. We review the apparent effects of pro-inflammatory factors (hypoxia, oxidative stress, hypercholesterolemia, and cytokines) on the biomechanics of the lipid bilayer and the cytoskeleton. An analysis of the available data suggests that the formation of a vicious circle may occur, in which pro-inflammatory cytokines enhance and attenuate SS-induced pro-inflammatory and anti-inflammatory signaling, respectively.
RESUMO
Changes in plasma membrane curvature and intracellular ionic strength are two key features of cell volume perturbations. In this hypothesis we present a model of the responsible molecular apparatus which is assembled of two molecular motors [non-muscle myosin II (NMMII) and protrusive actin polymerization], a spring [a complex between the plasma membrane (PM) and the submembrane actin-based cytoskeleton (smACSK) which behaves like a viscoelastic solid] and the associated signaling proteins. We hypothesize that this apparatus senses changes in both the plasma membrane curvature and the ionic strength and in turn activates signaling pathways responsible for regulatory volume increase (RVI) and regulatory volume decrease (RVD). During cell volume changes hydrostatic pressure (HP) changes drive alterations in the cell membrane curvature. HP difference has opposite directions in swelling versus shrinkage, thus allowing distinction between them. By analogy with actomyosin contractility that appears to sense stiffness of the extracellular matrix we propose that NMMII and actin polymerization can actively probe the transmembrane gradient in HP. Furthermore, NMMII and protein-protein interactions in the actin cortex are sensitive to ionic strength. Emerging data on direct binding to and regulating activities of transmembrane mechanosensors by NMMII and actin cortex provide routes for signal transduction from transmembrane mechanosensors to cell volume regulatory mechanisms.
Assuntos
Citoesqueleto de Actina/metabolismo , Membrana Celular/metabolismo , Tamanho Celular , Miosina Tipo II/metabolismo , Transdução de Sinais , Actomiosina/metabolismo , Animais , Humanos , Pressão HidrostáticaRESUMO
Endothelial cells (ECs) are exposed to molecular dioxygen and its derivative reactive oxygen species (ROS). ROS are now well established as important signaling messengers. Excessive production of ROS, however, results in oxidative stress, a significant contributor to the development of numerous diseases. Here, we analyze the experimental data and theoretical concepts concerning positive pro-survival effects of ROS on signaling pathways in endothelial cells (ECs). Our analysis of the available experimental data suggests possible positive roles of ROS in induction of pro-survival pathways, downstream of the Gi-protein-coupled receptors, which mimics insulin signaling and prevention or improvement of the endothelial dysfunction. It is, however, doubtful, whether ROS can contribute to the stabilization of the endothelial barrier.
RESUMO
BACKGROUND: Calcium, vitamin D and insulin resistance are linked to osteoporosis and cardiovascular disease in menopause. OBJECTIVE: Determine if hemorheological parameters related to blood viscosity in microcirculation are linked to calcium metabolism and insulin resistance in menopause. METHODS: 25-Hydroxyvitamin D (25(OH)D)), 1, 25-dihydroxyvitamin D3 (1, 25(OH)2D), parathyroid hormone, ionized calcium, glucose, insulin and hemoglobin A1c were measured in blood from 43 volunteers. Red blood cells (RBC) aggregation, RBC deformability and whole blood viscosity were also performed. RESULTS: 25(OH)D showed a positive correlation with RBC deformability 0.60 Pa. Subjects with 25(OH)D≤29.00âng/mL had lower RBC deformability 0.60 Pa, and higher RBC aggregation and higher HOMA-IR. Ionized calcium showed a negative correlation with RBC aggregation. Subjects with ionized calcium ≤1.24âmmol/L showed higher RBC aggregation. There was a positive correlation between HOMA-IR and RBC aggregation and HOMA-IR showed a negative correlation with RBC deformability 0.30 Pa. Subjects with HOMA-IR <1.80 showed lower RBC aggregation and higher RBC deformability at 0.30 Pa, 0.60 Pa, 1.20 Pa, 3.0 Pa and 6.0 Pa. CONCLUSION: Low 25(OH)D, low ionized calcium and high HOMA-IR are related to impaired hemorheology in menopause. RBC aggregation and deformability can be used as biomarkers of calcium metabolism and insulin resistance in menopause.
Assuntos
Biomarcadores/sangue , Cálcio/metabolismo , Doenças Cardiovasculares/sangue , Hemorreologia/genética , Resistência à Insulina/genética , Idoso , Deformação Eritrocítica , Feminino , Humanos , Pessoa de Meia-Idade , Pós-MenopausaRESUMO
Aim: Our study aimed to test whether plasma acetylcholinesterase and butyrylcholinesterase enzyme activity were related to the presence and intensity of delirium in acute stroke patients. Methods: We carried out a matched (age and gender) case-control study, in a sample of consecutive patients with an acute infarct or intracerebral haemorrhage (≤7 days). We assessed delirium using the DSM-5 criteria and the Delirium Rating Scale, and we measured plasma acetylcholinesterase and butyrylcholinesterase enzyme activity after the patient's admission in the stroke unit and before hospital discharge. Mantel-Haenszel's chi-square was used to test bivariate associations between cases (delirious patients) and controls (non-delirious patients). Results: At admission in the stroke unit, cases and controls did not present significant differences in plasma acetylcholinesterase or butyrylcholinesterase activity. At hospital discharge (18 cases and 21 controls) patients who have had delirium at admission had higher levels of butyrylcholinesterase activity. Butyrylcholinesterase activity may secondarily increase due to the inflammatory process associated with neuronal dysfunction in delirium patients.
RESUMO
Changes in retinal microcirculation are associated with the development of diabetic retinopathy (DR). However, it is unclear whether such changes also develop in capillary beds of other non-retinal tissues. Here, we investigated microcirculatory changes involving velocity of rolling neutrophils, adherence of neutrophils, and leukostasis during development of retinal vascular lesions in diabetes in other non-retinal tissues. Intravital microscopy was performed on post-capillary venules of cremaster muscle and ear lobe of mice with severe or moderate diabetes and compared to those of non-diabetic mice. Additionally, number and velocity of rolling leukocytes, number of adherent leukocytes, and areas of leukostasis were quantified, and retinal capillary networks were examined for acellular capillaries (AC) and pericyte loss (PL), two prominent vascular lesions characteristic of DR. The number of adherent neutrophils and areas of leukostasis in the cremaster and ear lobe post-capillary venules of diabetic mice was increased compared to those of non-diabetic mice. Similarly, a significant increase in the number of rolling neutrophils and decrease in their rolling velocities compared to those of non-diabetic control mice were observed and severity of diabetes exacerbated these changes. Understanding diabetes-induced microcirculatory changes in cremaster and ear lobe may provide insight into retinal vascular lesion development in DR.
Assuntos
Diabetes Mellitus Experimental/metabolismo , Retinopatia Diabética/metabolismo , Leucócitos/metabolismo , Leucostasia/metabolismo , Microcirculação/fisiologia , Retina/metabolismo , Animais , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patologia , Retinopatia Diabética/genética , Retinopatia Diabética/patologia , Leucócitos/patologia , Leucostasia/genética , Leucostasia/patologia , Masculino , Camundongos , Camundongos Transgênicos , Retina/patologiaRESUMO
BACKGROUND: Myriads of signaling pathways in a single cell function to achieve the highest spatio-temporal integration. Data are accumulating on the role of electromechanical soliton-like waves in signal transduction processes. Theoretical studies strongly suggest feasibility of both classical and quantum computing involving microtubules. AIM: A theoretical study of the role of the complex composed of the plasma membrane and the microtubule-based cytoskeleton as a system that transmits, stores and processes information. METHODS: Theoretical analysis presented here refers to (i) the Penrose-Hameroff theory of consciousness (Orchestrated Objective Reduction; Orch OR), (ii) the description of the centrosome as a reference system for construction of the 3D map of the cell proposed by Regolini, (iii) the Heimburg-Jackson model of the nerve pulse propagation along axons' lipid bilayer as soliton-like electro-mechanical waves. RESULTS AND CONCLUSION: The ideas presented in this paper provide a qualitative model for the decision-making processes in a living cell undergoing a differentiation process. OUTLOOK: This paper paves the way for the real-time live-cell observation of information processing by microtubule-based cytoskeleton and cell fate decision making.
Assuntos
Membrana Celular/metabolismo , Centrossomo/química , Transdução de Sinais , Estresse Mecânico , Actinas/química , Animais , Apoptose , Linhagem da Célula , Citoesqueleto/metabolismo , Tomada de Decisões , Humanos , Íons , Bicamadas Lipídicas/química , Microtúbulos/metabolismoRESUMO
BACKGROUND: Natural and synthetic estrogens seems to have opposite effects on thrombosis and female cardiovascular system, since natural estrogen was supposed to be protective against cardiovascular diseases and synthetic estrogen has been related to thrombosis and cardiovascular diseases. In this work we have investigated if these differences could be related with the effects on those hormones on some hemorheological parameters. OBJECTIVE: The objective of this work was to investigate the hemorheological changes of different concentrations of beta-estradiol and ethinylestradiol, on RBC aggregation and RBC deformability. METHODS: Samples of blood of healthy donors were added with different concentrations of natural beta-estradiol or synthetic ethinylestradiol and were analyzed for red blood cell (RBC) aggregation and RBC deformability. RESULTS: There were no significant changes in RBC aggregation. Both beta-estradiol and ethinylestradiol increase the RBC deformability in shear stresses above 3.0âPa accordingly with the hormone's concentration. CONCLUSIONS: Beta-estradiol and ethinylestradiol enhance RBC deformability dependent of their concentration. These findings may explain the different patterns of thrombotic and cardiovascular effects in different phases of the menstrual cycle or different dosages of oral contraceptive or hormonal replacement therapy.
Assuntos
Deformação Eritrocítica/efeitos dos fármacos , Estradiol/uso terapêutico , Etinilestradiol/uso terapêutico , Hemorreologia , Estradiol/farmacologia , Etinilestradiol/farmacologia , Feminino , HumanosRESUMO
Timolol maleate is a compound used in treatment for reducing increased intra-ocular pressure by limiting aqueous humor production. Decreased erythrocyte deformability (ED), increased activity of erythrocyte acetylcholinesterase (AChE), increased values of nitrosoglutathione (GSNO) and nitic oxide (NO) and decreased plasma levels of NO metabolites, were described in primary open angle glaucoma patients. In healthy human red blood cells (RBCs), timolol is an inhibitor of AChE and induces NO efflux and GSNO efflux from that blood component in lower concentration than those obtained in presence of the natural AChE substrate, acetylcholine (ACh). The signal transduction pathway in RBCs described for NO in dependence of AChE-ACh active complex involves Gi protein, protein tyrosine kinase (PTK like Syk and p53/56Lyn), protein tyrosine phosphatase (PTP) and adenylyl cyclase (AC).The aim of this in vitro study was to verify the effect of timolol maleate in ED, NO efflux and NO derivatives molecules (NOx) like nitrite (NO2-), nitrate (NO3-, peroxynitrite (-ONOO) and GSNO under the presence of PTK, PTP, AC and guanylyl cyclase (GC) enzyme proteins inhibitors.Blood samples from healthy donors were each one divided and were performed aliquots in absence (control aliquots) and presence of timolol or timolol plus each inhibitor and Gi protein uncoupling. No significant differences in erythrocyte NO efflux, GSNO, peroxynitrite, nitrite and nitrate concentrations in response to timolol when compared with the untreated blood samples aliquots were obtained.It was observed an increase in erythrocyte deformability at high shear stresses induced by the simultaneous presence of timolol and band 3 protein dephosphorylation by PTK syk inhibitor. No significant differences where verified in peroxynitrite levels in the blood aliquots in presence of timolol plus each enzyme inhibitor and Gi protein uncoupling in relation to the control aliquots. No variation of GSNO concentration occurs under the presence of timolol and AMGT (PTK lyn inhibitor) besides the significant higher values observed with each one of the other inhibitors. Nitrate concentration increases significantly in all aliquots with timolol plus each one of the inhibitors. The same was observe with nitrite levels with exception of the aliquots with timolol plus AMGT or timolol plus Gi protein uncoupling showing no significant values in relation to the control aliquots.Besides the changes in NO derivative molecules and NO efflux from RBCs obtained in this study with blood samples of healthy donors under the effect of timolol plus each inhibitor of the proteins participants in NO signal transduction mechanism, further analogue studies must be promoted with blood samples of patients with glaucoma or any other inflammatory vascular disease.
Assuntos
Antagonistas Adrenérgicos beta/uso terapêutico , Deformação Eritrocítica/efeitos dos fármacos , Óxido Nítrico/metabolismo , Timolol/uso terapêutico , Antagonistas Adrenérgicos beta/farmacologia , Humanos , Óxido Nítrico/sangue , Timolol/farmacologiaRESUMO
The biochemical properties of erythrocyte or human red blood cell (RBC) membrane acetylcholinesterase (AChE) and its applications on laboratory class and on research are reviewed. Evidence of the biochemical and the pathophysiological properties like the association between the RBC AChE enzyme activity and the clinical and biophysical parameters implicated in several diseases are overviewed, and the achievement of RBC AChE as a biomarker and as a prognostic factor are presented. Beyond its function as an enzyme, a special focus is highlighted in this review for a new function of the RBC AChE, namely a component of the signal transduction pathway of nitric oxide.
Assuntos
Acetilcolinesterase/metabolismo , Esclerose Lateral Amiotrófica/diagnóstico , Hipertensão Essencial/diagnóstico , Glaucoma/diagnóstico , Hemoglobinúria Paroxística/diagnóstico , Doença de Hirschsprung/diagnóstico , Acetilcolina/metabolismo , Esclerose Lateral Amiotrófica/enzimologia , Biomarcadores/metabolismo , Membrana Eritrocítica/enzimologia , Hipertensão Essencial/enzimologia , Feminino , Proteínas Ligadas por GPI/metabolismo , Glaucoma/enzimologia , Hemoglobinúria Paroxística/enzimologia , Doença de Hirschsprung/enzimologia , Humanos , Cinética , Masculino , Óxido Nítrico/metabolismo , Fatores Sexuais , Transdução de SinaisRESUMO
Leukocyte recruitment is an essential stage of the inflammatory response and although the molecular mechanisms of this process are relatively well known, the influence of the hydrodynamic effects that govern the inflammatory response are still under study. In this paper we made use of the images and experimental parameters obtained by intravital microscopy in an in vivo animal model of inflammation to track the leukocytes trajectories and measure their velocities and diameters. Using a recent validated mathematical model describing the coupled deformation-flow of an individual leukocyte in a microchannel, numerical simulations of an individual and of two leukocytes under flow were performed. The results showed that velocity plays an important role in the motion, deformation and attraction of the cells during an inflammatory response. In fact, for higher inlet velocities the cell movement along the endothelial wall is accelerated and the attraction forces break faster. These results highlight the role of the mechanical properties of the blood, namely the ones influenced by the velocity field, in the case of inflammation.
Assuntos
Endotélio Vascular/fisiopatologia , Hemorreologia , Inflamação/fisiopatologia , Migração e Rolagem de Leucócitos , Leucócitos/imunologia , Animais , Velocidade do Fluxo Sanguíneo , Simulação por Computador , Modelos Animais de Doenças , Endotélio Vascular/imunologia , Análise de Elementos Finitos , Hidrodinâmica , Inflamação/sangue , Inflamação/imunologia , Microscopia Intravital , Masculino , Camundongos Transgênicos , Modelos Cardiovasculares , Análise Numérica Assistida por Computador , Fluxo Sanguíneo Regional , Fatores de TempoRESUMO
BACKGROUND: Glaucoma is an optic neuropathy associated with vascular dysregulation and increased intra-ocular pressure (IOP). Timolol is used as treatment for reducing IOP, by limiting aqueous humour production. Increased NOS expression as well as decreased levels of nitric oxide (NO) metabolites, and high activity of erythrocyte acetylcholinesterase (AChE) were observed in primary open angle glaucoma patients. OBJECTIVE: This ex vivo study aims to evaluate timolol effect in NO efflux and its derivatives in glaucoma patient's erythrocytes. METHODS: Venous blood from 15 glaucoma patients was collected. Erythrocyte suspensions were incubated with the AChE modulators acetylcholine (ACh) and timolol at 10 µM. Erythrocyte NO efflux and S-nitrosoglutathione (GSNO) concentration were measured. RESULTS: No significant differences were obtained in erythrocyte NO efflux and GSNO concentration in response to ACh or timolol when compared with the untreated erythrocytes of glaucoma patients. When comparing the same incubation conditions for erythrocyte suspensions between glaucoma patients and healthy subjects, those from glaucoma patients showed higher NO efflux in presence and absence of timolol, and higher values of GSNO in the presence of timolol. CONCLUSIONS: We demonstrated that erythrocytes from glaucoma patients have similar availability to release NO both in absence and presence of timolol, and have higher GSNO values in presence of timolol.
Assuntos
Antagonistas Adrenérgicos beta/uso terapêutico , Eritrócitos/metabolismo , Glaucoma/metabolismo , Óxido Nítrico/sangue , Timolol/uso terapêutico , Antagonistas Adrenérgicos beta/administração & dosagem , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Timolol/administração & dosagemRESUMO
INTRODUCTION: Acetylcholinesterase (AChE) is located on outer surface of erythrocyte membrane. Gender-related differences in erythrocyte AChE enzyme activity had been verified in young adults. It is also known that binding of acetylcholine (ACh) with AChE on erythrocyte membrane initiates a signal transduction mechanism that stimulates nitric oxide (NO) efflux. AIMS: This ex vivo study was done to compare the amount of NO efflux obtained from erythrocytes of healthy donors in males and females. METHODS: We included 66 gender age-matched healthy donors (40-60 years old). We performed quantification of erythrocyte NO efflux from erythrocytes and of the membrane AChE enzyme activity. RESULTS: There are no significant differences in NO efflux from erythrocytes between men and women. Regarding AChE enzyme activity values, in this range of age, no differences between genders were obtained. However, the values of AChE enzyme activity in the third quartile of NO efflux values were significantly higher (pâ<â0.05) in women than in men. CONCLUSIONS: The efflux of NO from erythrocyte of healthy humans did not change with gender. For the same range of values of NO efflux from erythrocytes, in both gender, it was verified higher values of AChE enzyme activity in women.
Assuntos
Acetilcolinesterase/metabolismo , Eritrócitos/metabolismo , Óxido Nítrico/sangue , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
AIMS: Oxidized low density lipoprotein (ox-LDL) has been reported as an inhibitor of nitric oxide (NO)-mediated dilatation in microcirculation. Oxidized LDL effect on NO metabolism of erythrocytes is not known. Therefore, this study aims to evaluate the effect of ox-LDL on erythrocytes NO metabolism. METHODS: The effect of different concentrations of human purified ox-LDL (25, 50 and 100 µg/mL) on NO metabolism was evaluated on blood of healthy subjects. RESULTS: An inhibitory effect of higher concentrations of ox-LDL on erythrocyte NO efflux levels was verified. Concentrations of NO efflux from erythrocytes were lower as consequence of treatments with 50 µg/mL ox-LDL treatment (1.6±0.27 nM) and 100 µg/mL ox-LDL treatment (1.3±0.22 nM) than control (1.9±0.28 nM). Opposite, ox-LDL incubation has a positive effect on GSNO content of erythrocytes. That effect is proportional to concentrations of ox-LDL treatments (10.8±1.4 nM for 25 µg/mL, 12.9±1.5 nM for 50 µg/mL and 12.1±1.9 nM for 100 µg/mL) and is significant relative to control (8.56±0.76 µM) and ACh (8.9±0.52 µM) aliquots. CONCLUSIONS: Presence of oxidized LDL in erythrocyte NO metabolism induces a decrease of NO efflux amount and an increase on intra-erythrocyte GSNO concentrations. These results suggest a role of ox-LDL in mobilization of NO between NO derivatives molecules in dependence of oxidized LDL concentration. An anti - reactive nitrogene role can be attributed to ox-LDL for its contribution in the erythrocyte scavenged ability for nitric oxide.
Assuntos
Eritrócitos/efeitos dos fármacos , Lipoproteínas LDL/metabolismo , Óxido Nítrico/metabolismo , Humanos , Óxido Nítrico/sangueRESUMO
AIM: Soluble CD40 ligand (sCD40L) has been considered as a marker of thrombosis and inflammation in several diseases, including sepsis. Recent studies challenge this view and point to a role of sCD40L in vascular and endothelial function. An indication of that association in sepsis has not been obtained so far. Therefore, herein we evaluated association between sCD40L and markers of hemorheology and inflammation on context of septic shock. METHODS: Time-changes of sCD40L levels over 72 hours of Intensive Care Unit (ICU) internment were assessed in 22 patients with septic shock and compared with 36 healthy volunteers. Association of sCD40L levels with erythrocyte deformability and aggregation (as markers of hemorheology), plasma concentrations of haemoglobin (Hb, as markers of endothelial function) and white blood cells (WBC) count (as marker of low-grade inflammation) were assessed in patients with septic shock. RESULTS: At ICU admission, sCD40L concentrations in patients with septic shock were lower (pâ=â0.024) than levels of healthy volunteers. However, sCD40L did not change over 72 hours of internment (Fâ=â2.1, pâ=â0.137). Soluble CD40L levels in patients with septic shock at ICU admission correlate with concentrations of Hb (râ=â0.61, pâ=â0.00) and WBC count (râ=â0.63, pâ=â0.00), but not to erythrocyte deformability (r≥0.157, p≤0.235) and aggregation (r≥-0.109, p≤0.192). CONCLUSIONS: These results seem to highlight a possible association of sCD40L to endothelial function and inflammation in septic shock context.
Assuntos
Biomarcadores/sangue , Ligante de CD40/metabolismo , Choque Séptico/genética , Choque Séptico/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Hemorreologia , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Nitric oxide (NO) produced by endothelial cells interacts with erythrocyte through band 3 protein, being scavenged by haemoglobin. A signal transduction mechanism involving protein Gi and protein band 3 stimulates erythrocyte NO efflux when acetylcholine (ACh) binds to erythrocyte membrane acetylcholinesterase. Binding of normal plasma fibrinogen (Fib) levels, to erythrocyte membrane CD47 decreases the NO efflux. When high Fib concentration and ACh were present the efflux of NO from erythrocytes was normalized. The increased NO efflux from erythrocytes in presence of high Fib concentration and band 3 phosphorylation is reinforced in the presence of 4N1K an agonist peptide of CD47. When both Fib and 4N1K are present the NO efflux from erythrocytes is higher or not affected according lower or high levels of cAMP. Erythrocyte NO efflux in patients with systemic lupus erythematous and rheumatoid attrite was significantly negative associated with carotid intima-media thickness. In patients with amyotrophic lateral sclerosis erythrocyte NO content is preserved and an inverse association between respiratory function and NO efflux from the erythrocyte was verified. Sepsis patients before dead at 24âh showed higher efflux of NO from erythrocytes that worsening the blood sub lingual microcirculation observed by high unequal blood flow and high microvascular flow index. The in vivo animal models either of inflammation or of hypertension evidenced that the NO efflux from erythrocyte decrease as a compensatory mechanism. All studies conducted since 2000 where we demonstrated the existence NO inside the erythrocyte by fluorescence microscopy, and after their signaling pathway needs more development translational research for news therapeutics and further application in not invasive therapy to vascular inflammatory diseases.
Assuntos
Eritrócitos/fisiologia , Fibrinogênio/metabolismo , Óxido Nítrico/fisiologia , Animais , Eritrócitos/metabolismo , Humanos , Óxido Nítrico/sangue , Transdução de SinaisRESUMO
INTRODUCTION: Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor system. It has been hypothesised that red blood cells (RBCs) may be involved in the disease process by the release of damaging molecules. OBJECTIVE: The aim of this ex vivo study is to compare RBCs biochemical and hemorheological parameters between ALS patients and healthy donors to identify novel biomarkers of the ALS disease. METHODS: We included 82 ALS patients and 40 gender age-matched healthy donors. We performed quantification of erythrocyte aggregation and deformability, nitric oxide (NO) efflux from RBCs, acetylcholinesterase (AChE) enzyme activity and intraerythrocytic concentration of nitrite, nitrate and S-nitrosogluthatione (GSNO). RESULTS: Erythrocyte deformability and AChE activity were increased in patients with ALS in comparison to healthy donors. NO efflux from RBCs and concentration of intraerythrocytic nitrite were lower in ALS patients. In patients, we found that for higher NO range of values the respiratory function is worse and that for higher AChE range of values the RBCs nitrite content increase. CONCLUSION: The results of the present study indicate that NO efflux from RBCs and RBCs AChE should be further explored as potential biomarkers for ALS.
Assuntos
Esclerose Lateral Amiotrófica/sangue , Biomarcadores/sangue , Eritrócitos/citologia , Eritrócitos/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Agregação Eritrocítica , Deformação Eritrocítica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/sangue , Nitritos/sangueRESUMO
Involvement of soluble CD40 ligand (sCD40L) in thrombosis and inflammation on the context of coronary artery disease is currently being revised. In that perspective, we had studied the association of sCD40L with markers of platelet activation and markers of endothelial and vascular function. On that cohort, a stratification of patients with acute myocardial infarction (AMI) 1 month after percutaneous coronary intervention (PCI) was observed based on concentrations of sCD40L. The study intended to identify the groups of AMI patients with different profiles of sCD40L concentrations and verify how medication, clinical evolution, biochemical data, and markers of regulation of endothelial function at genetic (endothelial nitric oxide synthase polymorphisms) and post-transcriptional levels (circulating microRNAs) affect sCD40L serum levels. Lower quartiles of sCD40L (<2.3 ng/mL) were associated with higher concentrations of N-terminal pro-brain natriuretic peptide (NT-proBNP), high frequency of G894T polymorphism, and altered expression of a set of microRNAs assumed to be involved in the regulation of endothelial and cardiac function and myocardium hypertrophy, relative to patients in sCD40L upper quartiles. A characteristic sCD40L variation pattern in STEMI patients was identified. Low levels of sCD40L 1 month after PCI distinguish STEMI patients with worse prognosis, a compromised cardiac healing, and a persistent endothelial dysfunction, as given by the association between sCD40L, NT-proBNP, G894T polymorphism, and specific profile of miRNA expression. These results suggest sCD40L could have a prognostic value in STEMI patients.
Assuntos
Ligante de CD40/sangue , Eletrocardiografia , Infarto do Miocárdio/sangue , Infarto do Miocárdio/diagnóstico por imagem , Idoso , Estudos de Casos e Controles , Feminino , Perfilação da Expressão Gênica , Genótipo , Humanos , Estudos Longitudinais , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Infarto do Miocárdio/genética , Peptídeo Natriurético Encefálico/sangue , Óxido Nítrico Sintase Tipo III/genética , Fragmentos de Peptídeos/sangue , Intervenção Coronária Percutânea , SolubilidadeRESUMO
We aim to establish an in vivo animal model of acute inflammation using PAF (platelet activating factor) as inflammatory agent and to study the erythrocyte deformability changes induced by the inflammatory response. Counting the number of rolling and adherent neutrophils to endothelium after 2, 4 and 6h of intrascrotal injection of PAF we showed the induction of an inflammatory state. Blood samples are collected in order to measure the erythrocyte deformability and to quantify NO efflux from the red blood cells (RBCs). The results show an increased number of rolling and adherent neutrophils after 2h and 4h of inflammation as well as decreased values of erythrocyte deformability in the same time-points. This result is in line with the need of a low blood viscosity to the recruitment process that will improve leukocyte migration towards the endothelial wall. NO efflux from RBCs is also affected by the inflammatory response at the first hours of inflammation. This animal model demonstrates in vivo the association between an acute inflammatory response and the rheological properties of the blood, namely the RBCs deformability. For those reasons we consider this as an adequate model to study acute inflammatory responses as well as hemorheological parameters.