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1.
J Thromb Thrombolysis ; 48(2): 187-194, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31177487

RESUMO

Relatively little information is known about the definitive role of phosphatidylserine (PS) in the hypercoagulability of heart failure (HF). Our objectives were to assess the levels of PS exposure on microparticles (MPs) and blood cells (BCs) in each group of HF patients and to evaluate their procoagulant activity (PCA). HF patients in each NYHA functional class II-IV (II n = 30, III n = 30, IV n = 30) and healthy controls (n = 25) were enrolled in the present study. PS exposure on MPs, BCs was analyzed with flow cytometry. MPs were classified based on their cellular origin: platelets (CD41a+), neutrophils (CD66b+), endothelial cells (CD31+CD41a-), erythrocytes (CD235a+), monocytes (CD14+), T lymphocytes (CD3+), and B lymphocytes (CD19+). PCA was evaluated by clotting time, extrinsic/intrinsic FXa and prothrombinase production assays, as well as fibrin formation assays. Inhibition assays of PCA of PS+ BCs and MPs were performed by lactadherin. There was no significant difference in MP cellular origin between healthy and HF subjects. However, the total number of PS+ MPs was significantly increased in HF patients compared with healthy controls. In addition, circulating PS+ BCs cooperated with PS+ MPs to markedly shorten coagulation time and dramatically increase FXa/thrombin generation and fibrin formation in each HF group. Moreover, blockade of exposed PS on BCs and MPs with lactadherin inhibited PCA by approximately 80%. Our results lead us to believe that exposing PS on the injured BCs and MPs played a pivotal role in the hypercoagulability state in HF patients.


Assuntos
Micropartículas Derivadas de Células/fisiologia , Células Endoteliais/fisiologia , Insuficiência Cardíaca/sangue , Fosfatidilserinas/metabolismo , Trombofilia/etiologia , Adulto , Idoso , Células Sanguíneas/patologia , Coagulação Sanguínea , Testes de Coagulação Sanguínea , Estudos de Casos e Controles , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade
2.
Adv Clin Exp Med ; 28(5): 683-692, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30712335

RESUMO

BACKGROUND: Chronic and oxidative stress promotes injury to the endothelium. This happens early in the disease and novel biomarkers describing the rate of the damage may be important in early diagnostics and prevention. Microvesicles are shed from endothelial cells in response to oxidative stress, inflammation, coagulation, and angiogenesis. Their increased level in plasma could reflect the state of the endothelium. OBJECTIVES: The objective of this study was to test the association between oxidative and chronic stress markers, atherosclerosis risk factors and endothelial microvesicle (EMV) count in peripheral blood. MATERIAL AND METHODS: The study included 81 males, aged 25-55 years and apparently healthy. Venous blood samples were labeled with anti-CD144-FITC, anti-CD105-BV421, anti-CD42a-PerCP, anti-CD62e-PE, anti-CD31-APCy7, and anti-CD61-APC (BD Biosciences, San Jose, USA), and tested using a BD LSR Fortessa cytometer (BD Biosciences). Events were gated on forward and side-scattered light parameters. Malondialdehyde (MDA) and cortisol concentrations were measured using high-performance liquid chromatography (HPLC). RESULTS: Four populations of EMV expressing a combination of CD105+, CD31+, CD144+, and CD62e with CD42aor CD42a+ markers were examined. We found correlations between MDA concentration and hair cortisol and a total count of CD144+ microvesicles, and weak correlations with diastolic blood pressure (DBP) (p = 0.003, r = 0.324) and systolic blood pressure (SBP) (p = 0.016, r = 0.267), especially with the microvesicles carrying CD62e. There was a median difference of CD105+ microvesicle count between smoking (n = 13) and non-smoking (n = 68) individuals. A predictive model showed an association between CD144+ microvesicle counts with cortisol and MDA concentrations and waist circumference. CONCLUSIONS: In conclusion, our data and predictive model showed that the total counts of microvesicle populations were associated with stress-related parameters - cortisol and MDA concentrations; expression of CD62e in various populations of EMV and the ratio of CD144+ to CD105+/CD62e+ were associated with increased DBP and SBP, and also with total cholesterol concentration in healthy young male population.


Assuntos
Movimento Celular/fisiologia , Micropartículas Derivadas de Células/fisiologia , Células Endoteliais/fisiologia , Estresse Oxidativo , Adulto , Biomarcadores , Endotélio , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade
3.
Semin Liver Dis ; 39(1): 70-77, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30654391

RESUMO

Extracellular vesicles, comprising exosomes, microvesicles, and apoptotic bodies, represent an emerging field in disease diagnostics and prognosis. They can be isolated from peripheral blood of patients as well as from other body fluids and can therefore be considered a minimally invasive liquid biopsy screening tool. Especially their surface antigen composition can reveal information about disease backgrounds. For several liver diseases, including fatal hepatocellular and cholangiocellular carcinoma as well as other nonmalignant liver disorders such as nonalcoholic fatty liver disease, alcoholic hepatitis, or acute liver failure, it has been shown that extracellular vesicle (EV) surface profiling can be useful for disease diagnosis and prognosis. This review focuses on latest advances in these areas to improve liver disorder detection and management. Additionally, the authors will discuss possible therapeutic applications of EVs in liver diseases, which might be a potent treatment option in the future.


Assuntos
Micropartículas Derivadas de Células/fisiologia , Exossomos/fisiologia , Hepatopatias/diagnóstico , Animais , Biomarcadores/sangue , Humanos , Hepatopatias/terapia , Camundongos
4.
Hum Cell ; 32(1): 64-74, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30506278

RESUMO

Studies have demonstrated that mesenchymal stem cells (MSCs) can promote tumor growth, and MSC microvesicles (MVs) are very important in the tumor microenvironment and information transfer between cells during tumorigenesis and development. However, the potential effects and mechanisms of MSC-MVs on tumor growth are still controversial. Here in this study, we investigated the roles and effects of human bone marrow MSC-MVs (hBMSC-MVs) on human osteosarcoma (U2OS) cell growth under hypoxia in vitro and in vivo. BMSC-MVs were harvested and purified by ultracentrifugation. U2OS cells were treated with different concentrations of hBMSC-MVs under hypoxia. Cell viability and migration was measured by MTT test, transwell invasion assay and scratch migration assay. The expression of the signaling molecules of AKT, VEGF, GLUT1 and Bax, cleaved-caspase3 in U2OS cells cultured with MVs under hypoxia was determined by western blot. U2OS/siHIF-1α or U2OS/NC cells mixed with/without MVs were subcutaneously injected into nude mice; the tumor size and weight were detected. We found that hBMSC-MVs promoted U2OS cell proliferation and migration under hypoxia in vitro, and that was partially associated with the PI3K/AKT and HIF-1α pathways. MVs co-injected with U2OS cells promoted tumor growth in a mouse xenograft model. siHIF-1α transfection reversed these changes to some extent. The function of hBMSC-MVs on U2OS cell progression and tumor growth was associated with PI3K/AKT and HIF-1α pathway under hypoxia. These findings support a new mechanism suggesting the contribution of MSC-MVs to tumor growth.


Assuntos
Células da Medula Óssea/citologia , Hipóxia Celular/fisiologia , Micropartículas Derivadas de Células/fisiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/fisiologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/fisiologia , Osteossarcoma/genética , Osteossarcoma/patologia , Fosfatidilinositol 3-Quinases/fisiologia , Proteínas Proto-Oncogênicas c-akt/fisiologia , Transdução de Sinais/fisiologia , Animais , Movimento Celular/genética , Proliferação de Células/genética , Sobrevivência Celular/genética , Humanos , Camundongos Nus , Invasividade Neoplásica/genética , Células Tumorais Cultivadas
5.
Blood Cells Mol Dis ; 74: 37-43, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30454964

RESUMO

BACKGROUND AND AIMS: Myelodysplasia (MDS) is characterised by abnormal haematopoiesis and increased risk of bleeding. Microvesicles (MV) play a key role in coagulation and their impact in MDS is unknown. METHODS: Platelet free plasma from 35 red-cell transfusion-dependent MDS patients and 15 controls were analysed. Pro-coagulant function was assessed by the XaCT assay and by thrombin generation (ETP). Total MV were enumerated by nano-tracking analysis. MV subsets were quantified by flow cytometry after staining with specific antibodies for various endovascular cell types. Small RNA was quantitated and sequenced. The MV measurements were correlated with MDS clinical risk scores and level of transfusion dependence. RESULTS: The pro-coagulant function of MV was significantly lower in MDS. All the MV subtypes, as measured by flow cytometric markers, were also significantly lower. The small RNA and miRNA cargo were significantly higher in MDS. The miRNA profile showed that mir-28 and mir-LETD7 were under expressed whilst mir-584J and mir-4485 were over expressed in MV from MDS. CONCLUSIONS: Circulating MV in MDS show reduced pro-coagulant functional activity, reduced subtypes by flow cytometry and significantly different miRNA content. However, the levels or subtypes of MV did not predict the clinical phenotype or level of transfusion dependence.


Assuntos
Micropartículas Derivadas de Células/fisiologia , MicroRNAs/análise , Síndromes Mielodisplásicas/patologia , Trombofilia/etiologia , Testes de Coagulação Sanguínea , Estudos de Casos e Controles , Micropartículas Derivadas de Células/genética , Citometria de Fluxo , Humanos
6.
Cell Immunol ; 336: 1-11, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30538031

RESUMO

Patients with rheumatoid arthritis (RA) have increased amount of platelet-derived microparticles (PMPs) positive for citrullinated peptides (CPs) that form immune complexes (PMPs-ICs). Monocytes are important inflammatory mediators that play a role in the clearance of PMPs-ICs. We aimed to generate PMPs-ICs in vitro and determine its effect on monocytes from patients with RA and healthy individuals (HI). PMPs from patients showed platelet markers, mitochondria content, and phosphatidylserine exposure similar to PMPs from HI. However, patients had a higher frequency of IgG+ and CPs+ vesicles than HI. PMPs-ICs generated in vitro were similar to the circulating vesicles of patients with respect to IgG- and CPs-positivity. PMPs-ICs induced pro-inflammatory cytokines and CX3CR1 expression in monocytes from HI, and IL-10 and CD36 upregulation in monocytes from patients. These results suggest that PMPs-ICs induce activation of monocytes, with a pro-inflammatory response in HI and a more tolerant response in cells of patients with RA.


Assuntos
Artrite Reumatoide/imunologia , Plaquetas/fisiologia , Micropartículas Derivadas de Células/fisiologia , Monócitos/fisiologia , Adulto , Idoso , Complexo Antígeno-Anticorpo/imunologia , Receptor 1 de Quimiocina CX3C/análise , Citrulinação , Citocinas/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
7.
J Hypertens ; 37(1): 154-166, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30063637

RESUMO

OBJECTIVES: To assess the acute effects of nicotine-containing electronic cigarettes versus tobacco smoking on vascular and respiratory function and circulating microparticles, particularly platelet microparticles (PMPs, biomarker of haemostasis/thrombosis) and endothelial microparticles (EMPs, biomarker of endothelial function). METHODS: Heart rate (HR), blood pressure, reactive hyperaemia index (RHI, microvascular reactivity), augmentation index (arterial stiffness) and respiratory function were assessed in 20 smokers immediately before and after electronic cigarettes use and tobacco smoking. The number of microparticles was determined by flow cytometry using counting beads as a reference. Labelling with Annexin-V was used to detect the total microparticle fraction. EMPs were characterized as CD31+CD42- and PMPs as CD31+CD42+. RESULTS: HR increased after electronic cigarettes use and tobacco smoking (P < 0.001), whereas blood pressure remained unchanged (P > 0.05). RHI (P = 0.006), augmentation index (P = 0.010) but not augmentation index standardized to HR 75 bpm (P > 0.05) increased with electronic cigarettes use but not with tobacco smoking. Following tobacco smoking, there was a significant increase in total microparticles (P < 0.001), EMPs (P < 0.001) and PMPs (P < 0.001). In contrast, electronic cigarettes were only associated with an increase in PMPs (P < 0.001), with no significant changes in the total microparticle fraction or EMPs (all P > 0.05). Peak expiratory flow significantly decreased following electronic cigarettes use (P = 0.019). CONCLUSION: Our results demonstrate that acute exposure to tobacco smoking as well as electronic cigarettes influences vascular and respiratory function. Where tobacco smoking significantly increased microparticle formation, indicative of possible endothelial injury, electronic cigarettes use induced vasoreactivity and decreased peak expiratory flow. These findings suggest that both electronic cigarettes and tobacco smoking negatively impact vascular function.


Assuntos
Pressão Sanguínea/fisiologia , Fumar Cigarros , Frequência Cardíaca/fisiologia , Vaping , Micropartículas Derivadas de Células/fisiologia , Fumar Cigarros/sangue , Fumar Cigarros/fisiopatologia , Estudos Cross-Over , Humanos , Respiração , Vaping/sangue , Vaping/fisiopatologia
8.
Curr Stem Cell Res Ther ; 14(1): 9-13, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30152289

RESUMO

Cardiovascular Disease (CVD) is one of the world-wide healthcare problem that involves the heart or blood vessels. CVD includes myocardial infarction and coronary artery diseases (CAD). Dysfunctional myocardial cells are leading causes of low cardiac output or ventricular dysfunction after cardiac arrest and may contribute to the progression of CVD which could not generate new cardiomyocytes in human adult heart. The mesenchymal stem cells (MSCs) which are present in adult marrow can self-renew and have the capacity of differentiation into multiple types of cells including cardiomyocytes. Recent biochemical analyses greatly revealed that several regulators of MSCs, such as HGF, PDGF, Wnt, and Notch-1 signaling pathways have been shown to be involved in the proliferation and differentiation into cardiomyocytes. Preclinical studies are paving the way for further applications of MSCs in the repair of myocardial infarction. In this study, we discuss and summarize the paracrine mechanisms involved in MSCs differentiation into cardiomyocytes.


Assuntos
Diferenciação Celular , Células-Tronco Mesenquimais/fisiologia , Miócitos Cardíacos/fisiologia , Comunicação Parácrina/fisiologia , Antígenos de Superfície/biossíntese , Proliferação de Células , Micropartículas Derivadas de Células/fisiologia , Doença da Artéria Coronariana/terapia , Citocinas/fisiologia , Exossomos/fisiologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Transplante de Células-Tronco Mesenquimais/psicologia , Células-Tronco Mesenquimais/citologia , Infarto do Miocárdio/terapia , Miócitos Cardíacos/citologia , Proteínas Proto-Oncogênicas c-met/fisiologia , Receptores do Fator de Crescimento Derivado de Plaquetas/fisiologia , Via de Sinalização Wnt/fisiologia
9.
Oncol Rep ; 40(6): 3335-3345, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30272301

RESUMO

Leukocyte­derived microparticles (LMPs) include neutrophil­, lymphocyte­ and monocyte­derived MPs. LMPs act as proinflammatory mediators in autoimmune diseases, infectious diseases and vascular diseases. The present study examined the hypothesis that the percentage of LMPs was increased in patients with inflamed odontogenic keratocysts (OKCs), and investigated the biological effects of Jurkat cell­derived MPs on the fibroblasts of OKCs in vitro. Cyst fluid MPs, obtained by centrifugation of samples from 20 patients with inflamed OKCs, 3 patients with uninflamed OKCs, 15 patients with radicular cysts (RCs) and 12 patients with inflamed dentigerous cysts (DCs), were analyzed by transmission electron microscopy, dynamic light scattering and immunofluorescence staining. The percentages and concentrations of cyst fluid LMPs were further determined by flow cytometry. The cytokine levels of apoptotic Jurkat cell­derived MPs and Jurkat cell supernatants were compared by cytokine antibody arrays. Fibroblasts were isolated from 3 patients with OKC and co­cultured with apoptotic Jurkat cell­derived MPs with or without interleukin (IL)­15Rα to detect the levels of matrix metallopeptidase 9 (MMP­9) and receptor activator of nuclear factor­κB ligand (RANKL) by reverse transcription­quantitative polymerase chain reaction and enzyme­linked immunosorbent assay. The supernatant from Jurkat MPs­treated fibroblasts was collected to make conditioned medium in which the osteoclastogenesis of Raw264.7 cells was determined. Antibodies against human soluble (s)RANKL were added to the conditioned medium to investigate the inhibitory effects. Mean percentages of lymphocyte­ and neutrophil­derived MPs were significantly higher in inflamed OKCs than in DCs. Significant elevations in IL­15 were detected in apoptotic Jurkat cell­derived MPs compared with that in Jurkat cell supernatant. Furthermore, higher levels of MMP­9 and RANKL were detected in Jurkat cell MP­treated OKC fibroblasts, and this was partially blocked by IL­15Rα. Increased osteoclast­like cell formation was observed in the Jurkat MPs­treated fibroblast supernatant and Raw264.7 co­culture groups. The anti­human sRANKL antibody in the Jurkat MPs­treated fibroblast supernatant group decreased the osteoclastogenesis of the Raw264.7 cells. These results indicate that LMPs serve as novel communication tools that contribute toward the bone resorption of inflamed OKCs by inducing RANKL of OKC fibroblasts via IL­15.


Assuntos
Micropartículas Derivadas de Células/fisiologia , Interleucina-15/metabolismo , Linfócitos/citologia , Cistos Odontogênicos/imunologia , Ligante RANK/metabolismo , Adolescente , Adulto , Idoso , Animais , Micropartículas Derivadas de Células/metabolismo , Células Cultivadas , Criança , Técnicas de Cocultura , Feminino , Humanos , Células Jurkat , Linfócitos/metabolismo , Masculino , Camundongos , Pessoa de Meia-Idade , Células RAW 264.7 , Adulto Jovem
10.
J Am Soc Nephrol ; 29(11): 2671-2695, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30341150

RESUMO

BACKGROUND: Glomerular endothelium dysfunction, which plays a crucial role in the pathogenesis of early diabetic nephropathy, might be caused by circulating metabolic abnormalities. Platelet microparticles, extracellular vesicles released from activated platelets, have recently emerged as a novel regulator of vascular dysfunction. METHODS: We studied the effects of platelet microparticles on glomerular endothelial injury in early diabetic nephropathy in rats with streptozotocin-induced diabetes and primary rat glomerular endothelial cells. Isolated platelet microparticles were measured by flow cytometry. RESULTS: Plasma platelet microparticles were significantly increased in diabetic rats, an effect inhibited in aspirin-treated animals. In cultured glomerular endothelial cells, platelet microparticles induced production of reactive oxygen species, decreased nitric oxide levels, inhibited activities of endothelial nitric oxide synthase and SOD, increased permeability of the glomerular endothelium barrier, and reduced thickness of the endothelial surface layer. Conversely, inhibition of platelet microparticles in vivo by aspirin improved glomerular endothelial injury. Further analysis showed that platelet microparticles activated the mammalian target of rapamycin complex 1 (mTORC1) pathway in glomerular endothelial cells; inhibition of the mTORC1 pathway by rapamycin or raptor siRNA significantly protected against microparticle-induced glomerular endothelial injury in vivo and in vitro. Moreover, platelet microparticle-derived chemokine ligand 7 (CXCL7) contributed to glomerular endothelial injury, and antagonizing CXCL7 using CXCL7-neutralizing antibody or blocking CXCL7 receptors with a competitive inhibitor of CXCR1 and CXCR2 dramatically attenuated such injury. CONCLUSIONS: These findings demonstrate a pathogenic role of platelet microparticles in glomerular endothelium dysfunction, and suggest a potential therapeutic target, CXCL7, for treatment of early diabetic nephropathy.


Assuntos
Plaquetas/fisiologia , Micropartículas Derivadas de Células/fisiologia , Diabetes Mellitus Experimental/sangue , Nefropatias Diabéticas/sangue , Glomérulos Renais/patologia , Animais , Aspirina/farmacologia , Plaquetas/efeitos dos fármacos , Plaquetas/patologia , Micropartículas Derivadas de Células/efeitos dos fármacos , Micropartículas Derivadas de Células/patologia , Células Cultivadas , Quimiocinas CXC/fisiologia , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Experimental/fisiopatologia , Nefropatias Diabéticas/patologia , Nefropatias Diabéticas/fisiopatologia , Células Endoteliais/patologia , Glomérulos Renais/irrigação sanguínea , Glomérulos Renais/efeitos dos fármacos , Masculino , Alvo Mecanístico do Complexo 1 de Rapamicina/antagonistas & inibidores , Alvo Mecanístico do Complexo 1 de Rapamicina/fisiologia , Ativação Plaquetária , Ratos , Ratos Sprague-Dawley , Transdução de Sinais
11.
Nat Commun ; 9(1): 3630, 2018 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-30194420

RESUMO

Microvilli on T cells have been proposed to survey surfaces of antigen-presenting cells (APC) or facilitate adhesion under flow; however, whether they serve essential functions during T cell activation remains unclear. Here we show that antigen-specific T cells deposit membrane particles derived from microvilli onto the surface of cognate antigen-bearing APCs. Microvilli carry T cell receptors (TCR) at all stages of T cell activation and are released as large TCR-enriched, T cell microvilli particles (TMP) in a process of trogocytosis. These microvilli exclusively contain protein arrestin-domain-containing protein 1, which is directly involved in membrane budding and, in combination with vacuolar protein-sorting-associated protein 4, transforms large TMPs into smaller, exosome-sized TMPs. Notably, TMPs from CD4+ T cells are enriched with LFA-2/CD2 and various cytokines involved in activating dendritic cells. Collectively, these results demonstrate that T cell microvilli constitute "immunological synaptosomes" that carry T cell messages to APCs.


Assuntos
Linfócitos T CD4-Positivos/fisiologia , Microvilosidades/fisiologia , Animais , Células Apresentadoras de Antígenos , Linfócitos T CD4-Positivos/ultraestrutura , Micropartículas Derivadas de Células/fisiologia , Células Dendríticas/fisiologia , Células HEK293 , Humanos , Células Jurkat , Camundongos , Receptores de Antígenos de Linfócitos T/metabolismo , Sinaptossomos
12.
PLoS One ; 13(9): e0203991, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30231080

RESUMO

Megakaryocytes (Mks) derive from hematopoietic stem and progenitor cells (HSPCs) in the bone marrow and develop into large, polyploid cells that eventually give rise to platelets. As Mks mature, they migrate from the bone marrow niche into the vasculature, where they are exposed to shear forces from blood flow, releasing Mk particles (platelet-like particles (PLPs), pro/preplatelets (PPTs), and Mk microparticles (MkMPs)) into circulation. We have previously shown that transcription factor p53 is important in Mk maturation, and that physiological levels of shear promote Mk particle generation and platelet biogenesis. Here we examine the role of p53 in the Mk shear-stress response. We show that p53 is acetylated in response to shear in both immature and mature Mks, and that decreased expression of deacetylase HDAC1, and increased expression of the acetyltransferases p300 and PCAF might be responsible for these changes. We also examined the hypothesis that p53 might be involved in the shear-induced Caspase 3 activation, phosphatidylserine (PS) externalization, and increased biogenesis of PLPs, PPTs, and MkMPs. We show that p53 is involved in all these shear-induced processes. We show that in response to shear, acetyl-p53 binds Bax, cytochrome c is released from mitochondria, and Caspase 9 is activated. We also show that shear-stimulated Caspase 9 activation and Mk particle biogenesis depend on transcription-independent p53-induced apoptosis (TIPA), but PS externalization is not. This is the first report to show that shear flow stimulates TIPA and that Caspase 9 activation and Mk-particle biogenesis are directly modulated by TIPA.


Assuntos
Apoptose/fisiologia , Megacariócitos/citologia , Megacariócitos/metabolismo , Trombopoese/fisiologia , Proteína Supressora de Tumor p53/metabolismo , Acetilação , Antígenos CD34/metabolismo , Apoptose/genética , Plaquetas/citologia , Plaquetas/metabolismo , Caspase 3/metabolismo , Caspase 9/metabolismo , Diferenciação Celular , Linhagem Celular , Micropartículas Derivadas de Células/fisiologia , Citocromos c/metabolismo , Ativação Enzimática , Técnicas de Silenciamento de Genes , Humanos , Mitocôndrias/metabolismo , Modelos Biológicos , Fosfatidilserinas/metabolismo , Estresse Mecânico , Trombopoese/genética , Transcrição Genética , Proteína Supressora de Tumor p53/antagonistas & inibidores , Proteína Supressora de Tumor p53/genética
13.
Eur Rev Med Pharmacol Sci ; 22(12): 3919-3924, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29949168

RESUMO

OBJECTIVE: The morbidity of atrial fibrillation (AF) is 1%-2% in clinic. Radiofrequency catheter ablation (RFCA) is a type of radical interventional therapy for AF, whereas it may lead to a hypercoagulable state. This study evaluated platelet particle CD62P and platelet activation biomarker GP IIb/IIIa expressions in AF patients treated by RFCA, and aimed to analyze their relationships with the hypercoagulable state after RFCA. PATIENTS AND METHODS: A total of 60 AF patients received RFCA in our hospital were enrolled. The patients were divided into group A as hypercoagulable state group and group B as non-hypercoagulable group. Healthy volunteers were selected as normal control. Serum D-Dimer, parathyroid activity index 1 (PAI-1), and tissue plasminogen activator (t-PA) content were tested by using enzyme-linked immunosorbent assay (ELISA), while peripheral CD62P and GP IIb/IIIa expressions were detected by using flow cytometry before, after, and seven days after RFCA. RESULTS: D-Dimer and PAI-1 levels increased, while t-PA reduced in group A compared with that in group B and control (p<0.05). D-Dimer and t-PA contents gradually elevated, whereas t-PA level gradually declined in group A before, after, and seven days after RFCA (p<0.05). Serum CD62P and GP IIb/IIIa expressions in group A were significantly higher compared to that in group B and control (p<0.05). CD62P and GP IIb/IIIa levels were significantly higher seven days after RFCA compared with immediate after RFCA in group A (p<0.05). CD62P showed a positive correlation with GP IIb/IIIa in hypercoagulable state patients after RFCA (p<0.05). CONCLUSIONS: AF patient may appear in hypercoagulable state after RFCA. CD62P and GP IIb/IIIa significantly increased and exhibited a positive correlation.


Assuntos
Fibrilação Atrial/cirurgia , Ablação por Cateter/efeitos adversos , Micropartículas Derivadas de Células/fisiologia , Selectina-P/fisiologia , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/fisiologia , Trombofilia/etiologia , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
14.
Sports Med ; 48(9): 2025-2039, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29868992

RESUMO

Initially suggested as simple cell debris, cell-derived microvesicles (MVs) have now gained acceptance as recognized players in cellular communication and physiology. Shed by most, and perhaps all, human cells, these tiny lipid-membrane vesicles carry bioactive agents, such as proteins, lipids and microRNA from their cell source, and are produced under orchestrated events in response to a myriad of stimuli. Physical exercise introduces systemic physiological challenges capable of acutely disrupting cell homeostasis and stimulating the release of MVs into the circulation. The novel and promising field of exercise-derived MVs is expanding quickly, and the following work provides a review of the influence of exercise on circulating MVs, considering both acute and chronic aspects of exercise and training. Potential effects of the MV response to exercise are highlighted and future directions suggested as exercise and sports sciences extend the realm of extracellular vesicles.


Assuntos
Exercício/fisiologia , Micropartículas Derivadas de Células/fisiologia , Humanos , MicroRNAs
15.
Am J Clin Nutr ; 107(6): 876-882, 2018 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-29741564

RESUMO

Background: Endothelial progenitor cells (EPCs) and microparticles are emerging as novel markers of cardiovascular disease (CVD) risk, which could potentially be modified by dietary fat. We have previously shown that replacing dietary saturated fatty acids (SFAs) with monounsaturated or n-6 (ω-6) polyunsaturated fatty acids (MUFAs or PUFAs, respectively) improved lipid biomarkers, blood pressure, and markers of endothelial activation, but their effects on circulating EPCs and microparticles are unclear. Objective: The Dietary Intervention and VAScular function (DIVAS) Study investigated the replacement of 9.5-9.6% of total energy (%TE) contributed by SFAs with MUFAs or n-6 PUFAs for 16 wk on EPC and microparticle numbers in United Kingdom adults with moderate CVD risk. Design: In this randomized, controlled, single-blind, parallel-group dietary intervention, men and women aged 21-60 y (n = 190) with moderate CVD risk (≥50% above the population mean) consumed 1 of three 16-wk isoenergetic diets. Target compositions for total fat, SFAs, MUFAs, and n-6 PUFAs (%TE) were as follows: SFA-rich diet (36:17:11:4; n = 64), MUFA-rich diet (36:9:19:4; n = 62), and n-6 PUFA-rich diet (36:9:13:10; n = 66). Circulating EPC, endothelial microparticle (EMP), and platelet microparticle (PMP) numbers were analyzed by flow cytometry. Dietary intake, vascular function, and other cardiometabolic risk factors were determined at baseline. Results: Relative to the SFA-rich diet, MUFA- and n-6 PUFA-rich diets decreased EMP (-47.3%, -44.9%) respectively and PMP (-36.8%, -39.1%) numbers (overall diet effects, P < 0.01). The MUFA-rich diet increased EPC numbers (+28.4%; P = 0.023). Additional analyses that used stepwise regression models identified the augmentation index (measuring arterial stiffness determined by pulse-wave analysis) as an independent predictor of baseline EPC and microparticle numbers. Conclusions: Replacement of 9.5-9.6%TE dietary SFAs with MUFAs increased EPC numbers, and replacement with either MUFAs or n-6 PUFAs decreased microparticle numbers, suggesting beneficial effects on endothelial repair and maintenance. Further studies are warranted to determine the mechanisms underlying the favorable effects on EPC and microparticle numbers after SFA replacement. This trial was registered at www.clinicaltrials.gov as NCT01478958.


Assuntos
Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/prevenção & controle , Micropartículas Derivadas de Células/fisiologia , Gorduras na Dieta/administração & dosagem , Células Progenitoras Endoteliais/fisiologia , Adulto , Biomarcadores , Estudos de Coortes , Gorduras na Dieta/classificação , Ácidos Graxos/administração & dosagem , Ácidos Graxos Insaturados/administração & dosagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Método Simples-Cego , Adulto Jovem
16.
PLoS One ; 13(4): e0195724, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29672621

RESUMO

BACKGROUND: Circulating endothelial microparticles (EMPs) and progenitor cells (PCs) are biological markers of endothelial function and endogenous repair capacity. The study was aimed to investigate whether COPD patients have an imbalance between EMPs to PCs compared to controls and to evaluate the effect of cigarette smoke on these circulating markers. METHODS: Circulating EMPs and PCs were determined by flow cytometry in 27 nonsmokers, 20 smokers and 61 COPD patients with moderate to severe airflow obstruction. We compared total EMPs (CD31+CD42b-), apoptotic if they co-expressed Annexin-V+ or activated if they co-expressed CD62E+, circulating PCs (CD34+CD133+CD45+) and the EMPs/PCs ratio between groups. RESULTS: COPD patients presented increased levels of total and apoptotic circulating EMPs, and an increased EMPs/PCs ratio, compared with nonsmokers. Women had less circulating PCs than men through all groups and those with COPD showed lower levels of PCs than both control groups. In smokers, circulating EMPs and PCs did not differ from nonsmokers, being the EMPs/PCs ratio in an intermediate position between COPD and nonsmokers. CONCLUSIONS: We conclude that COPD patients present an imbalance between endothelial damage and repair capacity that might explain the frequent concurrence of cardiovascular disorders. Factors related to the disease itself and gender, rather than cigarette smoking, may account for this imbalance.


Assuntos
Micropartículas Derivadas de Células/patologia , Endotélio Vascular/patologia , Doença Pulmonar Obstrutiva Crônica/sangue , Doença Pulmonar Obstrutiva Crônica/patologia , Idoso , Apoptose , Estudos de Casos e Controles , Micropartículas Derivadas de Células/fisiologia , Células Progenitoras Endoteliais/patologia , Células Progenitoras Endoteliais/fisiologia , Endotélio Vascular/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Regeneração/fisiologia , Testes de Função Respiratória , Fumar/sangue , Fumar/patologia , Fumar/fisiopatologia
17.
J Proteome Res ; 17(4): 1690-1699, 2018 04 06.
Artigo em Inglês | MEDLINE | ID: mdl-29494150

RESUMO

Circulating microvesicles are able to mediate long-distance cell-cell communications. It is essential to understand how microvesicles from pancreatic cancer act on other cells in the body. In this work, serum-derived microvesicles were isolated from 10 patients with locally advanced pancreatic cancer and healthy controls. Using Cell Transwell and WST-1 reagents, we found that microvesicles from pancreatic cancer accelerated migration and proliferation of PANC-1 cells. Meanwhile, the proliferation of these cancer-microvesicle-treated cells (CMTCs) was affected less by 10 µM of gemcitabine relative to healthy microvesicle-treated cells (HMTCs). Next, we optimized the filter-aided sample preparation method to increase the recovery of protein samples and then applied it to the quantification of the proteome of CMTCs and HMTCs. The peptides were labeled and analyzed by liquid chromatography-tandem mass spectrometry. In total, 4102 proteins were identified, where 35 proteins were up-regulated with 27 down-regulated in CMTCs. We verified the quantitative results of three key proteins CD44, PPP2R1A, and TP53 by Western blot. The Ingenuity Pathway Analysis revealed pathways that cancer microvesicles might participate in to promote cell migration and proliferation. These findings may provide novel clues of treatment for tumorigenesis and metastasis.


Assuntos
Micropartículas Derivadas de Células/transplante , Neoplasias Pancreáticas/patologia , Antimetabólitos Antineoplásicos/farmacologia , Estudos de Casos e Controles , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Micropartículas Derivadas de Células/fisiologia , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Regulação da Expressão Gênica , Humanos , Receptores de Hialuronatos/análise , Proteína Fosfatase 2/análise , Proteoma/análise , Proteína Supressora de Tumor p53/análise
18.
PLoS One ; 13(3): e0194004, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29538408

RESUMO

Extracellular vesicles (EVs) released by virtually every cell of all organisms are involved in processes of intercellular communication through the delivery of their functional mRNAs, proteins and bioactive lipids. We previously demonstrated that mouse embryonic stem cell-released EVs (mESEVs) are able to transfer their content to different target retinal cells, inducing morphological and biochemical changes in them. The main objective of this paper is to characterize EVs derived from human embryonic stem cells (hESEVs) and investigate the effects that they have on cultured retinal glial, progenitor Müller cells, which are known to give rise to retinal neurons under specific conditions. This would allow us to establish if hESEVs have a pro-regenerative potential not yet described that could be used in the future for treatment of human retinal degenerative diseases. Initially, we showed that hESEVs are heterogeneous in size, contain mRNAs and proteins involved in the induction and maintenance of stem cell pluripotency and can be internalized by cultured Müller cells. After a single exposure to hESEVs these cells display changes in their gene expression profile, and with multiple exposures they de-differentiate and trans-differentiate into retinal neuronal precursors. hESEVs were then fractionated into microvesicles (MVs) and exosomes (EXOs), which were characterized by size, specific surface proteins and biochemical/molecular components. We demonstrate that despite the similar internalization of non-fractionated hESEVs, MVs and EXOs by Müller progenitor cells, in vitro, only the release of MVs' cargo into the cells' cytoplasm induces specific changes in their levels of pluripotency mRNAs and early retinal proteins. EXOs do not produce any detectable effect. Thus, we conclude that MVs and MVs-containing hESEVs are promising agents that possibly could promote the regeneration of diseased or damaged retinas in vivo through inducing glial Müller cells to become replacement neurons.


Assuntos
Células Ependimogliais/fisiologia , Vesículas Extracelulares/fisiologia , Células-Tronco Embrionárias Humanas/fisiologia , Micropartículas Derivadas de Células/metabolismo , Micropartículas Derivadas de Células/fisiologia , Células Cultivadas , Células Ependimogliais/metabolismo , Exossomos/metabolismo , Exossomos/fisiologia , Vesículas Extracelulares/metabolismo , Células HEK293 , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Neuroglia/metabolismo , Neuroglia/fisiologia , Neurônios/metabolismo , Neurônios/fisiologia , Células-Tronco Pluripotentes/metabolismo , Células-Tronco Pluripotentes/fisiologia , RNA Mensageiro/metabolismo , Regeneração/fisiologia , Retina/metabolismo , Retina/fisiologia , Transcriptoma/fisiologia
19.
Curr Biol ; 28(4): R170-R185, 2018 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-29462587

RESUMO

The maintenance of a healthy and functional mitochondrial network is critical during development as well as throughout life in the response to physiological adaptations and stress conditions. Owing to their role in energy production, mitochondria are exposed to high levels of reactive oxygen species, making them particularly vulnerable to mitochondrial DNA mutations and protein misfolding. Given that mitochondria are formed from proteins encoded by both nuclear and mitochondrial genomes, an additional layer of complexity is inherent in the coordination of protein synthesis and the mitochondrial import of nuclear-encoded proteins. For these reasons, mitochondria have evolved multiple systems of quality control to ensure that the requisite number of functional mitochondria are present to meet the demands of the cell. These pathways work to eliminate damaged mitochondrial proteins or parts of the mitochondrial network by mitophagy and renew components by adding protein and lipids through biogenesis, collectively resulting in mitochondrial turnover. Mitochondrial quality control mechanisms are multi-tiered, operating at the protein, organelle and cell levels. Herein, we discuss mitophagy in different physiological contexts and then relate it to other quality control pathways, including the unfolded protein response, shedding of vesicles, proteolysis, and degradation by the ubiquitin-proteasome system. Understanding how these pathways contribute to the maintenance of mitochondrial homeostasis could provide insights into the development of targeted treatments when these systems fail in disease.


Assuntos
Homeostase/fisiologia , Mitocôndrias/fisiologia , /fisiologia , Animais , Micropartículas Derivadas de Células/fisiologia , Humanos , Complexo de Endopeptidases do Proteassoma/fisiologia , Proteólise , Ubiquitina/fisiologia , Resposta a Proteínas não Dobradas/fisiologia
20.
Circ Res ; 122(3): 506-522, 2018 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-29420211

RESUMO

Platelets play a vital role in normal hemostasis to stem blood loss at sites of vascular injury by tethering and adhering to sites of injury, recruiting other platelets and blood cells to the developing clot, releasing vasoactive small molecules and proteins, and assembling and activating plasma coagulation proteins in a tightly regulated temporal and spatial manner. In synchrony with specific end products of coagulation, primarily cross-linked fibrin, a stable thrombus quickly forms. Far beyond physiological hemostasis and pathological thrombosis, emerging evidence supports platelets playing a pivotal role in vascular homeostasis, inflammation, cellular repair, regeneration, and wide range of autocrine and paracrine functions. In essence, platelets play both structural and functional roles as reporters, messengers, and active transporters surveying the vasculature for cues of environmental or developmental stimuli and participating as first responders.1 In this review, we will provide a contemporary perspective of platelet physiology, including fundamental, translational, and clinical constructs that apply directly to human health and disease.


Assuntos
Plaquetas/fisiologia , Hemostasia/fisiologia , Animais , Aterosclerose/sangue , Aterosclerose/fisiopatologia , Proteínas Sanguíneas/fisiologia , Micropartículas Derivadas de Células/fisiologia , Sistemas de Liberação de Medicamentos , Desenvolvimento de Medicamentos , Endotélio Vascular/fisiologia , Proteínas da Matriz Extracelular/fisiologia , Armadilhas Extracelulares/fisiologia , Interações Hospedeiro-Patógeno , Humanos , /fisiopatologia , Inflamação/sangue , Inflamação/fisiopatologia , Linfangiogênese , MicroRNAs/sangue , Neovascularização Fisiológica , Inibidores da Agregação de Plaquetas/uso terapêutico , Glicoproteínas da Membrana de Plaquetas/fisiologia , Plasma Rico em Plaquetas , Regeneração/fisiologia , Tromboembolia/sangue , Tromboembolia/tratamento farmacológico , Tromboembolia/fisiopatologia , Trombopoese
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