Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.017
Filtrar
1.
Ecotoxicol Environ Saf ; 208: 111730, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396061

RESUMO

Copper (Cu) is a common environmental pollutant in nature. Cu-poisoning can cause liver damage and erythrocytes hemolysis. To evaluate the effect of CuSO4 poisoning on the morphological and functional characteristics of goat red blood cells. Five 10-14-month-old goats were selected for jugular vein blood sampling to obtain erythrocytes, and then the erythrocytes were processed with different concentrations (0, 10, 20, 30, 40 and 50 µmol/L) of CuSO4 for 48 h, and 40 µmol/L doses CuSO4 incubated for different time (12, 24, 36, 48 and 60 h) to process erythrocytes. We observed the changes in erythrocyte morphology through scanning electron microscopy, and detected the antioxidant function and activities of three ATPases. Additionally, biological properties were examined from the perspectives of phospholipids and membrane protein components, permeability fragility, and fluidity in erythrocytes. We found that after CuSO4 treatment, the antioxidant capacity of erythrocytes decreased, which was manifested as increased MDA content and decreased CuZn-SOD and GSH-Px activities (p < 0.05). In addition, we also found that erythrocyte fluidity decreased, osmotic fragility increased, membrane phospholipid percentage and protein composition changes abnormally, and Na+/K+-ATPase, Mg2+-ATPase and Ca2+-ATPase activities decreased (p < 0.05). From the results, it can be concluded that CuSO4 exposure causes hemolysis of goat erythrocytes through oxidative stress to the structure and function of erythrocytes, showing a dose-time effect.


Assuntos
Sulfato de Cobre/toxicidade , Substâncias Perigosas/toxicidade , Adenosina Trifosfatases/metabolismo , Animais , Antioxidantes/metabolismo , Cobre/análise , Membrana Eritrocítica/química , Membrana Eritrocítica/efeitos dos fármacos , Membrana Eritrocítica/metabolismo , Eritrócitos/efeitos dos fármacos , Cabras/metabolismo , Hemólise/efeitos dos fármacos , Fragilidade Osmótica/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fosfolipídeos/análise , Testes de Toxicidade
2.
Int J Nanomedicine ; 15: 6749-6760, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32982231

RESUMO

Background: The combination of radiotherapy (RT) and chemotherapy, as a standard treatment for breast cancer in the clinic, is unsatisfactory due to chemoradioresistance and severe side effects. Methods and Results: To address these issues, a cancer cell-erythrocyte hybrid membrane-coated doxorubicin (DOX)-loaded gold nanocage (CM-EM-GNCs@DOX) was constructed for near-infrared light (NIR)-activated photothermal/radio/chemotherapy of breast cancer. CM-EM-GNCs@DOX inherited an excellent homologous target ability from the cancer cell membrane and an immune evasion capability from the erythrocyte membrane, together resulting in highly efficient accumulation in the tumor site with decreased clearance. Following the highly efficient uptake of CM-EM-GNCs@DOX in cancer cells, the RT efficacy was remarkably amplified due to the radiosensitization effect of CM-EM-GNCs@DOX, which reduced the needed radiotherapeutic dose. Importantly, with NIR irradiation, CM-EM-GNCs@DOX exerted a high photothermal effect, which not only ruptured CM-EM-GNCs@DOX to release DOX for precise and controllable chemotherapy, but also potentiated chemo/radiotherapy by photothermal therapy. Conclusion: Therefore, a highly efficient and safe combined photothermal/radio/chemotherapy approach was achieved in vitro and in vivo by CM-EM-GNCs@DOX, which provided a promising strategy for treating breast cancer.


Assuntos
Neoplasias da Mama/terapia , Membrana Celular/química , Doxorrubicina/administração & dosagem , Nanoestruturas/química , Fototerapia/métodos , Animais , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/farmacocinética , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Doxorrubicina/farmacocinética , Membrana Eritrocítica/química , Feminino , Ouro/química , Humanos , Hipertermia Induzida/métodos , Raios Infravermelhos , Células MCF-7 , Fusão de Membrana , Camundongos , Camundongos Nus , Nanoestruturas/administração & dosagem , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/farmacologia , Células RAW 264.7 , Ensaios Antitumorais Modelo de Xenoenxerto
3.
Biochim Biophys Acta Biomembr ; 1862(7): 183309, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32298678

RESUMO

The decrease in cellular deformability shows strong correlation with erythrocyte aging. Cell deformation can be divided into passive deformation and active deformation; however, the active deformation has been ignored in previous studies. In this work, Young's moduli of age-related erythrocytes were tested by atomic force microscopy. Furthermore, the deformation and passive and active deformation values were calculated by respective areas. Our results showed that erythrocytes in the densest fraction had the highest values of the Young's modulus, deformation, and active deformation, but the lowest values of passive deformation. Moreover, values of the deformation and active deformation both increased gradually with erythrocyte aging. The present data indicate that the elastic hysteresis loop between the approach and the retract curve could be regarded as erythrocyte deformability, and cellular deformability could be characterized by energy states. In addition, active deformation might be a crucial mechanical factor for clearing aged erythrocytes. This could provide an important information on erythrocyte biomechanics in the removal of aged cell.


Assuntos
Envelhecimento Eritrocítico/fisiologia , Deformação Eritrocítica/fisiologia , Membrana Eritrocítica/ultraestrutura , Eritrócitos/ultraestrutura , Membrana Eritrocítica/química , Eritrócitos/fisiologia , Humanos , Microscopia de Força Atômica
4.
Biomater Sci ; 8(7): 1802-1814, 2020 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-32163070

RESUMO

Upconversion nanoparticles (UCNPs) have been widely employed for tumor imaging using magnetic resonance imaging (MRI) and upconversion luminescence (UCL) imaging. The short blood clearance time and immunogenicity of UCNPs have limited their further application in vivo. We have designed UCNPs camouflaged with an exterior red blood cell (RBC) membrane coating (RBC-UCNPs) to solve these problems. Moreover, because of some intrinsic disadvantages of MRI and UCL imaging, we investigated the use of pretargeted RBC-UCNPs for positron-emission tomography (PET) imaging to obtain more comprehensive information. Our data showed that RBC-UCNPs retained the immunity feature from the source cells and the superior optical and chemical features from the pristine UCNP cores. The tumor-targeting ability of RBC-UCNPs was enhanced by binding 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[folate(polyethylene glycol)-2000] (DSPE-PEG-FA) molecules onto the cell membranes. PET imaging with short half-life radionuclides to visualize the RBC-UCNPs was successfully realized by a combination of pre-targeting and in vivo click chemistry. Blood chemistry, hematology, and histologic analysis suggested good in vivo biocompatibility of the RBC-UCNPs. Our method provides a new potential biomedical application of biomimetic nanoparticles.


Assuntos
Membrana Eritrocítica/química , Ácido Fólico/química , Fosfatidiletanolaminas/química , Polietilenoglicóis/química , Neoplasias de Mama Triplo Negativas/diagnóstico por imagem , Animais , Materiais Biomiméticos/química , Linhagem Celular Tumoral , Química Click , Feminino , Fluoretos/química , Gadolínio/química , Humanos , Imagem por Ressonância Magnética , Camundongos , Imagem Multimodal , Nanopartículas/química , Transplante de Neoplasias , Tomografia por Emissão de Pósitrons , Itérbio/química
5.
Dalton Trans ; 49(8): 2645-2651, 2020 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-32040099

RESUMO

Photothermal therapy (PTT), which involves an increase in temperature triggered only by light signals at tumor sites to remove cancer cells, has been considered an attractive strategy in cancer therapy. Nevertheless, the in vivo applications of photosensitizer-based PTT are limited due to the poor biocompatibility of photothermal agents. Employing red blood cell (RBC) membranes to encapsulate photothermal agents can solve this issue, but the extra surface coating will suppress heat dissipation, which is unfavorable for the subsequent treatment. Herein, biomimetic nano-capsules have been fabricated for light signal-activated cancer therapy by encapsulating photocatalyst titanium dioxide colloid and photothermal agent gold nanorods (Au NRs) in erythrocyte membrane vesicles. The fabricated Au/TiO2@RBC nano-capsules can achieve the controlled release of Au NRs upon the photocatalytic degradation of their surface cell membrane coatings, and generate therapeutic signals after the released Au NRs are irradiated by an NIR laser. Meanwhile, the reactive oxygen species (ROS) produced by photocatalysis are helpful for killing tumor cells photodynamically. Thus, the biomimetic nano-capsules prepared herein will contribute to the research and development in cancer cell therapy.


Assuntos
Neoplasias da Mama/terapia , Membrana Eritrocítica/química , Hipertermia Induzida , Nanopartículas Metálicas/administração & dosagem , Fotoquimioterapia , Fármacos Fotossensibilizantes/administração & dosagem , Fototerapia , Biomimética , Cápsulas , Proliferação de Células , Feminino , Ouro/química , Humanos , Nanopartículas Metálicas/química , Espécies Reativas de Oxigênio , Titânio/química , Células Tumorais Cultivadas
6.
Nanoscale ; 12(6): 4137-4149, 2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-32022084

RESUMO

Recent progress in bioimaging nanotechnology has a great impact on the diagnosis, treatment, and prevention of diseases by enabling early intervention. Among different types of bioimaging modalities, contrast-enhanced magnetic resonance imaging using paramagnetic gadolinium-based molecular contrast agents (GBCAs) are most commonly used in clinic. However, molecular GBCAs distribute rapidly between plasma and interstitial spaces with short half-lives limiting its clinical impacts. To improve the properties of GBCAs, herein an effort has been put forth by incorporating GBCA into nanoscale system mimicking the property of red blood cell (RBC) that could facilitate contrast enhancement and prolong intraluminal retention in the body. The proposed nanoconstruct is made up of polymeric-core labeled with lipid conjugated GBCA followed by the imprint of the RBC membrane concealment layer to enhance stability and biocompatibility. Meanwhile, the confinement strategy of GBCA was implemented to accelerate magnetic properties of nanoconstruct providing longitudinal-relaxivity (r1) to 12.78 ± 0.29 (mM s)-1. Such improvement in r1 was further confirmed by enhanced contrast in the vascular angiography of the murine model. Given higher colloidal stability and tunable magnetic properties, nanoconstruct proposed herein is a promising platform technology for the applications where enhanced plasma residence time and magnetic properties are necessary for diagnosis and therapy.


Assuntos
Meios de Contraste/química , Membrana Eritrocítica/química , Imagem por Ressonância Magnética/métodos , Nanopartículas/química , Animais , Meios de Contraste/farmacocinética , Feminino , Gadolínio/química , Gadolínio/farmacocinética , Humanos , Camundongos , Camundongos Nus , Células THP-1
7.
Biochim Biophys Acta Biomembr ; 1862(5): 183188, 2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-31930963

RESUMO

Langmuir films prepared from bovine erythrocyte membranes (LFBEM) were studied and transferred to alkylated glasses (Langmuir-Blodgett films, LBBEM) in order to assess the effects of membrane molecular packing on Bovine Erythrocyte Acetylcholinesterase (BEA) catalytic activity. Surface pressure (π) vs Area isotherms showed three 2D-transitions at ~7, ~18 and ~44 mN/m and a collapse pressure at πc = 49 mN/m. The 0-12-0 mN/m compression-decompression cycles resulted reversible while those 0-40-0 mN/m exhibited a significant hysteresis. Taken together, EFM, BAM and AFM images and the stability of the film after 3C-D cycles, we can suggest that over the air-water interface as well as over the silanized glass substrate the surface is mostly covered by a monolayer with a few particles dispersed. Acetylthiocholine hydrolysis was assayed with BEA in bovine erythrocyte membrane suspensions (SBEM) and in LBBEM packed at 10 (LBBEM,10) and 35 mN/m (LBBEM,35), which gave the following kinetic parameters: Vmax = 3.41 ± 0.15, 0.021 ± 0.002 and 0.030 ± 0.003 nmol.min-1·µg prot-1 and KM = 0.11 ± 0.02, 0.047 ± 0.017 and 0.026 ± 0.017 mM, respectively. Although from SBEM to LBBEM we lost active enzyme, the catalytic efficiency (Vmax/KM) increased ~750 times. Eugenol and 1,8-cineol inhibited BEA catalytic activity in LBBEM,35. Our results demonstrate the transmission of information between the membrane and the environment within the subphase immediately below the membrane, where anchored proteins are hosted. This was reflected by the membrane packing-induced modulation of BEA catalytic activity. Furthermore, LBBEM provides a proof of concept for the development of biosensors to screen new green pesticides acting through BEA interaction.


Assuntos
Acetilcolinesterase/metabolismo , Membrana Eritrocítica/química , Membrana Eritrocítica/metabolismo , Acetilcolinesterase/química , Acetilcolinesterase/fisiologia , Adsorção/fisiologia , Animais , Catálise , Bovinos , Membrana Eritrocítica/fisiologia , Hidrólise , Cinética , Microscopia de Força Atômica/métodos , Estudo de Prova de Conceito , Propriedades de Superfície , Água/química
8.
Artigo em Inglês | MEDLINE | ID: mdl-31926297

RESUMO

Maternal smoking-induced congenital heart and microvascular defects are closely associated with the impaired functioning of the in-utero feto-placental circulation system. Current groundbreaking facts revealed intimate crosstalk between circulating red blood cells (RBCs) and the vascular endothelium. Thus, RBCs have become the protagonists under varied pathological and adverse pro-oxidative cellular stress conditions. We isolated and screened fetal RBCs from the arterial cord blood of neonates, born to non-smoking (RBC-NS) and smoking mothers (RBC-S), assuming that parameters of fetal RBCs are blueprints of conditions experienced in-utero. Using atomic force microscopy and mass spectrometry-based shotgun lipidomics in the RBC-S population we revealed induced membrane stiffness, loss in intrinsic plastic activities and several abnormalities in their membrane-lipid composition, that could consequently result in perturbed hemodynamic flow movements. Altogether, these features are indicative of the outcome of neonatal microvascular complications and suggest unavailability for the potential rescue mechanism in cases of vascular endothelium impairment due to altered membrane integrity and rheological properties.


Assuntos
Eritrócitos/patologia , Sangue Fetal/citologia , Efeitos Tardios da Exposição Pré-Natal/etiologia , Poluição por Fumaça de Tabaco/efeitos adversos , Adulto , Fenômenos Biomecânicos , Membrana Eritrocítica/química , Membrana Eritrocítica/patologia , Eritrócitos/química , Feminino , Hemodinâmica , Humanos , Recém-Nascido , Peroxidação de Lipídeos , Fluidez de Membrana , Lipídeos de Membrana/análise , Gravidez , Efeitos Tardios da Exposição Pré-Natal/patologia , Adulto Jovem
9.
Biochim Biophys Acta Biomembr ; 1862(2): 183067, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31634445

RESUMO

In this paper, systematic studies concerning the influence of selected oxysterols on the structure and fluidity of human erythrocyte membrane modeled as Langmuir monolayers have been performed. Three oxidized cholesterol derivatives, namely 7α-hydroxycholesterol (7α-OH) 7ß-hydroxycholesterol (7ß-OH) and 7-ketocholesterol (7-K) have been incorporated in two different proportions (10 and 50%) into artificial erythrocyte membrane, modeled as two-component (cholesterol:POPC) Langmuir monolayer. All the studied oxysterols were found to alter membrane fluidity and the effect was more pronounced for higher oxysterol content. 7α-OH increased membrane fluidity while opposite effect was observed for 7ß-OH and 7-K. Experiments performed on model systems have been verified in biological studies on red blood cells (RBC). Consistent results have been found, i.e. under the influence of 7α-OH, the elasticity of erythrocytes increased, and in the presence of other investigated oxysterols - decreased. The strongest effect was noticed for 7-K. Change of membrane elasticity was associated with the change of erythrocytes shape, being most noticeable under the influence of 7-K.


Assuntos
Membrana Eritrocítica/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Oxisteróis/farmacologia , Forma Celular/efeitos dos fármacos , Células Cultivadas , Elasticidade/efeitos dos fármacos , Membrana Eritrocítica/química , Humanos , Hidroxicolesteróis/farmacologia , Cetocolesteróis/farmacologia , Fluidez de Membrana/efeitos dos fármacos , Membranas Artificiais , Oxisteróis/química , Fosfatidilcolinas
10.
Biochim Biophys Acta Biomembr ; 1862(2): 183126, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31738902

RESUMO

Over the past few decades, studies on the red blood cell (RBC) membrane gave rise to increasingly sophisticated although divergent models of its structural organization, since investigations were often performed in denaturing conditions using detergents. To access soluble isolated RBC membrane complexes with the preservation of their interactions and conformations, we decided to apply the recent SMALP (Styrene Maleic Acid Lipid Particles) technology to RBC ghosts. Depending on the ionic strength of buffers in which ghost membranes were resuspended, the isolated proteins within SMALPs could differ on Coomassie-stained gels, but with few changes when compared to ghost membrane SDS lysates. We subsequently produced SMALPs derived from ghosts from two different blood group phenotypes, RhD-positive and RhD-negative, both types of RBC expressing the RhCE proteins but only RhD-positive cells being able to express the RhD proteins. This allowed the isolation, by size exclusion chromatography (SEC), of soluble fractions containing the Rh complex, including the RhD protein or not, within SMALPs. The use a conformation-dependent anti-RhD antibody in immunoprecipitation studies performed on SEC fractions of SMALPs containing Rh proteins clearly demonstrated that the RhD protein, which was only present in SMALPs prepared from RhD-positive RBC ghosts, has preserved at least one important conformational RhD epitope. This approach opens new perspectives in the field of the erythroid membrane study, such as visualization of RBC membrane complexes in native conditions by cryo-electron microscopy (CryoEM) or immuno-tests with conformation-dependent antibodies against blood group antigens on separated and characterized SMALPs containing RBC membrane proteins.


Assuntos
Membrana Eritrocítica/química , Detergentes/química , Membrana Eritrocítica/imunologia , Humanos , Lipossomos/química , Maleatos/química , Proteínas Recombinantes de Fusão/imunologia , Estirenos/química
11.
J Med Food ; 23(1): 37-42, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31855493

RESUMO

The association between obesity and erythrocyte fatty acids (FAs) has been suggested; however, there have been no studies on the effects of onion peel extract (OPE) on the composition of erythrocyte FAs. This study aimed to investigate the effects of OPE on the composition of erythrocyte FAs in overweight and obese subjects. This was a randomized, double-blind, and placebo-controlled trial conducted in overweight and obese Korean subjects. The placebo and OPE groups were taking placebo capsule or OPE capsule twice per day for 12 weeks. Body composition and fat distribution were measured using dual-energy X-ray absorptiometry. The OPE group showed significantly reduced body weight, body mass index, body fat mass, and percentage of body fat mass. After 12 weeks, eicosapentaenoic acid and monounsaturated FAs of the placebo group were significantly lower at baseline. Consumption of OPE ameliorated the decreasing polyunsaturated n-3 polyunsaturated FA (PUFA) n-3 and increasing PUFA n-6, which prevented an increased n-6/n-3 ratio. The changes in arm fat percentage (ARFATP), trunk fat percentage, and total fat percentage (FATP) were negatively correlated with the change in PUFA n-3. In addition, increased erythrocyte docosahexaenoic acid was associated with decreased ARFATP and FATP. These results suggest that OPE has beneficial effects on obesity by regulating erythrocyte n-6/n-3 ratio and preventing fat accumulation in various body regions.


Assuntos
Membrana Eritrocítica/química , Ácidos Graxos Ômega-3/química , Obesidade/sangue , Cebolas/química , Sobrepeso/sangue , Extratos Vegetais/farmacologia , Adiposidade , Adulto , Índice de Massa Corporal , Peso Corporal , Método Duplo-Cego , Ácidos Graxos Ômega-6/química , Humanos , Pessoa de Meia-Idade , República da Coreia
12.
Georgian Med News ; (294): 103-108, 2019 Sep.
Artigo em Russo | MEDLINE | ID: mdl-31687959

RESUMO

The article overviews some issues of the severe course of tropical malaria. In addition to the analysis of the ongoing situation with malaria in Russia, a general clinical picture of the severe course of tropical malaria is discussed. The main part of the overview includes a detailed analysis of current data on the molecular genetic aspects of the erythrocytes' adhesion in the case of tropical malaria. The main elements involved in the process of binding red blood cells and, as a result, in the process of their adhesion to other cells of the human body were considered in detail. Data were studied and summarized not only on protein interactions between an infected red cell and its cellular environment, but also on the genetic characteristics of the parasite leading to similar molecular-biological processes. In addition to the study of protein PfEMP1 role which is nowadays well-considered in the literature, the most up-to-date but less reported data on erythrocyte adhesion proteins STEVOR and RIFIN were also included. The team of authors hopes that this publication will help to get a deeper insight into the problem of erythrocyte adhesion in the course of complicated malaria infection forms and to summarize some of the available data on this issue.


Assuntos
Antígenos de Protozoários/metabolismo , Membrana Eritrocítica/química , Membrana Eritrocítica/parasitologia , Eritrócitos/parasitologia , Malária Falciparum , Proteínas de Membrana/metabolismo , Plasmodium falciparum/metabolismo , Proteínas de Protozoários/metabolismo , Antígenos de Protozoários/química , Humanos , Malária Falciparum/sangue , Proteínas de Membrana/química , Plasmodium falciparum/fisiologia , Proteínas de Protozoários/química , Federação Russa
13.
J Control Release ; 314: 81-91, 2019 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-31644936

RESUMO

Erythrocyte-mimicking nanovehicles (EM-NVs) are developed by fusing nanoparticle cores with naturally derived erythrocyte membranes. Compared with conventional nanosystems, EM-NVs hold preferable characteristics of prolonged blood circulation time and immune evasion. Due to the cell surface mimetic properties, along with tailored core material, EM-NVs have huge application potential in a large variety of biomedical fields, which are anticipated to revolutionize the present theranostic modalities of diseases in clinic. This review focuses on (I) drug carriers, (II) photosensitizers, (III) antidotes, (IV) vaccines and (V) probes, aiming to present an overall summary of the latest advancement in the application of EM-NVs, and highlight the major challenges and opportunities in this field.


Assuntos
Materiais Biomiméticos/química , Eritrócitos/química , Nanopartículas , Animais , Portadores de Fármacos/química , Membrana Eritrocítica/química , Humanos , Propriedades de Superfície , Nanomedicina Teranóstica/métodos
14.
Blood Adv ; 3(17): 2653-2663, 2019 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-31506283

RESUMO

The capacity to undergo substantial deformation is a defining characteristic of the red blood cell (RBC), facilitating transit through the splenic interendothelial slits and microvasculature. Establishment of this remarkable property occurs during a process of reticulocyte maturation that begins with egress through micron-wide pores in the bone marrow and is completed within the circulation. The requirement to undertake repeated cycles of deformation necessitates that both reticulocytes and erythrocytes regulate membrane-cytoskeletal protein interactions in order to maintain cellular stability. In the absence of transcriptional activity, modulation of these interactions in RBCs is likely to be achieved primarily through specific protein posttranslational modifications, which at present remain undefined. In this study, we use high-throughput methods to define the processes that underlie the response to deformation and shear stress in both reticulocytes and erythrocytes. Through combination of a bead-based microsphiltration assay with phosphoproteomics we describe posttranslational modification of RBC proteins associated with deformation. Using microsphiltration and microfluidic biochip-based assays, we explore the effect of inhibiting kinases identified using this dataset. We demonstrate roles for GSK3 and Lyn in capillary transit and maintenance of membrane stability following deformation and show that combined inhibition of these kinases significantly decreases reticulocyte capacity to undergo repeated deformation. Finally, we derive a comprehensive and integrative phosphoproteomic dataset that provides a valuable resource for further mechanistic dissection of the molecular pathways that underlie the RBC's response to mechanical stimuli and for the study of reticulocyte maturation.


Assuntos
Deformação Eritrocítica/fisiologia , Eritrócitos/fisiologia , Proteínas de Membrana/metabolismo , Fosforilação/fisiologia , Forma Celular , Células Cultivadas , Membrana Eritrocítica/química , Membrana Eritrocítica/metabolismo , Eritrócitos/citologia , Quinase 3 da Glicogênio Sintase/metabolismo , Humanos , Processamento de Proteína Pós-Traducional/fisiologia , Proteômica , Reticulócitos/citologia , Reticulócitos/fisiologia , Quinases da Família src/metabolismo
15.
Br J Haematol ; 187(1): 13-24, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31364155

RESUMO

Hereditary erythrocyte membrane disorders are caused by mutations in genes encoding various transmembrane or cytoskeletal proteins of red blood cells. The main consequences of these genetic alterations are decreased cell deformability and shortened erythrocyte survival. Red blood cell membrane defects encompass a heterogeneous group of haemolytic anaemias caused by either (i) altered membrane structural organisation (hereditary spherocytosis, hereditary elliptocytosis, hereditary pyropoikilocytosis and Southeast Asian ovalocytosis) or (ii) altered membrane transport function (overhydrated hereditary stomatocytosis, dehydrated hereditary stomatocytosis or xerocytosis, familial pseudohyperkalaemia and cryohydrocytosis). Herein we provide a comprehensive review of the recent literature on the molecular genetics of erythrocyte membrane defects and their reported clinical consequences. We also describe the effect of low-expression genetic variants on the high inter- and intra-familial phenotype variability of erythrocyte structural defects.


Assuntos
Anemia Hemolítica Congênita/genética , Membrana Eritrocítica/química , Alelos , Anemia Hemolítica Congênita/sangue , Anemia Hemolítica Congênita/diagnóstico , Eliptocitose Hereditária/sangue , Eliptocitose Hereditária/diagnóstico , Eliptocitose Hereditária/genética , Eritrócitos Anormais/metabolismo , Humanos , Proteínas de Membrana/sangue , Esferocitose Hereditária/sangue , Esferocitose Hereditária/diagnóstico , Esferocitose Hereditária/genética
16.
Cells ; 8(8)2019 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-31412631

RESUMO

Recently, biomimetic nanoparticles, especially cell membrane-cloaked nanoparticles, have attracted increasing attention in biomedical applications, including antitumor therapy, detoxification, and immune modulation, by imitating the structure and the function of biological systems such as long circulation life in the blood. However, the circulation time of cell membrane-cloaked nanoparticles is far less than that of the original cells, greatly limiting their biomedical applications, while the underlying reasons are seldom demonstrated. In this study, the influence of particle size on the circulation and the biodistribution of red blood cell membrane-coated nanoparticles (RBC-NPs) as model biomimetic nanoparticles were investigated. Differently sized RBC-NPs (80, 120, 160, and 200 nm) were prepared by fusing RBC membranes on poly(lactic-co-glycolic acid) nanoparticles. It was shown that the particle size did not change the cellular uptake of these biomimetic nanoparticles by macrophage cells in vitro and their immunogenic responses in vivo. However, their circulation life in vivo decreased with the particle size, while their accumulation in the liver increased with the particle size, which might be related to their size-dependent filtration through hepatic sinusoids. These findings will provide experimental evidence for the design and the optimization of biomimetic nanoparticles.


Assuntos
Materiais Biomiméticos/farmacocinética , Materiais Revestidos Biocompatíveis/farmacocinética , Nanopartículas/química , Tamanho da Partícula , Animais , Sistemas de Liberação de Medicamentos , Membrana Eritrocítica/química , Masculino , Camundongos , Camundongos Endogâmicos ICR , Células RAW 264.7 , Distribuição Tecidual
17.
Biochim Biophys Acta Proteins Proteom ; 1867(11): 140267, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31470132

RESUMO

Spectrin, the major protein component of the erythrocyte membrane skeleton has chaperone like activity and is known to bind membrane phospholipids and hemoglobin. We have probed the chaperone activity of spectrin in presence of hemoglobin and phospholipid SUVs of different compositions to elucidate the effect of phospholipid/hemoglobin binding on chaperone function. It is seen that spectrin displays a preference for hemoglobin over other substrates leading to a decrease in chaperone activity in presence of hemoglobin. A competition is seen to exist between phospholipid binding and chaperone function of spectrin, in a dose dependent manner with the greatest extent of decrease being seen in case of phospholipid vesicles containing aminophospholipids e.g. PS and PE which may have implications in diseases like hereditary spherocytosis where mutation in spectrin is implicated in its detachment from cell membrane. To gain a clearer understanding of the chaperone like activity of spectrin under in-vivo like conditions we have investigated the effect of macromolecular crowders as well as phosphorylation and glycation states on chaperone activity. It is seen that the presence of non-specific, protein and non-protein macromolecular crowders do not appreciably affect chaperone function. Phosphorylation also does not affect the chaperone function unlike glycation which progressively diminishes chaperone activity. We propose a model where chaperone clients adsorb onto spectrin's surface and processes that bind to and occlude these surfaces decrease chaperone activity.


Assuntos
Membrana Eritrocítica/química , Hemoglobinas/química , Chaperonas Moleculares/química , Espectrina/química , Animais , Bovinos , Membrana Eritrocítica/metabolismo , Hemoglobinas/metabolismo , Chaperonas Moleculares/metabolismo , Fosfolipídeos/química , Fosfolipídeos/metabolismo , Ovinos , Espectrina/metabolismo
18.
Anal Chim Acta ; 1080: 189-195, 2019 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-31409469

RESUMO

Direct and absolute analysis of microRNAs (miRNAs) in complex media (e.g., human serum) is still a big challenge due to the issues with off-analyte absorption, low sensitivity and specificity. In this work, we have fabricated the erythrocyte membrane-biointerfaced spherical nucleic acids (EMSNAs) for miRNA assay, which not only enables tailor-engineered signal amplification but also exhibits anti-interference property. As a consequence, it is possible to achieve a single-step quantification of miRNAs in complex media without the process of enzymatic amplification, which can vastly simplify the detection procedure. Experimental results reveal that the assay permits ultrasensitive quantification of miR-141, with a limit of detection down to 33.9 aM, and show a high selectivity for discriminating miR-200 family members. More importantly, the assay enables robust miRNA analysis in human serum and can accurately differentiate lung cancer patients and prostate cancer patients from healthy donors. Its performance may satisfy the requirements for direct, rapid, sensitive and specific early diagnosis of cancer, signifying its great potential in clinical diagnostics.


Assuntos
Sondas de DNA/química , Membrana Eritrocítica/química , Corantes Fluorescentes/química , MicroRNAs/sangue , Animais , Carbocianinas/química , Sondas de DNA/genética , Ouro/química , Humanos , Limite de Detecção , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , Masculino , Camundongos Endogâmicos BALB C , MicroRNAs/genética , Nanoestruturas/química , Hibridização de Ácido Nucleico , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/genética , Espectrometria de Fluorescência/métodos
19.
Faraday Discuss ; 219(0): 138-153, 2019 10 30.
Artigo em Inglês | MEDLINE | ID: mdl-31313786

RESUMO

In the mucosal epithelium, the cellular glycocalyx can project tens to hundreds of nanometers into the extracellular space, erecting a physical barrier that provides protective functions, mediates the exchange of nutrients and regulates cellular interactions. Little is understood about how the physical properties of the mucosal glycocalyx influence molecular recognition at the cellular boundary. Here, we report the synthesis of PEG-based glycopolymers with tunable glycan composition, which approximate the extended architecture of mucin glycoproteins, and tether them to the plasma membranes of red blood cells (RBC) to construct an artificial mucin brush-like glycocalyx. We evaluated the association of two lectins, ConA and SNA, with their endogenous glycan ligands on the surface of the remodelled cells. The extended glycocalyx provided protection against agglutination of RBCs by both lectins; however, the rate and magnitude of ConA binding were attenuated to a greater degree in the presence of the glycopolymer spectators compared to those measured for SNA. The different sensitivity of ConA and SNA to glycocalyx crowding likely arises from the distinct presentation of their mannoside and sialoside receptors, respectively, within the native RBC glycocalyx.


Assuntos
Materiais Biomiméticos/metabolismo , Eritrócitos/metabolismo , Glicocálix/metabolismo , Hemaglutinação , Polietilenoglicóis/metabolismo , Materiais Biomiméticos/química , Concanavalina A/metabolismo , Membrana Eritrocítica/química , Membrana Eritrocítica/metabolismo , Eritrócitos/citologia , Glicocálix/química , Glicoconjugados/química , Glicoconjugados/metabolismo , Humanos , Mucinas/química , Mucinas/metabolismo , Lectinas de Plantas/metabolismo , Polietilenoglicóis/química , Polímeros/química , Polímeros/metabolismo , Proteínas Inativadoras de Ribossomos/metabolismo , Sambucus nigra/metabolismo
20.
Food Chem Toxicol ; 131: 110553, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31163221

RESUMO

Ginseng and its active gradient, ginsenoside Rg3 (Rg3), are widely used for a variety of health benefits, but concerns over their misuses are increasing. Previously, it has been reported that Rg3 can cause hemolysis, but its health outcome remains unknown. Here, we demonstrated that Rg3 could promote the procoagulant activity of erythrocytes through the process of hemolysis, ultimately leading to increased thrombosis. In freshly isolated human erythrocytes, Rg3 caused pore formation and fragmentation of the erythrocyte membrane. Confocal microscopy observation and flow cytometric analysis revealed that remnant erythrocyte fragments after the exposure to Rg3 expressed phosphatidylserine (PS), which can promote blood coagulation through providing assembly sites for coagulation complexes. Rat in vivo experiments further confirmed that intravenous administration of Rg3 produced PS-bearing erythrocyte debris and increased thrombosis. Collectively, we demonstrated that Rg3 could induce the procoagulant activity of erythrocytes by generating PS-bearing erythrocyte debris through hemolysis, which might provoke thrombosis.


Assuntos
Coagulação Sanguínea/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Ginsenosídeos/efeitos adversos , Hemólise/efeitos dos fármacos , Trombose/induzido quimicamente , Animais , Membrana Eritrocítica/química , Membrana Eritrocítica/efeitos dos fármacos , Eritrócitos/metabolismo , Eritrócitos/patologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Masculino , Fosfatidilserinas/química , Ratos Sprague-Dawley
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA