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Acta Academiae Medicinae Sinicae ; (6): 257-263, 2023.
Article in Chinese | WPRIM | ID: wpr-981261


Objective To examine the antiplatelet effect of ticagrelor by microfluidic chip and flow cytometry under shear stress in vitro. Methods Microfluidic chip was used to examine the effect of ticagrelor on platelet aggregation at the shear rates of 300/s and 1500/s.We adopted the surface coverage of platelet aggregation to calculate the half inhibition rate of ticagrelor.The inhibitory effect of ticagrelor on ADP-induced platelet aggregation was verified by optical turbidimetry.Microfluidic chip was used to construct an in vitro vascular stenosis model,with which the platelet reactivity under high shear rate was determined.Furthermore,the effect of ticagrelor on the expression of fibrinogen receptor (PAC-1) and P-selectin (CD62P) on platelet membrane activated by high shear rate was analyzed by flow cytometry. Results At the shear rates of 300/s and 1500/s,ticagrelor inhibited platelet aggregation in a concentration-dependent manner,and the inhibition at 300/s was stronger than that at 1500/s (both P<0.001).Ticagrelor at a concentration ≥4 μmol/L almost completely inhibited platelet aggregation.The inhibition of ADP-induced platelet aggregation by ticagrelor was similar to the results under flow conditions and also in a concentration-dependent manner.Ticagrelor inhibited the expression of PAC-1 and CD62P. Conclusion We employed microfluidic chip to analyze platelet aggregation and flow cytometry to detect platelet activation,which can reveal the responses of different patients to ticagrelor.

Humans , Ticagrelor/pharmacology , Platelet Aggregation Inhibitors/pharmacology , Flow Cytometry/methods , Microfluidics , Platelet Aggregation
Journal of Experimental Hematology ; (6): 495-502, 2023.
Article in Chinese | WPRIM | ID: wpr-982086


OBJECTIVE@#To study the effect of gradient shear stress on platelet aggregation by microfluidic chip Technology.@*METHODS@#Microfluidic chip was used to simulate 80% fixed stenotic microchannel, and the hydrodynamic behavior of the stenotic microchannel model was analyzed by the finite element analysis module of sollidwork software. Microfluidic chip was used to analyze the adhesion and aggregation behavior of platelets in patients with different diseases, and flow cytometry was used to detect expression of the platelet activation marker CD62p. Aspirin, Tirofiban and protocatechuic acid were used to treat the blood, and the adhesion and aggregation of platelets were observed by fluorescence microscope.@*RESULTS@#The gradient fluid shear rate produced by the stenosis model of microfluidic chip could induce platelet aggregation, and the degree of platelet adhesion and aggregation increased with the increase of shear rate within a certain range of shear rate. The effect of platelet aggregation in patients with arterial thrombotic diseases were significantly higher than normal group (P<0.05), and the effect of platelet aggregation in patients with myelodysplastic disease was lower than normal group (P<0.05).@*CONCLUSION@#The microfluidic chip analysis technology can accurately analyze and evaluate the platelet adhesion and aggregation effects of various thrombotic diseases unde the environment of the shear rate, and is helpful for auxiliary diagnosis of clinical thrombotic diseases.

Humans , Microfluidics , Platelet Adhesiveness , Platelet Aggregation , Blood Platelets/metabolism , Platelet Aggregation Inhibitors/pharmacology , Platelet Activation/physiology , Thrombosis
China Journal of Chinese Materia Medica ; (24): 4907-4921, 2021.
Article in Chinese | WPRIM | ID: wpr-921627


Platelet function tests have been increasingly used to assist in the diagnosis of platelet disorders and prethrombotic state, monitoring of the efficacy of antiplatelet therapies, and personalized treatment. On the basis of light transmission aggregometry, new methods for platelet function test have been developed successively. At present, the research and development of platelet function detector is in its infancy in China. The active constituents of antiplatelet Chinese medicines can be classified into terpenoids, flavonoids, saponins, organic acids, lignans, diketones, volatile oils, and stilbenes. The results of dose-antiplatelet effect relationship of Chinese medicines and the active constituents showed that the effective concentration of the extracts or monomers of Chinese medicines was at micromolar level(μmol·L~(-1)), among which salvianolic acid B and ginkgolide K, ginkgolide B, and ginkgolide A had the strongest antiplatelet effect. These results suggest that the antiplatelet effect of Chinese medicine may be weaker than that of chemical drugs and biological products. Therefore, it is necessary to explore the structure-activity relationship of the active constituents in existing Chinese medicines and further improve their efficacy through structure modification. The antiplatelet effect of Chinese medicines and the constituents involves multiple pathways and multiple targets. These research results provide a reference for clinical application of them. However, there is still a lack of large-scale multi-center clinical trials to confirm the efficacy and safety of them. The regularity of the relationship between the structures of various constituents and their corresponding functions is still unknown and the relevant signal transduction pathways and structure-activity relationship need to be further studied. This paper summarized and analyzed the determination methods of platelet functions and the research results of antiplatelet Chinese medicines, which is of reference value for the research of effective and safe antiplatelet Chinese medicines.

Biological Products , China , Medicine, East Asian Traditional , Platelet Aggregation Inhibitors/pharmacology , Platelet Function Tests
Environmental Health and Preventive Medicine ; : 70-70, 2020.
Article in English | WPRIM | ID: wpr-880306


BACKGROUND@#Resveratrol has been shown to inhibit platelet aggregation. However, the mechanism for this action of resveratrol remains to be clarified. The purpose of this study was to elucidate the Ca@*METHODS@#Ca@*RESULTS@#Thapsigargin-induced Ca@*CONCLUSIONS@#The results suggest that resveratrol inhibits thrombin-induced platelet aggregation through decreasing Ca

Humans , Antioxidants/administration & dosage , Calcium/physiology , Platelet Aggregation/drug effects , Platelet Aggregation Inhibitors/pharmacology , Resveratrol/pharmacology , Signal Transduction/drug effects
Arch. cardiol. Méx ; 89(4): 324-329, Oct.-Dec. 2019. tab, graf
Article in English | LILACS | ID: biblio-1149090


abstract Objective: Oral antiplatelet drugs are a key to modern pharmacotherapy in cardiovascular atherothrombotic diseases. Clopidogrel (CLO) constitutes the main preventive treatment of atherothrombosis. However, a considerable inter-individual variation in CLO response has been documented, resulting in suboptimal therapy and an increased risk of recurrent adverse effects in some patients. The enzyme CYP2C19 has been reported to be the CYP isoform that activates CLO to its active metabolite. Several single nucleotide polymorphisms in the CYP2C19 gene have been identified as strong predictors of CLO-impaired pharmacological response. At least 16 variants have been associated with changes in CYP2C19 activity. Materials and Methods: The following research was composed of a total of 102 subjects with high cardiovascular risk in the northeast of Mexico, with a maintenance dose of 75 mg of CLO per day. The platelet reactivity was measured with VerifyNow P2Y12 assay, while the presence of CYP2C19*2 was identified by real-time polymerase chain reaction. Results: Patients were categorized by CYP2C19 metabolizer status based on *2 genotypes using the common consensus star allele nomenclature as normal metabolizer (G/G), intermediate metabolizer (G/A), and poor metabolizer (A/A), respectively. The phenotype frequency for CYP2C19*2 was 74.5% (G/G), 21.6% (G/A), and 3.9% (A/A). The subjects with the A allele presented ≥235 P2Y12 reaction unit levels, classifying them how poor metabolizer. The prevalence of reduced CLO effectiveness was associated with the presence of CYP2C19*2 polymorphism among Mexican patients. Conclusion: The presence of the CYP2C19*2 allele is related to resistance to the antiplatelet effect of CLO (p = 0.003).

Resumen Objetivo: Los antiplaquetarios orales son clave en la farmacoterapia moderna de las enfermedades aterotrombóticas cardiovasculares. Clopidogrel (CLO) constituye el principal tratamiento preventivo de aterotrombosis (AT). Sin embargo, se ha documentado una considerable variación interindividual en la respuesta a CLO, lo que da como resultado una terapia subóptima y mayor riesgo de efectos adversos en algunos pacientes. La enzima CYP2C19 es la isoforma CYP que activa CLO a su metabolito activo. Se han identificado varios polimorfismos de un solo nucleótido en el gen CYP2C19 como fuertes predictores de respuesta farmacológica alterada a CLO. Al menos 16 variantes se han asociado con cambios en la actividad de CYP2C19. Método: Se reclutaron un total de 102 sujetos con alto riesgo cardiovascular del noreste de México, con dosis de mantenimiento de 75 mg de CLO/día. La reactividad plaquetaria se midió con el ensayo Verify Now P2Y12, la presencia de CYP2C19*2 se identificó mediante polymerase chain reaction en tiempo real. Resultado: Los pacientes fueron clasificados por el estado metabolizador CYP2C19*2 utilizando nomenclatura consenso, como metabolizador normal (G/G), metabolizador intermedio (G/A) y metabolizador pobre (A/A), respectivamente. La frecuencia del fenotipo para CYP2C19*2 fue 74.5% (G/G), 21.6% (G/A) y 3.9% (A/A). Los sujetos con alelo A presentaron ≥235 niveles P2Y12 reaction unit, clasificándolos como metabolizadores deficientes. La prevalencia de eficacia reducida a CLO se asoció con la presencia del polimorfismo CYP2C19*2 en pacientes mexicanos. Conclusiones: La presencia del alelo CYP2C19*2 se relaciona con resistencia al efecto antiagregante plaquetario del CLO (p = 0.003).

Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Platelet Aggregation Inhibitors/administration & dosage , Cardiovascular Diseases/drug therapy , Cytochrome P-450 CYP2C19/genetics , Clopidogrel/administration & dosage , Drug Resistance/genetics , Platelet Aggregation Inhibitors/pharmacology , Cardiovascular Diseases/physiopathology , Risk Factors , Polymorphism, Single Nucleotide , Alleles , Clopidogrel/pharmacology , Mexico
Arq. bras. cardiol ; 113(3): 357-363, Sept. 2019. tab, graf
Article in English | LILACS | ID: biblio-1038556


Abstract Background: High platelet reactivity (HPR) during therapy with acetylsalicylic acid (ASA) is a poor prognostic factor in acute coronary syndromes (ACS). The prevalence of HPR during ACS is greater than that reported in stable diseases. However, it is unclear whether this prevalence of HPR is a transient phenomenon or a characteristic of this high-risk population. Objective: The main objective is to compare the effects of ASA on platelet function in the initial and late phases of ACS in a single population. Secondary objectives are: correlation between the tests between themselves and the relationship between the tests and the variation of the inflammatory markers (C-reactive protein and interleukin-6). Methods: Seventy patients with non-ST segment elevation (NSTE) ACS in use of 100-200 mg of ASA per day for at least 7 days were prospectively studied. Platelet function was assessed in the first 48 hours and subsequently after 3 months using four methods: VerifyNow™ (VFN), whole blood platelet aggregation (WBPA) with arachidonic acid (AA) and collagen as agonists, and platelet function analyzer (PFA). The level of statistical significance considered was < 0.05. Results: According to the more specific methods (WBPA with AA and VFN), the incidence of HPR was significantly higher in the early phase than in the late phase: WBPA with AA: 31% versus 13%, p = 0.015; VFN: 32% versus 16%, p = 0.049. The other methods tested, which were less specific for ASA, did not show significant differences between phases. The correlation between the methods was weak or moderate (r ranging from 0.3 to 0.5, p < 0.05), and there were no significant associations between HPR and inflammatory markers. Conclusion: The prevalence of HPR during AAS therapy, assessed by specific methods for cyclooxygenase 1 (COX-1), is higher during the acute phase than in the late phase of NSTE ACS.

Resumo Fundamento: A alta atividade plaquetária (AAP) durante a terapia com ácido acetilsalicílico (AAS) é fator de mau prognóstico nas síndromes coronarianas agudas (SCA). A prevalência de AAP durante a SCA é maior do que a relatada na doença estável. No entanto, não está claro se esta prevalência de AAP é um fenômeno transitório ou característica dessa população de alto risco. Objetivo: O objetivo principal é comparar, em uma mesma população, os efeitos do AAS sobre a função plaquetária nas fases inicial e tardia da SCA. Os objetivos secundários são: correlação entre os testes entre si e a relação entre os testes e a variação dos marcadores inflamatórios (proteína C reativa e interleucina-6). Métodos: Foram estudados prospectivamente 70 pacientes com SCA sem elevação de ST (SCSST) em uso de 100 a 200 mg de AAS por dia por pelo menos 7 dias. A função plaquetária foi avaliada nas primeiras 48 horas e 3 meses após por quatro métodos: VerifyNow™ (VFN), agregometria de sangue total (AST) com ácido araquidônico (AA) e colágeno como agonistas, e analisador de função plaquetária (PFA). O nível de significância estatístico considerado foi < 0,05. Resultados: A média de idade foi de 65 ±9,7 anos e 54% da população eram do sexo feminino. De acordo com os métodos mais específicos (AST com AA e VFN), a incidência de AAP foi significativamente maior na fase inicial, em relação à tardia: AST com AA 31% versus 13%, p = 0,015; VFN 32% versus 16%, p = 0,049. Os outros métodos testados, menos específicos para o AAS, não mostraram diferenças significativas entre as fases. A correlação entre os métodos foi fraca ou moderada (r variando de 0,3 a 0,5, p < 0,05), e não houve associações significativas entre AAP e marcadores inflamatórios. Conclusão: A prevalência de AAP durante a terapia com AAS, avaliada por métodos específicos para cicloxigenase 1 (COX-1), é maior durante a fase aguda do que na tardia da SCASST.

Humans , Male , Female , Middle Aged , Aged , Blood Platelets/drug effects , Platelet Aggregation Inhibitors/therapeutic use , Aspirin/therapeutic use , Acute Coronary Syndrome/drug therapy , Platelet Function Tests , Blood Platelet Disorders/drug therapy , Blood Platelets/metabolism , Platelet Aggregation Inhibitors/pharmacology , Aspirin/pharmacology , Prospective Studies , Risk Factors , Non-ST Elevated Myocardial Infarction/physiopathology
In. Consolim-Colombo, Fernanda M; Saraiva, José Francisco Kerr; Izar, Maria Cristina de Oliveira. Tratado de Cardiologia: SOCESP / Cardiology Treaty: SOCESP. São Paulo, Manole, 4ª; 2019. p.122-127.
Monography in Portuguese | LILACS | ID: biblio-1009501
Rev. bras. cir. cardiovasc ; 33(4): 330-338, July-Aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-958435


Abstract Objective: The aims of this study were to determine whether the detection of preoperative clopidogrel resistance in patients undergoing cardiac surgery while using clopidogrel could play a guiding role in the prediction of postoperative excessive bleeding, transfusion requirements, and risks and to provide clinically significant data. Methods: Two hundred and twenty-two patients [median age: 59.4 (38-83) years; 38 females] undergoing emergency and elective coronary artery bypass graft (CABG) surgeries in our clinic were evaluated prospectively. Patients with multiple systemic diseases, other than diabetes mellitus (DM) and hypertension (HT), were excluded. Patients receiving clopidogrel were also evaluated for clopidogrel resistance and grouped according to the results of this test. Assessments of platelet functions were performed by multiplate impedance aggregometry method and adenosine diphosphate test. Results: The use of postoperative fresh blood replacement and platelet transfusion was higher in patients receiving clopidogrel than in those not receiving it (P=0.001, P=0.018). DM, HT, myocardial infarction, and the number of presentation to the emergency room were significantly higher in patients receiving clopidogrel than in those not receiving it (P<0.05). No significant difference was determined between patients with and without clopidogrel resistance regarding the amount of bleeding during and after surgery, erythrocyte suspension and fresh-frozen plasma transfusion rates, preoperative troponin values, ejection fraction values, and length of hospital stays (P>0.05). Conclusion: We think that resistance studies in patients receiving clopidogrel before cardiac surgery are not efficient to predict bleeding and bleeding-related complications in patients undergoing emergency and elective CABG surgeries.

Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Drug Resistance , Platelet Aggregation Inhibitors/pharmacology , Coronary Artery Bypass/adverse effects , Postoperative Hemorrhage/etiology , Clopidogrel/pharmacology , Platelet Function Tests/methods , Reference Values , Blood Transfusion , Predictive Value of Tests , Prospective Studies , Risk Factors , Statistics, Nonparametric , Risk Assessment/methods , Postoperative Hemorrhage/diagnosis , Preoperative Period
Bol. latinoam. Caribe plantas med. aromát ; 17(1): 36-52, ene. 2018. ilus, graf, tab
Article in English | LILACS | ID: biblio-915054


A comparative study of antioxidant properties, platelet antiaggregation activity and transcriptional analysis of flavonoid biosynthesis genes were performed in Fragaria x ananassa, F. vesca and F. chiloensis subsp chiloensis f. chiloensis and f. patagonica. Furthermore, differences in flavonoid content were found by UHPLC-MS. The highest free radical scavenging activity by DPPH assay was observed in F. chiloensis f. chiloensis, meanwhile, F. vesca presented the highest antioxidant capacity by FRAP. Biosynthetic flavonoids- related transcripts were higher abundant in F. x ananassa and lower in F. vesca. Additionally, all strawberry extracts showed antiaggregant effect (1 mg mL-1), but F. vesca and F. chiloensis subsp. chiloensis f. patagonica were still active at lower concentration. This study suggests that platelet antiaggregation effect of different strawberries could be due to isoflavones and flavonoids precursors in addition to anthocyanins. Results could usefully to take decisions in future breeding programs to improve the content of healthy compounds in strawberry fruits.

Se realizó un estudio comparativo de propiedades antioxidantes, actividad de antiagregacion plaquetaria, análisis transcripcional de genes de biosíntesis de flavonoides y contenido de estos en Fragaria x ananassa, F. vesca and F. chiloensis subsp chiloensis f. chiloensis and f. patagonica. La mayor actividad removedora de radicales libres por DPPH se observó en F. chiloensis f. chiloensis, mientras F. vesca presentó la mayor capacidad antioxidante mediante FRAP. Transcritos relacionados con biosíntesis de flavonoides fueron mas abundantes en F. x ananassa y menores en F. vesca. Adicionalmente, todos los extractos de frutillas mostraron efectos antiagregante (1 mg mL-1), pero F. vesca and F. chiloensis subsp. chiloensis f. patagonica fueron activos a concentraciones menores. Este estudio sugiere que efectos de antiagregación plaquetaria en distintas frutillas podría deberse a isoflavonas y precursores de flavonoides además de antocianinas. Los resultados podrían ser útiles en programas de mejoramiento genético para mejorar el contenido de compuestos saludables en frutilla.

Plant Extracts/chemistry , Fragaria/chemistry , Antioxidants/chemistry , Flavonoids/analysis , Flavonoids/genetics , Flavonoids/metabolism , Platelet Aggregation Inhibitors/pharmacology , Plant Extracts/pharmacology , Gene Expression , Free Radical Scavengers , Chromatography, High Pressure Liquid/methods , Fluorescence Recovery After Photobleaching , Real-Time Polymerase Chain Reaction , Antioxidants/pharmacology
Braz. j. med. biol. res ; 50(1): e5660, 2017. tab, graf
Article in English | LILACS | ID: biblio-839238


Clopidogrel and aspirin are the most commonly used medications worldwide for dual antiplatelet therapy after percutaneous coronary intervention. However, clopidogrel hyporesponsiveness related to gene polymorphisms is a concern. Populations with higher degrees of genetic admixture may have increased prevalence of clopidogrel hyporesponsiveness. To assess this, we genotyped CYP2C19, ABCB1, and PON1 in 187 patients who underwent percutaneous coronary intervention. Race was self-defined by patients. We also performed light transmission aggregometry with adenosine diphosphate (ADP) and arachidonic acid during dual antiplatelet therapy. We found a significant difference for presence of the CYP2C19*2 polymorphism between white and non-white patients. Although 7% of patients had platelet resistance to clopidogrel, this did not correlate with any of the tested genetic polymorphisms. We did not find platelet resistance to aspirin in this cohort. Multivariate analysis showed that patients with PON1 and CYP2C19 polymorphisms had higher light transmission after ADP aggregometry than patients with native alleles. There was no preponderance of any race in patients with higher light transmission aggregometry. In brief, PON1 and CYP2C19 polymorphisms were associated with lower clopidogrel responsiveness in this sample. Despite differences in CYP2C19 polymorphisms across white and non-white patients, genetic admixture by itself was not able to identify clopidogrel hyporesponsiveness.

Humans , Male , Female , Middle Aged , Aspirin/pharmacology , Blood Platelets/drug effects , Coronary Artery Disease/drug therapy , Platelet Aggregation Inhibitors/pharmacology , Ticlopidine/analogs & derivatives , Alleles , Aryldialkylphosphatase/genetics , ATP Binding Cassette Transporter, Subfamily B/genetics , Coronary Artery Disease/genetics , Cytochrome P-450 CYP2C19/genetics , Drug Therapy, Combination , Genotype , Percutaneous Coronary Intervention , Polymorphism, Genetic , Prospective Studies , Ticlopidine/pharmacology
Pakistan Journal of Pharmaceutical Sciences. 2017; 30 (1): 217-221
in English | IMEMR | ID: emr-185762


The study aimed to investigate the impact of clopidogrel combined with proton pump inhibitors [PPI] pantoprazole treatment on the prognosis of patients with transient ischemic attack [TIA]. A total of 478 cases of TIA patients treated with clopidogrel were randomly assigned half to clopidogrel combined with pantoprazole treatment and the control groups [clopidogrel treatment alone] from January 2012 to January 2014. The platelet aggregation before and after treatment and cerebrovascular events incidence within 90 days were compared and analyzed. Multivariate analysis was used to estimate the incidence of cerebrovascular events within 90 days. The platelet aggregation rate before treatment was 73.2 +/- 6.1% in the treatment group, 74.1 +/- 8.8% in the control group. The platelet aggregation rate after treatment was 38.1 +/- 10.7% in the treatment group, 36.8 +/- 9.7% in the control group. The platelet aggregation before and after treatments between the two groups had not significant difference [P>0.05]. The incidence of cerebrovascular events within 90 days [11.7% in the treatment group, 9.6% in the control group] between the two groups had not significant difference [P>0.05]. Multivariate analysis showed that the incidence of cerebrovascular events within 90 day was associated with hypertension [P=0.008], diabetes [P=0.000], hyperlipidemia [P=0.002] and ABCD2 score >3 points [P=0.000]. Clopidogrel combined with pantoprazole treatment had no significant effect on the prognosis of TIA patients

Aged , Female , Humans , Male , Middle Aged , Ticlopidine/analogs & derivatives , Platelet Aggregation Inhibitors/pharmacology , Purinergic P2Y Receptor Antagonists/pharmacology , 2-Pyridinylmethylsulfinylbenzimidazoles/pharmacology , Drug Therapy, Combination , Disease Progression
São Paulo; s.n; s.n; 2016. 134 p. graf, ilus, tab.
Thesis in Portuguese | LILACS | ID: biblio-881509


A terapia antiagregante é comumente indicada na prevenção e tratamento de doenças cardiovasculares. A dupla antiagregação com clopidrogrel e ácido acetilsalicílico (AAS) tem sido frequentemente adotada em pacientes com Doença Arterial Coronariana (DAC), mas apresenta ineficácia em uma parcela significativa da população com genótipo de respondedores. Essa falha terapêutica nos leva a questionar se outros mecanismos moleculares podem estar influenciando na resposta a esses fármacos. Recentes estudos sugerem que pequenas sequências de RNA não codificantes denominadas microRNAs (miRNAs) podem estar fortemente relacionadas com resposta ao tratamento fármaco-terapêutico, controlando as proteínas envolvidas na farmacocinética e farmacodinâmica. Entretanto, os principais miRNAs que atuam na dinâmica da resposta medicamentosa ainda não foram bem definidos. O objetivo deste estudo foi avaliar o perfil de miRNAs no sangue total periférico, procurando melhor esclarecer os mecanismos envolvidos na resposta aos antiagregantes plaquetários AAS e clopidogrel. Para isso, selecionou-se pacientes com DAC, os quais apresentavam diferentes respostas à dupla terapia de antiagregação determinadas pelo teste de agregação plaquetária. Baseados nos fenótipos, os perfis de expressão de miRNAs foram comparados entre os valores da taxa de agregação categorizados em tercis (T) de resposta. O grupo T1 foi constituído de pacientes respondedores, o T2 de respondedores intermediários e o T3 de não respondedores. Os perfis de miRNAs foram obtidos após sequenciamento de última geração e os dados obtidos foram analisados pelo pacote Deseq2. Os resultados mostraram 18 miRNAs diferentemente expressos entre os dois tercis extremos. Dentre esses miRNAs, 10 deles apresentaram importantes alvos relacionados com vias de ativação e agregação plaquetária quando analisados pelo software Ingenuity®. Dos 10 miRNAs, 4 deles, os quais apresentaram-se menos expressos no sequenciamento, demonstraram os mesmos perfis de expressão quando analisados pela reação em cadeia pela polimerase quantitativa (qPCR): hsa-miR-423-3p, hsa-miR-744-5p, hsa-miR- 30a-5p e hsa-let-7g-5p. A partir das análises de predição de alvos, pôde-se observar que os quatro miRNAs, quando menos expressos simultaneamente, predizem ativação da agregação plaquetária. Além disso, os miRNAs hsa-miR- 423-5p, hsa-miR-744-5p e hsa-let-7g-5p mostraram correlação com o perfil lipídico dos pacientes que, por sua vez, apresentou influência nos valores de agregação compreendidos no T3 de resposta a ambos os medicamentos. Sendo assim, conclui-se que maiores taxas de agregação plaquetária podem estar indiretamente relacionadas com os padrões de expressão de hsa-miR- 423-3p, hsa-miR-744-5p e hsa-let-7g-5p. Sugere-se que a avaliação do perfil de expressão destes 3 miRNAs no sangue periférico de pacientes com DAC possa predizer resposta terapêutica inadequada ao AAS e ao clopidogrel

The antiplatelet therapy is often indicated for the prevention and treatment of cardiovascular diseases. Dual antiplatelet therapy with clopidogrel and acetylsalicylic acid (ASA) has often been adopted in patients with coronary artery disease (CAD), but it has been ineffective in a significant portion of the population with genotype of responders. This fact leads us to question whether other molecular mechanisms may be influencing the response to these drugs. Recent studies suggest that small non-coding RNA sequences known as microRNAs (miRNAs) may be closely related to response to drug-therapeutic treatment, controlling proteins involved in pharmacokinetics and pharmacodynamics. The aim of this study was to evaluate the profile of miRNAs in whole blood, looking to better clarify mechanisms involved in ASA and clopidogrel response. For this purpose, we selected CAD patients who showed different responses to dual antiplatelet therapy determined by aggregation test. Based on the phenotypes, the miRNA expression profiles were compared between the platelet aggregation values categorized into tertiles (T) of response. The T1 group consisted of responding patients, the T2 consisted of intermediate responders and the T3 consisted of non-responders. The miRNA profiles were obtained after next-generation sequencing and data were analyzed by Deseq2 package. Results showed that 18 miRNAs were differentially expressed between the two extreme tertiles. By Ingenuity® software prediction analysis, 10 miRNAs showed important targets related with activation and aggregation of blood platelets. Of the 10 miRNAs, 4 of them, which were down-expressed on sequencing, showed the same fold-regulation when expression profiles were analyzed by quantitative polymerase chain reaction (qPCR): hsa-miR-423-3p, hsa-miR-744-5p, hsa-miR-30a-5p and has-hsa- let-7g-5p. By target prediction analysis, it was observed that, when the four miRNAs are simultaneously down-expressed, they predict activation of platelet aggregation. Furthermore, hsa-miR-423-5p, hsa-miR-744-5p, and hsa-let-7g-5p showed correlation with the lipid profile of patients which, in turn, demonstrated influence in aggregation values reaching T3 of response to both drugs. Therefore, we concluded that increased platelet aggregation rates may be indirectly related to the expression profiles of hsa-miR-423-3p, hsa-miR-744-5p and hsa-let-7g-5p. It is suggested that the evaluation of the expression profile of these three miRNAs in the peripheral blood of patients with CAD may predict inadequate therapeutic response to aspirin and clopidogrel

Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Coronary Artery Disease/pathology , Platelet Aggregation Inhibitors/pharmacology , Aspirin/pharmacology , MicroRNAs/analysis , Gene Library , Polymerase Chain Reaction/methods , Biological Specimen Banks
Braz. j. med. biol. res ; 48(6): 528-536, 06/2015. tab, graf
Article in English | LILACS | ID: lil-748220


This study aimed to determine the role of mitochondrial adenosine triphosphate-sensitive potassium (mitoKATP) channels and protein kinase C (PKC)-ε in the delayed protective effects of sevoflurane preconditioning using Langendorff isolated heart perfusion models. Fifty-four isolated perfused rat hearts were randomly divided into 6 groups (n=9). The rats were exposed for 60 min to 2.5% sevoflurane (the second window of protection group, SWOP group) or 33% oxygen inhalation (I/R group) 24 h before coronary occlusion. The control group (CON) and the sevoflurane group (SEVO) group were exposed to 33% oxygen and 2.5% sevoflurane for 60 min, respectively, without coronary occlusion. The mitoKATP channel inhibitor 5-hydroxydecanoate (5-HD) was given 30 min before sevoflurane preconditioning (5-HD+SWOP group). Cardiac function indices, infarct sizes, serum cardiac troponin I (cTnI) concentrations, and the expression levels of phosphorylated PKC-ε (p-PKC-ε) and caspase-8 were measured. Cardiac function was unchanged, p-PKC-ε expression was upregulated, caspase-8 expression was downregulated, cTnI concentrations were decreased, and the infarcts were significantly smaller (P<0.05) in the SWOP group compared with the I/R group. Cardiac function was worse, p-PKC-ε expression was downregulated, caspase-8 expression was upregulated, cTnI concentration was increased and infarcts were larger in the 5-HD+SWOP group (P<0.05) compared with the SWOP group. The results suggest that mitoKATP channels are involved in the myocardial protective effects of sevoflurane in preconditioning against I/R injury, by regulating PKC-ε phosphorylation before ischemia, and by downregulating caspase-8 during reperfusion.

Animals , Male , Ischemic Preconditioning, Myocardial/methods , Methyl Ethers/pharmacology , Myocardial Reperfusion Injury/prevention & control , Platelet Aggregation Inhibitors/pharmacology , Potassium Channels/pharmacology , Protein Kinase C/pharmacology , Anti-Arrhythmia Agents/pharmacology , Blotting, Western , /analysis , Decanoic Acids/pharmacology , Heart/drug effects , Heart/physiopathology , Hemodynamics/drug effects , Hydroxy Acids/pharmacology , Ischemia/prevention & control , Protective Agents/pharmacology , Random Allocation , Rats, Sprague-Dawley , Reproducibility of Results , Time Factors , Troponin I/analysis
Indian J Exp Biol ; 2015 Apr; 53(4): 228-231
Article in English | IMSEAR | ID: sea-158428


Sclerotiorin, isolated from the fermented broth of Penicillium frequentans, exhibited potent inhibition against human polymorphonuclear leukocytes 5-lipoxygenase and human platelet aggregation with a half maximal value 36 µM and 250 µM, respectively. Further, the Ames test has demonstrated the sclerotiorin to be non-mutagenic.

Arachidonate 5-Lipoxygenase/drug effects , Benzopyrans/pharmacology , Mutagenicity Tests , Neutrophils/enzymology , Penicillium/metabolism , Platelet Aggregation Inhibitors/pharmacology , Salmonella typhimurium/genetics
Acta cir. bras ; 30(2): 134-142, 02/2015. graf
Article in English | LILACS | ID: lil-741023


PURPOSE: To investigate the changes induced by DisBa-01 on repair of wound healing after induced incisional hernia (IH) in rats. METHODS: Thirty two male albino rats were submitted to IH and divided into four experimental groups: G1, placebo control; G2, DisBa-01-treated; G3, anti-αvβ3 antibodies-treated and G4, anti-α2 antibodies-treated. Histological, biochemical and extracellular matrix remodeling analysis of abdominal wall were evaluated. RESULTS: After 14 days, 100% of the G2 did not present hernia, and the hernia ring was closed by a thin membrane. In contrast, all groups maintained incisional hernia. DisBa-01 also increased the number macrophages and fibroblasts and induced the formation of new vessels. Additionally, MMP-2 was strongly activated only in G2 (p<0.05). Anti- αvβ3-integrin antibodies produced similar results than DisBa-01 but not anti-α2 integrin blocking antibodies. CONCLUSION: DisBa-01 has an important role in the control of wound healing and the blocking of this integrin may be an interesting therapeutically strategy in incisional hernia. .

Animals , Male , Disintegrins/pharmacology , Hernia, Ventral/pathology , /antagonists & inhibitors , Platelet Aggregation Inhibitors/pharmacology , Wound Healing/drug effects , Abdominal Wall/pathology , Collagen/analysis , Collagen/drug effects , Disease Models, Animal , Fibroblasts/drug effects , Hernia, Ventral/drug therapy , Hernia, Ventral/surgery , /analysis , /physiology , Random Allocation , Rats, Wistar , Reproducibility of Results , Time Factors , Treatment Outcome
Rev. panam. salud pública ; 36(5): 331-335, nov. 2014.
Article in Spanish | LILACS | ID: lil-733236


Desde diciembre de 2013, la Región de las Américas se enfrenta por primera vez a una epidemia de chikungunya. Los casos iniciales se registraron en el Caribe francés y, debido al comercio y la movilización de personas, esta epidemia no tardó en llegar a la República Dominicana, cuya población es de 10 millones de habitantes y comparte con Haití la isla La Española. En este artículo se difunde información extraída de diversos artículos y documentos oficiales sobre el virus, la infección y la epidemia de chikungunya, que han sido de gran ayuda para orientar la respuesta en la República Dominicana y pueden ser útiles para mejorar tanto el conocimiento como las actuaciones frente a la epidemia de los trabajadores del sector salud de la Región. Se destaca la importancia que revisten las investigaciones realizadas en países y territorios afectados del océano Índico, como la isla de Reunión, durante la epidemia declarada entre 2005 y 2007, cuando se registró una tasa de ataque mayor de 30%, se identificaron los grupos de riesgo, las formas graves y atípicas de la infección, la transmisión vertical del virus, las formas crónicas, que pueden provocar dolores recurrentes durante tres años, y las defunciones directa o indirectamente relacionadas con el virus chikungunya. Por su alta tasa de ataque, el virus chikungunya se convierte en un reto sin precedentes para los ministerios de salud, que exige una adecuada organización de los servicios de salud, la priorización de la atención a los grupos de riesgo y a los pacientes con formas graves de la enfermedad, así como una adecuada comunicación social y respuesta intersectorial.

The Region of the Americas has been affected since December 2013 by a chikungunya epidemic for the first time. Although the first cases were recorded in the French Caribbean, the epidemic quickly spread to the Dominican Republic due to trade and people movements. The Dominican Republic, which shares the island of Hispaniola with Haiti, has a population of 10 million. This article contains information from a range of different publications and official documents about the chikungunya virus infection and epidemic. These papers were extremely helpful for guiding the response to the epidemic in the Dominican Republic and may also be useful for enhancing knowledge of the virus and responses among health workers elsewhere in the region. Particular attention is drawn to the important research undertaken in countries and territories affected by the epidemic in the Indian Ocean area. This is the case, for example, of the island of La Réunion, where the epidemic had an attack rate of more than 30% between 2005 and 2007. Researchers were able to identify risk groups, severe and atypical forms of the infection, cases of vertical transmission, chronic disease causing recurrent pain over three years, and directly- or indirectly-related deaths from the virus. Given its high attack rate, the chikungunya virus has emerged as an exceptional challenge for health ministries and calls for appropriate organized responses from the health services, prioritization of care for risk groups and patients exhibiting severe forms of the disease, and effective social communication and intersectoral actions.

Animals , Rats , DNA , Angiotensin II/pharmacology , Muscle, Smooth, Vascular/drug effects , Platelet Aggregation Inhibitors/pharmacology , /analogs & derivatives , Vasoconstrictor Agents/pharmacology , Antihypertensive Agents/pharmacology , Benzimidazoles/pharmacology , Biphenyl Compounds/pharmacology , Cell Division/physiology , Cells, Cultured , Muscle, Smooth, Vascular/physiology , Proto-Oncogene Proteins c-fos/biosynthesis , RNA, Messenger/metabolism , Rats, Inbred WKY , Tetrazoles/pharmacology , /pharmacology
Einstein (Säo Paulo) ; 12(3): 366-373, Jul-Sep/2014.
Article in English | LILACS | ID: lil-723923


Personalized medicine is the use of biomarkers, most of them molecular markers, for detection of specific genetic traits to guide various approaches for preventing and treating different conditions. The identification of several genes related to heredity, oncology and infectious diseases lead to the detection of genetic polymorphisms that are involved not only in different clinical progression of these diseases but also in variations in treatment response. Currently, it is possible to detect these polymorphisms using several methodologies: detection of single nucleotide polymorphisms using polymerase chain reaction methods; nucleic acid microarray detection; and nucleic acid sequencing with automatized DNA sequencers using Sanger-derived methods and new generation sequencing. Personalized medicine assays are directed towards detecting genetic variations that alter interactions of drugs with targets or the metabolic pathways of drugs (upstream and downstream) and can be utilized for the selection of drug formulations and detect different immunogenicities of the drug. Personalized medicine applications have already been described in different areas of Medicine and allow specific treatment approaches to be applied to each patient and pathology according to the results of these assays. The application of such a protocol demands an increasing interaction between the clinical laboratory and the clinical staff. For its implementation, a coordinated team composed of basic researchers and physicians highly specialized in their areas supported by a highly specialized team of clinical analysts particularly trained in molecular biology assays is necessary.

Medicina personalizada é o uso de biomarcadores, em sua maioria marcadores moleculares, para a detecção de traços genéticos específicos, a fim de orientar diversas abordagens para a prevenção e o tratamento de diferentes doenças. A identificação de vários genes relacionados a doenças hereditárias, oncológicas e infecciosas permite a detecção de polimorfismos genéticos que estão envolvidos em diferentes evoluções clínicas dessas doenças, bem como com variações na resposta ao tratamento. Atualmente, já é possível detectar esses polimorfismos utilizando diversas metodologias: a detecção de polimorfismos de nucleotídeo único pela reação de polimerização em cadeia; a detecção de microarranjos de ácidos nucleicos; e o sequenciamento de ácidos nucleicos com sequenciadores de DNA automatizados usando métodos derivados de sequenciamento Sanger ou de nova geração. Os ensaios de medicina personalizada são dirigidos para detectar variações genéticas que alteram interações de fármacos com alvos ou vias metabólicas de fármacos (anabólicas e catabólicas), podendo ser utilizados para a seleção de formulações farmacêuticas e para detectar diferentes imunogenicidades da droga. As aplicações de medicina personalizada já foram descritas em várias áreas da Medicina e permitem que abordagens de tratamento específicas sejam aplicadas para cada paciente e para cada doença, de acordo com os resultados dos ensaios utilizados. A aplicação de um protocolo desse tipo exige uma relação intensa entre o laboratório e o corpo clínico. Para sua execução, é necessária uma equipe coordenada, composta por investigadores de pesquisa básica e médicos altamente especializados em suas áreas, apoiada por um time bastante especializado de analistas clínicos treinados em testes de biologia molecular.

Humans , Genetic Markers/genetics , Precision Medicine/methods , Neoplasms/drug therapy , Neoplasms/genetics , Anticoagulants/pharmacology , Hepatitis C/drug therapy , Hepatitis C/genetics , Pharmacogenetics , Platelet Aggregation Inhibitors/pharmacology , Biomarkers, Tumor/genetics
Acta cir. bras ; 29(supl.2): 34-37, 2014. graf
Article in English | LILACS | ID: lil-721385


PURPOSE: To investigate the blood vessels' concentration in TRAM flap's rat model, in the presence of pentoxifylline. METHODS: 32 male, Wistar-EPM rats were divided into two groups. Control group (C): 0.5 ml of saline, intraperitoneally, once a day, for seven days before flap elevation; PTX group (P): pentoxifylline (20mg/kg/day), intraperitoneally, for seven days before flap elevation. After that, they were submitted to a caudal unipedicle TRAM flap. On the fifth postoperative day, percentages of flap necrosis were determined via the "paper template" method and Tram flap's zone IV skin biopsies were taken for histological analysis. RESULTS: the mean percentage of flap necrosis in group C was 58.7 % and in group P, 31.1 (Wilcoxon test; p = 0.003). Mean capillary vessels number in zone IV's skin in C group was 33.4 and in P group was 71.9 (p=0.008). CONCLUSIONS: Pentoxifylline was effective reducing the necrosis in the caudal unipedicle TRAM flap in the rat as well as increasing the number of capillaries in an ischemic zone (zone IV). .

Animals , Male , Capillaries/drug effects , Pentoxifylline/pharmacology , Platelet Aggregation Inhibitors/pharmacology , Rectus Abdominis/drug effects , Surgical Flaps/blood supply , Tissue Survival/drug effects , Biopsy , Capillaries/pathology , Models, Animal , Necrosis , Random Allocation , Rats, Wistar , Reproducibility of Results , Rectus Abdominis/blood supply , Rectus Abdominis/pathology , Skin/blood supply , Skin/drug effects , Skin/pathology , Surgical Flaps/pathology
Arq. bras. cardiol ; 101(3): 277-282, set. 2013. ilus, tab
Article in Portuguese | LILACS | ID: lil-686535


As plaquetas estão envolvidas em vários processos biológicos, desde o combate a agentes infecciosos até a coordenação do controle da permeabilidade vascular e angiogênese. Entretanto, o seu principal foco de ação consiste na modulação da cascata de coagulação. A intervenção coronariana percutânea é um procedimento com alto risco trombogênico, que induz a ativação plaquetária e de monócitos, devido à lesão direta do endotélio e pelo contato de estruturas trombogênicas com o sangue, levando ao aumento da atividade inflamatória, tanto no local do dano vascular coronariano como de forma sistêmica. Os receptores plaquetários P2Y12 desempenham papel central na amplificação da agregação induzida por todos os agonistas plaquetários, como a adenosina difosfato, o colágeno, tromboxano A2, adrenalina e serotonina. Por esse motivo, têm sido o principal alvo das drogas antiplaquetárias. Apesar de atuarem no mesmo receptor, características farmacocinéticas e farmacodinâmicas distintas conferem peculiaridades a cada agente.

Apart from their role in hemostasis and thrombosis, platelets are involved in many other biological processes such as wound healing and angiogenesis. Percutaneous coronary intervention is a highly thrombogenic procedure inducing platelets and monocytes activation through endothelial trauma and contact activation by intravascular devices. Platelet P2Y12 receptor activation by adenosine diphosphate facilitates non-ADP agonist-mediated platelet aggregation, dense granule secretion, procoagulant activity, and the phosphorylation of several intraplatelet proteins, making it an ideal drug target. However, not all compounds that target the P2Y12 receptor have similar efficacy and safety profiles. Despite targeting the same receptor, the unique pharmacologic properties of each of these P2Y12 receptor-directed compounds can lead to very different clinical effects.

Humans , Percutaneous Coronary Intervention , Platelet Aggregation Inhibitors/pharmacology , Platelet Aggregation/drug effects , /pharmacology , /drug effects , Adenosine Monophosphate/analogs & derivatives , Adenosine Monophosphate/pharmacology , Adenosine/analogs & derivatives , Adenosine/pharmacology , Piperazines/pharmacology , Thienopyridines/pharmacology , Thiophenes/pharmacology