scholarly journals Increased Pro-Thrombotic Platelet Activity Associated with Thrombin/PAR1-Dependent Pathway Disorder in Patients with Secondary Progressive Multiple Sclerosis

2020 ◽  
Vol 21 (20) ◽  
pp. 7722
Author(s):  
Angela Dziedzic ◽  
Elzbieta Miller ◽  
Michal Bijak ◽  
Lukasz Przyslo ◽  
Joanna Saluk-Bijak
Keyword(s):  
Multiple Sclerosis ◽  
Mrna Expression ◽  
Platelet Activation ◽  
Blood Platelets ◽  
Surface Expression ◽  
Epidemiological Studies ◽  
Progressive Multiple Sclerosis ◽  
Apolipoprotein A1 ◽  
Platelet Activity ◽  
Activation Markers

Epidemiological studies confirm the high risk of ischemic events in multiple sclerosis (MS) that are associated with increased pro-thrombotic activity of blood platelets. The most potent physiological platelet agonist is thrombin, which activates platelets via cleavage of specific protease-activated receptors (PARs). Our current study is aimed to determine the potential genetics and proteomic abnormalities of PAR1 in both platelets and megakaryocytes, which may have thromboembolic consequences in the course of MS. The obtained results were correlated with the expression level of platelet and megakaryocyte transcripts for APOA1 and A2M genes encoding atherosclerosis biomarkers: apolipoprotein A1 (ApoA1) and α-2-macroglobulin (α2M), respectively. Moreover, PAR1 functionality in MS platelets was assessed by flow cytometry, determining the level of platelet–platelet and platelet–leukocyte aggregates, platelet microparticles and surface expression of P-selectin. As a PAR1 agonist, the synthetic TRAP-6 peptide was used, which made it possible to achieve platelet activation in whole blood without triggering clotting. Comparative analyses showed an elevated level of platelet activation markers in the blood of MS patients compared to controls. The mRNA expression of gene coding α2M was upregulated, whilst ApoA1 was down-regulated, both in platelets and megakaryocytes from MS patients. Furthermore, we observed an increase in both mRNA expression and surface density of PAR1 in platelets and megakaryocytes in MS compared to controls. Both the level of platelet activation markers and PAR1 expression showed a high correlation with the expression of transcripts for APOA1 and A2M genes.

scholarly journals Oxidative Damage of Blood Platelets Correlates with the Degree of Psychophysical Disability in Secondary Progressive Multiple Sclerosis

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Angela Dziedzic ◽  
Agnieszka Morel ◽  
Elzbieta Miller ◽  
Michal Bijak ◽  
Tomasz Sliwinski ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Membrane Potential ◽  
Mrna Expression ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Blood Platelets ◽  
Fold Increase ◽  
Past Research ◽  
Progressive Multiple Sclerosis ◽  
Nitrative Stress

The results of past research studies show that platelets are one of the main sources of reactive oxygen species (ROS) and reactive nitrogen species (RNS) to be found in the course of many pathological states. The aim of this study was to determine the level of oxidative/nitrative stress biomarkers in blood platelets obtained from multiple sclerosis (MS) patients (n=110) and to verify their correlation with the clinical parameters of the psychophysical disability of patients. The mitochondrial metabolism of platelets was assessed by measuring the intracellular production of ROS using the fluorescence method with DCFH-DA dye and by identification of changes in the mitochondrial membrane potential of platelets using the JC-1 dye. Moreover, we measured the mRNA expression for the gene encoding the cytochrome c oxidase subunit I (MTCO-1) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in platelets and megakaryocytes using the RT-qPCR method, as well as the concentration of NADPH oxidase (NOX-1) by the ELISA method. Our results proved an increased level of oxidative/nitrative damage of proteins (carbonyl groups, 3-nitrotyrosine) (p<0.0001) and decreased level of -SH in MS (p<0.0001) and also a pronounced correlation between these biomarkers and parameters assessed by the Expanded Disability Status Scale and the Beck’s Depression Inventory. The application of fluorescence methods showed mitochondrial membrane potential disruption (p<0.001) and higher production of ROS in platelets from MS compared to control (p<0.0001). Our research has also confirmed the impairment of red-ox metabolism in MS, which was achieved by increasing the relative mRNA expression in platelets for the genes studied (2-fold increase for the MTCO-1 gene and 1.5-fold increase in GAPDH gene, p<0.05), as well as the augmented concentration of NOX-1 compared to control (p<0.0001). Our results indicate that the oxidative/nitrative damage of platelets is implicated in the pathophysiology of MS, which reflects the status of the disease.

scholarly journals Pro-Thrombotic Activity of Blood Platelets in Multiple Sclerosis

Cells ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 110 ◽  
Author(s):  
Joanna Saluk-Bijak ◽  
Angela Dziedzic ◽  
Michal Bijak
Keyword(s):  
Multiple Sclerosis ◽  
Platelet Activation ◽  
Inflammatory Diseases ◽  
Blood Platelets ◽  
Experimental Studies ◽  
Epidemiological Studies ◽  
Membrane Receptors ◽  
Vascular Integrity ◽  
Oxidative Processes ◽  
Specific Membrane

The available data, including experimental studies, clearly indicate an excessive intravascular activation of circulating platelets in multiple sclerosis (MS) and their hyper-responsiveness to a variety of physiological activators. Platelet activation is manifested as an increased adhesion and aggregation and is accompanied by the formation of pro-thrombotic microparticles. Activated blood platelets also show an expression of specific membrane receptors, synthesis many of biomediators, and generation of reactive oxygen species. Epidemiological studies confirm the high risk of stroke or myocardial infarction in MS that are ischemic incidents, strictly associated with incorrect platelet functions and their over pro-thrombotic activity. Chronic inflammation and high activity of pro-oxidative processes in the course of MS are the main factors identified as the cause of excessive platelet activation. The primary biological function of platelets is to support vascular integrity, but the importance of platelets in inflammatory diseases is also well documented. The pro-thrombotic activity of platelets and their inflammatory properties play a part in the pathophysiology of MS. The analysis of platelet function capability in MS could provide useful information for studying the pathogenesis of this disease. Due to the complexity of pathological processes in MS, medication must be multifaceted and blood platelets can probably be identified as new targets for therapy in the future.

Anthocyanins from red cabbage extract — evidence of protective effects on blood platelets

2012 ◽  
Vol 7 (4) ◽  
pp. 655-663 ◽  
Author(s):  
Joanna Saluk ◽  
Michał Bijak ◽  
Joanna Kołodziejczyk-Czepas ◽  
Małgorzata Posmyk ◽  
Krystyna Janas ◽  
...  
Keyword(s):  
Cardiovascular Diseases ◽  
Platelet Activation ◽  
Blood Platelets ◽  
Protective Effects ◽  
Platelet Activity ◽  
Red Cabbage ◽  
Arachidonate Cascade ◽  
Wide Range ◽  
Antioxidative Effects

AbstractRed cabbage belongs to cruciferous vegetables recognized as a rich source of anthocyanins. Anthocyanins have a wide range of therapeutic advantages without adverse effects, including cardiovascular protective properties. For development of cardiovascular diseases, platelet activation is crucial; therefore compounds which inhibit platelet activation are sought after. The anti-platelet activity of anthocyanins has only been described and is still unclear. In our study, the extract of anthocyanins, obtained from fresh leaves of red cabbage, was used in vitro to examine their antioxidative effects on platelets under oxidative stress conditions which are responsible for hyperactivity of these cells. The antiplatelet and antioxidative activities were determined by platelet aggregation and specific markers of the arachidonate cascade with O2−· generation, and oxidative changes (carbonyl groups and 3-nitrotyrosine). Extracts (5–15 μM) protected platelet proteins and lipids against oxidative damage, and diminished platelet activation. Anthocyanins from red cabbage provided beneficial anti-platelet effects and might help prevent cardiovascular diseases.

Cigarette smoking and urinary excretion of markers for platelet/vessel wall interaction in healthy women

1991 ◽  
Vol 81 (1) ◽  
pp. 11-15 ◽  
Author(s):  
C. Rångemark ◽  
Å. Wennmalm
Keyword(s):  
Cigarette Smoking ◽  
Urinary Excretion ◽  
Platelet Activation ◽  
Vessel Wall ◽  
Thromboxane A2 ◽  
Epidemiological Studies ◽  
Thromboxane B2 ◽  
Platelet Activity ◽  
Healthy Women ◽  
Wall Interaction

1. Cigarette smoking is known to increase the risk of cardiovascular disease in both men and women. Experimental and epidemiological studies have demonstrated that cigarette smoking is associated with several indices of increased platelet activation and platelet/vessel wall interaction in men. The aim of the present study was to test the hypothesis that cigarette smoking is linked to an increased platelet activity in women also. 2. In 26 healthy smoking and non-smoking women (age 21–49 years) the urinary excretion of the thromboxane A2 metabolite 2,3-dinor-thromboxane B2 (an index of platelet activation) and of the prostacyclin metabolite 2,3-dinor-6-keto-prostaglandin F1α (an index of platelet/vessel wall interaction) were analysed by g.c.-m.s. in samples collected on days 3, 10 and 20 of their respective menstrual cycles. 3. The urinary excretion of 2,3-dinor-thromboxane B2 did not vary significantly during the menstrual cycle, either in the smokers or in the non-smokers. It was consistently higher (P < 0.004) in the group of smokers (average of days 3, 10 and 20, 395 ± 61 pg/mg of creatinine; mean ± sem) than in the group of non-smokers (average 188 ± 22 pg/mg of creatinine). 4. The urinary excretion of 2,3-dinor-6-keto-prostaglandin F1α did not differ between the groups on any of the days studied (average on days 3, 10 and 20 in the smokers and non-smokers was 281 ± 50 and 227 ± 30 pg/mg of creatinine, respectively). 5. These data demonstrate that smoking fertile women excrete more of the thromboxane A2 metabolite than do non-smokers, thereby supporting the hypothesis that cigarette smoking elicits platelet activation in healthy women. In contrast, platelet/vessel wall interaction does not appear to be facilitated in smoking compared with non-smoking women. The data suggest that platelet activation is not a major haemostatic mechanism during menstruation.

scholarly journals Sample Preparation as a Critical Aspect of Blood Platelet Mitochondrial Respiration Measurements—The Impact of Platelet Activation on Mitochondrial Respiration

2021 ◽  
Vol 22 (17) ◽  
pp. 9332
Author(s):  
Karolina Siewiera ◽  
Magdalena Labieniec-Watala ◽  
Nina Wolska ◽  
Hassan Kassassir ◽  
Cezary Watala
Keyword(s):  
Platelet Activation ◽  
Mitochondrial Respiration ◽  
Blood Platelets ◽  
High Sensitivity ◽  
Blood Platelet ◽  
High Electron ◽  
Activation Markers ◽  
The Media ◽  
The Impact ◽  
Electron Transfer Capacity

Blood platelets are considered as promising candidates as easily-accessible biomarkers of mitochondrial functioning. However, their high sensitivity to various stimulus types may potentially affect mitochondrial respiration and lead to artefactual outcomes. Therefore, it is crucial to identify the factors associated with platelet preparation that may lead to changes in mitochondrial respiration. A combination of flow cytometry and advanced respirometry was used to examine the effect of blood anticoagulants, the media used to suspend isolated platelets, respiration buffers, storage time and ADP stimulation on platelet activation and platelet mitochondria respiration. Our results clearly show that all the mentioned factors can affect platelet mitochondrial respiration. Briefly, (i) the use of EDTA as anticoagulant led to a significant increase in the dissipative component of respiration (LEAK), (ii) the use of plasma for the suspension of isolated platelets with MiR05 as a respiration buffer allows high electron transfer capacity and low platelet activation, and (iii) ADP stimulation increases physiological coupling respiration (ROUTINE). Significant associations were observed between platelet activation markers and mitochondrial respiration at different preparation steps; however, the fact that these relationships were not always apparent suggests that the method of platelet preparation may have a greater impact on mitochondrial respiration than the platelet activation itself.

scholarly journals The Changes of Blood Platelet Reactivity in the Presence of Crude Extracts Isolated from Leaves and Twigs of Elaeagnus Rhamnoides (L.) A. Nelson Measuring in Whole Blood

2021 ◽  
Author(s):  
Bartosz Skalski ◽  
Joanna Rywaniak ◽  
Jerzy Żuchowski ◽  
Anna Stochmal ◽  
Beata Olas
Keyword(s):  
Phenolic Compounds ◽  
Platelet Activation ◽  
Whole Blood ◽  
Leaf Extract ◽  
Platelet Reactivity ◽  
Blood Platelets ◽  
Blood Platelet ◽  
Surface Expression ◽  
Sea Buckthorn ◽  
Crude Extracts

Abstract Uncontrolled blood platelet activation is an important risk factor of cardiovascular disease (CVDs). Various studies on phenolic compounds indicate that they have a protective effect on the cardiovascular system through different mechanisms, including the reduction of blood platelet activation. One of the plants that is particularly rich in phenolic compounds is sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson). The aim of the present study in vitro was to determine the anti-platelet properties of crude extracts isolated from leaves and twigs of E. rhamnoides (L.) A. Nelson in whole blood using flow cytometric and total thrombus-formation analysis system (T-TAS). The aim of our study was also analyze of blood platelet proteoms in the presence of different sea buckthorn extracts. A significant new finding is a decrease surface expression of P-selectin on blood platelets stimulated by 10 µM ADP and 10 µg/mL collagen, and a decrease surface expression of GPIIb/IIIa active complex on non-activated platelets and platelets stimulated by 10 µM ADP and 10 µg/mL collagen in the presence of sea buckthorn leaf extract (especially at the concentration 50 µg/mL). The twig extract also displayed antiplatelet potential. However, this activity was higher in the leaf extract than in the twig extract in whole blood. In addition, our present findings clearly demonstrate that investigated plant extracts have anticoagulant properties (measured by T-TAS). Therefore, the two tested extracts may be promising candidates for the natural anti-platelet and anticoagulant supplements.

scholarly journals Preparations from Selected Cucurbit Vegetables as Antiplatelet Agents – An in Vitro Study

2021 ◽  
Author(s):  
Agata Rolnik ◽  
Bartosz Skalski ◽  
Anna Stochmal ◽  
Beata Olas
Keyword(s):  
Platelet Activation ◽  
Cucurbita Pepo ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
In Vitro Study ◽  
Blood Platelet ◽  
Arachidonic Acid Metabolism ◽  
Platelet Activity ◽  
Activated Platelets

Abstract Increased blood platelet activation plays an important role in cardiovascular diseases (CVDs). Recent experiments indicate that certain fruits and vegetables, including onion, garlic, and beetroot, have anti-platelet potential and therefore may reduce the likelihood of CVDs. While vegetables from the Cucuritaceae family are known to exerting beneficial antioxidant and anti-inflammatory effects, their effects on blood platelet activation are poorly understood. Therefore, the aim of the present study was to determine the effect on platelet adhesion of preparations from selected cucurbits: pumpkin (Cucirbita pepo; fruit without seeds), zucchini (Cucurbita pepo convar. giromontina; fruit with seeds), cucumber (Cucumis sativus; fruit with seeds), white pattypan squash (Cucurbita pepo var. patisoniana; fruit without seeds) and yellow pattypan squash (Cucurbita pepo var. patisoniana, fruit without seeds). It also evaluates the activity of these preparations on enzymatic lipid peroxidation in thrombin-activated washed blood platelets by TBARS assay. The study also determines the anti-platelet and anticoagulant properties of these five cucurbit preparations in whole blood by flow cytometry and with the total thrombus-formation analysis system (T-TAS) and evaluates the cytotoxicity of the tested preparations against platelets based on LDH activity. The results indicate that the yellow Cucurbita pepo var. patisoniana preparation demonstrated stronger anti-platelet properties than the other tested preparations, reducing the adhesion of thrombin-activated platelets to collagen/fibrinogen, and inhibiting arachidonic acid metabolism and GPIIb/IIIa expression on 10 µM ADP-activated platelets. None of the preparations was found to cause platelet lysis. Our findings provide new information on the anti-platelet activity of the tested cucurbit preparations and their potential for treating CVDs associated with platelet hyperactivity.

Abstract 53: Role of Serotonin through 5HT 2A in alphaIIbBeta3 Outside-in Activation

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Kendra H Oliver ◽  
Matthew Duvernay ◽  
Heidi E Hamm ◽  
Ana M Carneiro
Keyword(s):  
Platelet Activation ◽  
Selective Serotonin Reuptake Inhibitors ◽  
Critical Role ◽  
Surface Expression ◽  
Clear Understanding ◽  
Storage Site ◽  
Platelet Activity ◽  
Increased Risk ◽  
Receptor Surface Expression

Patients taking selective serotonin reuptake inhibitors (SSRIs) have an increased risk of bleeding. However it remains unclear how SSRIs, which inhibit the serotonin transporter (SERT), modulate hemostasis, particularly platelet activation. 5HT (serotonin, 5-hydroxytryptamine) is taken up by platelets via SERT and activates the 5HT 2A receptor. αIIbβ3 is an integrin that binds fibrinogen and plays a critical role in platelet aggregation, which is central to hemostasis and thrombosis. Futhermore, it is known that αIIbβ3 and SERT interact genetically, physically, and functionally. To gain a clear understanding of how SERT function regulates platelet activity we examined two independent models of SERT inhibition - constitutive genetic deletion (SERT -/- ) and 6-day water citalopram administration. Through SERT, platelets are the major storage site for 5HT in whole blood. Our works suggests that ADP-mediated αIIbβ3 activation is decreased as assessed by JON/a binding. Additionally while 5HT alone does not cause any platelet activation in SERT +/+ , 5HT synergizes with submaximal ADP to potentiate αIIbβ3 activation. The synergy of 5HT-ADP synergy is lost in SERT -/- platelets as determined by the addition of exogenous 5HT. Furthermore, blocking 5HT 2A receptor with an antagonist, ketanserin, inhibited ADP-mediated αIIbβ3 activation, suggesting that 5HT-ADP synergy could be mediated through activation of the 5HT 2A receptor. Additionally, we found that 5HT 2A receptor surface expression was reduced in SERT -/- platelets. SERT -/- platelet membranes incubated with a 5HT 2A agonist, DOI, also exhibited reduced 35S GTPys incorporation. These findings suggest that long-term inhibition of SERT leads to reduced 5HT 2A receptor-dependent potentiation of αIIbβ3 activation, perhaps through altered hemostatic regulation of 5HT. Further experiments are being carried out to determine a possible novel regulation of αIIbβ3 via 5HT 2A that could alter the functionality of αIIbβ3. However, these findings suggest a novel mechanism central to the increased bleeding observed in patients taking SSRIs and support the continued investigation of 5HT 2A receptor inhibitor as anti-platelet drugs for treatment of cardiovascular diseases.

scholarly journals Relationship between the Increased Haemostatic Properties of Blood Platelets and Oxidative Stress Level in Multiple Sclerosis Patients with the Secondary Progressive Stage

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Agnieszka Morel ◽  
Michał Bijak ◽  
Elżbieta Miller ◽  
Joanna Rywaniak ◽  
Sergiusz Miller ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Multiple Sclerosis ◽  
Early Stage ◽  
Blood Platelets ◽  
Control Group ◽  
Platelet Activity ◽  
Secondary Progressive ◽  
Primary Hemostasis ◽  
Scientific Papers ◽  
Multiple Sclerosis Patients

Multiple sclerosis (MS) is the autoimmune disease of the central nervous system with complex pathogenesis, different clinical courses and recurrent neurological relapses and/or progression. Despite various scientific papers that focused on early stage of MS, our study targets selective group of late stage secondary progressive MS patients. The presented work is concerned with the reactivity of blood platelets in primary hemostasis in SP MS patients. 50 SP MS patients and 50 healthy volunteers (never diagnosed with MS or other chronic diseases) were examined to evaluate the biological activity of blood platelets (adhesion, aggregation), especially their response to the most important physiological agonists (thrombin, ADP, and collagen) and the effect of oxidative stress on platelet activity. We found that the blood platelets from SP MS patients were significantly more sensitive to all used agonists in comparison with control group. Moreover, the platelet hemostatic function was advanced in patients suffering from SP MS and positively correlated with increased production ofO2-∙in these cells, as well as with Expanded Disability Status Scale. We postulate that the increased oxidative stress in blood platelets in SP MS may be primarily responsible for the altered haemostatic properties of blood platelets.

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