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 Anti-Platelet Properties of Phenolic Extracts from the Leaves and Twigs of Elaeagnus rhamnoides (L.) A. Nelson

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3620 ◽  
Author(s):  
Bartosz Skalski ◽  
Bogdan Kontek ◽  
Agata Rolnik ◽  
Beata Olas ◽  
Anna Stochmal ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Blood Platelets ◽  
Biological Effects ◽  
Blood Platelet ◽  
Arachidonic Acid Metabolism ◽  
Sea Buckthorn ◽  
Platelet Activity ◽  
Aronia Melanocarpa ◽  
Phenolic Extracts

Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) is a small tree or bush. It belongs to the Elaeagnaceae family, and has been used for many years in traditional medicine in both Europe and Asia. However, there is no data on the effect of sea buckthorn leaves and twigs on the properties of blood platelets. The aim of the study was to analyze the biological activity of phenolic extracts from leaves and twigs of sea buckthorn in blood platelets in vitro. Two sets of extracts were used: (1) phenolic compounds from twigs and (2) phenolic compounds from leaves. Their biological effects on human blood platelets were studied by blood platelet adhesion, platelet aggregation, arachidonic acid metabolism and the generation of superoxide anion. Cytotoxicity was also evaluated against platelets. The action of extracts from sea buckthorn twigs and leaves was compared to activities of the phenolic extract (a commercial product from the berries of Aronia melanocarpa (Aronox®) with antioxidative and antiplatelet properties. This study is the first to demonstrate that extracts from sea buckthorn leaves and twigs are a source of bioactive compounds which may be used for the prophylaxis and treatment of cardiovascular pathologies associated with blood platelet hyperactivity. Both leaf and twig extracts were found to display anti-platelet activity in vitro. Moreover, the twig extract (rich in proanthocyanidins) displayed better anti-platelet potential than the leaf extract or aronia extract.

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 In vivo effects of eltrombopag on platelet function in immune thrombocytopenia: no evidence of platelet activation

Blood ◽  
2012 ◽  
Vol 119 (17) ◽  
pp. 4066-4072 ◽  
Author(s):  
Bethan Psaila ◽  
James B. Bussel ◽  
Matthew D. Linden ◽  
Bracken Babula ◽  
Youfu Li ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Platelet Function ◽  
Whole Blood ◽  
Immune Thrombocytopenia ◽  
Ex Vivo ◽  
Platelet Reactivity ◽  
Adenosine Diphosphate ◽  
High Dose ◽  
Platelet Surface

Abstract The effects of eltrombopag, a thrombopoietin-receptor agonist, on platelet function in immune thrombocytopenia (ITP) are not fully characterized. This study used whole blood flow cytometry to examine platelet function in 20 patients receiving eltrombopag treatment at days 0, 7, and 28. Platelet surface expression of activated GPIIb/IIIa, P-selectin, and GPIb was measured with and without low and high adenosine diphosphate (ADP) and thrombin receptor activating peptide (TRAP) concentrations. Before eltrombopag treatment with no ex vivo agonist, platelet activation was higher in ITP patients than controls. Platelet GPIb and activated GPIIb/IIIa expression without added agonist was unchanged following eltrombopag treatment, whereas a slight increase in P-selectin was observed. Expression of P-selectin and activated GPIIb/IIIa in response to high-dose ADP was lower during eltrombopag treatment than at baseline. Eltrombopag led to a slight increase in platelet reactivity to TRAP only in responders to eltrombopag but not to levels above those in controls; whole blood experiments demonstrated that this increase was probably because of higher platelet counts rather than higher platelet reactivity. In conclusion, although thrombocytopenic ITP patients have higher baseline platelet activation than controls, eltrombopag did not cause platelet activation or hyper-reactivity, irrespective of whether the platelet count increased.

The control of blood platelets by cAMP signalling

2014 ◽  
Vol 42 (2) ◽  
pp. 289-294 ◽  
Author(s):  
Zaher Raslan ◽  
Khalid M. Naseem
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Platelet Activation ◽  
Blood Platelets ◽  
Blood Platelet ◽  
Platelet Inhibition ◽  
Signalling Pathway ◽  
Current Understanding ◽  
Camp Signalling ◽  
Kinase A

Blood platelet activation must be tightly regulated to ensure a balance between haemostasis and thrombosis. The cAMP signalling pathway is the most powerful endogenous regulator of blood platelet activation. PKA (protein kinase A), the foremost effector of cAMP signalling in platelets, phosphorylates a number of proteins that are thought to modulate multiple aspects of platelet activation. In the present mini-review, we outline our current understanding of cAMP-mediated platelet inhibition and discuss some of the issues that require clarification.

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.

Effect of Oestrogen Dose on Whole Blood Platelet Activation in Women Taking New Low Dose Oral Contraceptives

1994 ◽  
Vol 72 (06) ◽  
pp. 926-930 ◽  
Author(s):  
Lucy A Norris ◽  
John Bonnar
Keyword(s):  
Arachidonic Acid ◽  
Oral Contraceptive ◽  
Platelet Activation ◽  
Oral Contraceptives ◽  
Whole Blood ◽  
Contraceptive Use ◽  
Blood Platelet ◽  
Individual Treatment ◽  
Oral Contraceptive Use ◽  
Induced Aggregation

SummaryOral contraceptive use is known to cause changes in the haemostatic system. These changes are thought to be related to oestrogen dose and to provide a possible link between the increased risk of thromboembolic disease known to occur in women taking oestrogen containing oral contraceptives. This study measured whole blood platelet activation, serially, in women taking oral contraceptives containing 20 μg and 30 μg ethinyloestradiol combined with desogestrel. Increased levels of ADP and arachidonic acid induced aggregation were observed in women taking the 30 μg ethinyloestradiol combination. Platelet release of β-thromboglobulin (βTG) was also significantly increased. Increased collagen induced aggregation was observed but this failed to reach statistical significance for the individual treatment groups. In women taking the 20 μg ethinyloestradiol combination, a significant increase was only observed when platelets were stimulated with arachidonic acid. Platelet factor 4 (PF4) levels were unchanged in both groups. Significantly higher levels of βTG were observed in women taking the 30 μg ethinyloestradiol combination compared with women taking the 20 μg ethinyloestradiol combination. These results show that oral contraceptive use is associated with platelet activation. Women taking the 20 μg ethinyloestradiol combination show less changes in platelet activation than women taking the 30 μg ethinyloestradiol combination. This lower dose pill may therefore be particularly suitable for high risk women wishing to use oral contraception.

The Analysis of Mutual Relationship Between Hemostasis and Inflammatory Processes in Patients with Chronic Liver Diseases

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 1015-1015
Author(s):  
Nora E. El-Bassiouni ◽  
Rania M. Benyamin ◽  
Samia H. Risk ◽  
Manal Y. Zahran ◽  
Ola M. Mahmoud ◽  
...  
Keyword(s):  
Liver Cirrhosis ◽  
Platelet Count ◽  
Platelet Activation ◽  
Healthy Subjects ◽  
Blood Platelet ◽  
Surface Expression ◽  
Strongly Correlated ◽  
Blood Monocytes ◽  
Monocyte Activation ◽  
Platelet Surface

Abstract Introduction: Monocytes play a crucial role in the pathogenesis of inflammation and fibrosis in chronic liver diseases (CLD). Platelet, by connecting hemostasis and inflammatory processes, also participate in the pathogenesis of CLD. We aimed to assess the expression of platelet monocyte aggregates and their relationship to both monocyte and platelet activation in patients with hepatitis C virus (HCV) induced CLD in relation to the degree of hepatic insufficiency and haemostatic imbalance. Methods: Sixty patients with HCV induced CLD were categorized after Child Pugh criteria into 4 groups: Child A, B, C and C during acute attack of haematemesis, 15 patients each, 15 healthy subjects were also included as normal controls. Immunophenotype characterization for identification of platelet monocyte aggregates (CD41/CD45) on activated blood monocytes (CD11b and CD14) and activated blood platelet populations (CD61and CD62P) was carried out by flowcytometric analysis. The circulating levels of monocyte activation marker (MCP-1) and platelet activation markers (PF4 and sP-selectin) were also determined using immunological assays. Results: A marked reduction in platelet count concomitant with a decrease in the CD61 platelets surface expression were noticed in different groups of patients with HCV (p <0.01) compared to healthy subjects, implicating that the platelet count drops in correlation with advancement of liver cirrhosis. Data demonstrated a significant increase (p <0.01) in the surface expression of each of CD11b, CD14 and CD41/45 on peripheral blood monocytes revealing increased monocyte activation in patients with HCV compared to controls, especially Child C groups. These findings revealed that the increased expression of CD11b on blood monocytes matches the advancement of liver failure and that the CD14 is up regulated in viral hepatitis and correlate with liver injury. MCP-1 concentration did not correlate with monocyte count in peripheral blood denoting that MCP-1 level did not reflect the inflammatory activity in patients with liver cirrhosis, as it was lower in blood serum than in healthy subjects. Moreover, a significant progressive increase (p <0.01) in the expression of CD62P and CD41/CD45 on platelets and in the circulating levels of sP-selectin and PF4 was also detected in different groups of patients with HCV compared to controls. These findings revealed increased platelet activation with the rise in population of CD62P phenotype in patients with liver cirrhosis despite thrombocytopenia intensification. Our results also showed that the CD62P platelet surface expression was inversely correlated with platelet count and was strongly correlated with sP-selectin levels in Child B and C groups. Likewise, PF4 levels were strongly correlated with CD62P platelet surface expression and inversely correlated with platelet count in both groups revealing that the more developed thrombocytopenia, the stronger the platelet activation in liver cirrhosis. Our data also demonstrate that CD41/45 surface expression of blood platelet monocyte aggregate on platelets and monocytes are inversely correlated with platelet count and strongly correlated with each of monocyte activation (CD11b and CD14) and platelet activation (sP-selectin, CD62P and PF4) markers in patients with Child B and C showing that as cirrhosis develops and platelet count diminishes platelet aggregations with monocytes increase. Conclusions: Monocytes and platelets activation in patients with HCV increased in stepwise fashion in conjunction with worsening severity of liver disease as assessed by Child Pugh staging. The interplay of several factors such as endotoxaemia, lipopolysaccharides, oxidative stress and proinflammatory cytokines may be implicated in this respect. Also, the permanent flow of leukocytes and platelets through inflamed liver tissues and their long exposure to the active proinflammatory factors may lead to the activation of these cells. In turn, the progressive increase in monocytes and platelets activation with the simultaneous formation of monocyte-platelet aggregates noticed in patients with HCV may contribute to progressive liver injury in these patients. We, therefore, suggest that the blockage of platelet and monocyte activation may diminish thrombocytopenia and liver cirrhosis intensification in patients with chronic hepatitis C virus. Disclosures No relevant conflicts of interest to declare.

scholarly journals Comparative proteomics reveals unexpected quantitative phosphorylation differences linked to platelet activation state

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
G. J. Schmidt ◽  
C. M. Reumiller ◽  
H. Ercan ◽  
U. Resch ◽  
E. Butt ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Platelet Activation ◽  
Platelet Reactivity ◽  
Fold Increase ◽  
Surface Expression ◽  
Discrimination Power ◽  
Vasp Phosphorylation ◽  
Platelet Proteome ◽  
Quantitative Changes

AbstractThere is a need to assess platelet activation in patients with thrombotic disorders. P-selectin and activated integrin αIIbβ3 are usually quantified by flow cytometry to measure platelet activation. Monitoring changes in vasodilator-stimulated phosphoprotein (VASP) phosphorylation is an established method to determine the platelet-reactivity status. To study disruptions of platelet reactivity more comprehensively, we compared the human non-secretory platelet proteome after in-vitro -activation and –inhibition with their respective untreated controls using unbiased fluorescence two-dimensional differential in-gel electrophoresis. The non-secretory platelet proteome was more severely affected during inhibition than during activation. Strikingly, while VASP reached a 1.3-fold increase in phosphorylation levels in inhibited platelets, other protein kinase A targets showed several-fold stronger inhibition-induced phosphorylation levels, including LIM and SH3 domain protein 1 (6.7-fold), Src kinase-associated phosphoprotein 2 (4.6-fold), and Ras-related protein Rap1b (4.1-fold). Moreover, phosphorylation of integrin-linked protein kinase (ILK) and pleckstrin (PLEK) species was associated with P-selectin surface expression. The discrimination power between activation and inhibition was more pronounced for dephosphorylated ILK (3.79 Cohen’s d effect size) and phosphorylated PLEK (3.77) species than for P-selectin (2.35). These data reveal new insights into the quantitative changes of the platelet reactivity proteome and suggest powerful alternatives to characterise their activation and inactivation potential.

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