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.

scholarly journals Preparations from selected cucurbit vegetables as antiplatelet agents

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Agata Rolnik ◽  
Bartosz Skalski ◽  
Anna Stochmal ◽  
Beata Olas
Keyword(s):  
Platelet Activation ◽  
Cucurbita Pepo ◽  
Fruits And Vegetables ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
Antiplatelet Agents ◽  
Blood Platelet ◽  
Arachidonic Acid Metabolism ◽  
Platelet Activity ◽  
Activated Platelets

AbstractIncreased 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 (Cucurbita 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 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.

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.

Supplemental Fibrinogen Restores Platelet Inhibitor-Induced Reduction in Thrombus Formation without Altering Platelet Function: An In Vitro Study

2020 ◽  
Vol 120 (11) ◽  
pp. 1548-1556
Author(s):  
Thomas Bärnthaler ◽  
Elisabeth Mahla ◽  
Gabor G. Toth ◽  
Rufina Schuligoi ◽  
Florian Prüller ◽  
...  
Keyword(s):  
Antiplatelet Therapy ◽  
Platelet Activation ◽  
Platelet Function ◽  
Dual Antiplatelet Therapy ◽  
Thrombus Formation ◽  
In Vitro Study ◽  
Coronary Intervention ◽  
Calcium Flux ◽  
Major Surgery

Abstract Background For patients treated with dual antiplatelet therapy, standardized drug-specific 3-to-7 day cessation is recommended prior to major surgery to reach sufficient platelet function recovery. Here we investigated the hypothesis that supplemental fibrinogen might mitigate the inhibitory effects of antiplatelet therapy. Methods and Results To this end blood from healthy donors was treated in vitro with platelet inhibitors, and in vitro thrombus formation and platelet activation were assessed. Ticagrelor, acetylsalicylic acid, the combination of both, and tirofiban all markedly attenuated the formation of adherent thrombi, when whole blood was perfused through collagen-coated microchannels at physiological shear rates. Addition of fibrinogen restored in vitro thrombus formation in the presence of antiplatelet drugs and heparin. However, platelet activation, as investigated in assays of P-selectin expression and calcium flux, was not altered by fibrinogen supplementation. Most importantly, fibrinogen was able to restore in vitro thrombogenesis in patients on maintenance dual antiplatelet therapy after percutaneous coronary intervention. Conclusion Thus, our in vitro data support the notion that supplementation of fibrinogen influences the perioperative hemostasis in patients undergoing surgery during antiplatelet therapy by promoting thrombogenesis without significantly interfering with platelet activation.

A Significant Fraction of ADAMTS13 Activity Is Associated with Activated Platelets and Their Microparticles (PMP): Implication for Regulating ADAMTS13 Activity.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1066-1066
Author(s):  
Wenche Jy ◽  
Andrew Lin ◽  
Loreta Bidot ◽  
Jaehoon Bang ◽  
Eugene Ahn ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Control Level ◽  
In Vitro Study ◽  
Ionophore A23187 ◽  
Adamts13 Activity ◽  
Activated Platelets ◽  
Membrane Bound ◽  
Low Levels

Abstract BACKGROUND: Deficiency of ADAMTS13, vWF cleaving protease, is known to be associated with TTP and some other microangiopathies, but low levels were reported in other diseases such as ITP, DIC, lupus and other thromboses. Although inhibitory autoantibodies were demonstrated in TTP, the mechanisms underlying reduced levels of ADAMTS13 in other disorders remains unclear. We tested the hypothesis that ADAMTS13 is associated with cell membranes and derived microparticles, especially from activated platelet and their microparticles (PMP), which could modulate the enzyme activities of ADAMTS13. METHODS: PRP was prepared by centrifuging citrated normal blood for 10 min at 160×g, and PPP by further centrifuging for 10 min at 3,000×g, and particle-free plasma (PFP) by further centrifuging for 15 min at 20,000×g. ADAMTS13 activity was assayed by the FRETS-VWF73 method of Kokame et al [Br J Haematol 129:93, 2005] using the Fluoroskan Ascent plate reader. Platelets were activated by ADP (10 μM) or ionophore A23187 (2 μM). RESULTS: The ADAMTS13 activity (A) of pooled PPP of 10 controls was defined as 100%. (1) In vitro study: (1a) ADAMTS13 activity was not significantly different between PPP and resting PRP. However, if the platelets in PRP were first activated by ADP for 1hr, a significant reduction of activity was observed (A = 85 ±7%, p<0.05). If the activated platelets were removed, the activity of the supernatant fell to 79 ±10% p<0.05) of the control level, and was further reduced by higher centrifugation to remove PMP (A = 66 ±12%, p<0.01). (1b) Activation by A23187, a stronger agonist producing 2–3 fold more PMP than ADP (confirmed by flow cytometry), induced a more dramatic reduction in PRP (A = 78±8%, p<0.01), and after removal of platelets (A = 71 ±11%, p<0.01), and after removal of PMP (48 ±11%, p<0.01). (1c) Interestingly, resuspending the activated platelets did not restore ADAMTS13 activity, although resuspending the PMP did partially restore the activity. (2) In vivo study: PPP from 13 patients (6 ITP, 4 APS, 3 lupus) were analyzed. The majority (11/13) of PPP samples lost activity after removal of PMP (A = 79 ±12% in PPP vs. 64 ±11% in PFP; p <0.02). CONCLUSION: These data show that a significant but variable fraction of ADAMTS13 activity is associated with activated platelets and PMP. This has several implications. First, distinguishing soluble from membrane-bound ADAMTS13 may lead to better correlation of activities with clinical findings, and may help explain low levels of ADAMTS13 in some disorders associated with platelet activation and high PMP. Second, this interaction may play a role in regulating ADAMTS13 activity. Third, membrane-bound ADAMTS13 may clear more readily from circulation, therefore inhibiting platelet activation or MP formation may have benefits for the management of microangiopathies.

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.

scholarly journals TREM-like transcript 1: a more sensitive marker of platelet activation than P-selectin in humans and mice

2018 ◽  
Vol 2 (16) ◽  
pp. 2072-2078 ◽  
Author(s):  
Christopher W. Smith ◽  
Zaher Raslan ◽  
Lola Parfitt ◽  
Abdullah O. Khan ◽  
Pushpa Patel ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Thrombus Formation ◽  
Surface Expression ◽  
Activated Platelets ◽  
Key Points ◽  
Sensitive Marker

Key Points Platelet activation in vitro results in a more rapid and greater upregulation of TLT-1 surface expression compared with P-selectin. TLT-1 is more rapidly translocated to the surface of activated platelets than P-selectin during thrombus formation in vivo.

The Role of ROCK in Platelet–Monocyte Collaborative Induction of Thromboinflammation during Acute Coronary Syndrome

2020 ◽  
Vol 120 (10) ◽  
pp. 1417-1431
Author(s):  
Ling-Wei Hsu ◽  
Po-Wei Chen ◽  
Wei-Ting Chang ◽  
Wen-Huang Lee ◽  
Ping-Yen Liu
Keyword(s):  
Acute Coronary Syndrome ◽  
Coronary Arteries ◽  
Cardiovascular Events ◽  
Blood Platelets ◽  
Coiled Coil ◽  
Ribosome Profiling ◽  
Platelet Activity ◽  
Coronary Syndrome ◽  
Activated Platelets

Abstract Background Arterial thrombosis is initiated by atherosclerotic plaque damage, prothrombotic material release and platelet aggregation. Platelets are primary mediators involved in thrombosis and cooperate with vascular and immune cells. Objective Herein, we investigated how activated platelets interacted with monocytes in atherothrombosis. Methods and Results We collected patients' blood from coronary arteries during percutaneous coronary intervention and measured platelet activity. Platelets from coronary arteries had higher pseudopodium expression and activity in patients with acute coronary syndrome (ACS). Ribosome profiling of platelets from coronary blood mapped a vigorous upregulation of Rho GTPases and their downstream effectors. RhoA activated downstream Rho-associated coiled-coil containing protein kinase (ROCK), and ROCK increased surface P-selectin in coronary blood platelets. The interaction between platelets and monocytes was observed in vitro, and was found in ruptured coronary plaques of ACS. Further we found that activated platelets promoted monocytes transmigration, which could be suppressed in the presence of ROCK inhibitors. The increased surface P-selectin on thrombin-induced platelets interacted with monocytes to upregulate monocyte chemokine receptor 2 (CCR2) expression via the ROCK pathway. The expression of CCR2 was higher in monocyte–platelet aggregates than in monocytes without platelets. Finally, using the Asian Screening Array BeadChip, we identified single-nucleotide polymorphism (SNP) associated with cardiovascular events. Notably, patients having homozygous major alleles of the RHOA SNP rs11706370 presented with higher risks of cardiovascular events. Conclusion Through ROCK-activated cytoskeleton remodeling and P-selectin expression, platelets were recruited and interacted synergistically with high CCR2-expressing monocytes to induce thromboinflammation in atherothrombosis.

The influence of different rewetting procedures on the thrombogenicity of nanoporous poly(ether imide) microparticles

2020 ◽  
pp. 1-14
Author(s):  
S. Braune ◽  
J. Bäckemo ◽  
S. Lau ◽  
M. Heuchel ◽  
K. Kratz ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
Saline Group ◽  
Vinyl Pyrrolidone ◽  
Covalent Attachment ◽  
Reference Ranges ◽  
Chemical Treatments ◽  
Phosphate Buffered Saline

Nanoporous microparticles prepared from poly(ether imide) (PEI) are discussed as candidate adsorber materials for the removal of uremic toxins during apheresis. Polymers exhibiting such porosity can induce the formation of micro-gas/air pockets when exposed to fluids. Such air presenting material surfaces are reported to induce platelet activation and thrombus formation. Physical or chemical treatments prior to implantation are discussed to reduce the formation of such gas nuclei. Here, we report about the influence of different rewetting procedures – as chemical treatments with solvents – on the thrombogenicity of hydrophobic PEI microparticles and PEI microparticles hydrophilized by covalent attachment of poly(vinyl pyrrolidone) (PVP) of two different chain lengths. Autoclaved dry PEI particles of all types with a diameter range of 200 – 250 μm and a porosity of about 84% ±2% were either rewetted directly with phosphate buffered saline (24 h) or after immersion in an ethanol-series. Thrombogenicity of the particles was studied in vitro using human sodium citrated whole blood (60 min, 5 rpm vertical rotation). Numbers of non-adherent platelets were quantified, and adhesion of blood cells was qualitatively analyzed by bright field microscopy. Platelet activation (percentage of CD62P positive platelets and amounts of soluble P-Selectin) and platelet function (PFA100 closure times) were analysed. Retention of blood platelets on the particles was similar for all particle types and both rewetting procedures. Non-adherent platelets were less activated after contact with ethanol-treated particles of all types compared to those rewetted with phosphate buffered saline as assessed by a reduced number of CD62P-positive platelets and reduced amounts of secreted P-Selectin (P < 0.05 each). Interestingly, the hydrophilic surfaces significantly increased the number of activated platelets compared to hydrophobic PEI regardless of the rewetting agent. This suggests that, apart from wettability, other material properties might be more important to regulate platelet activation. PFA100 closure times were reduced and within the reference ranges in the ethanol group, however, significantly increased in the saline group. No substantial difference was detected between the tested surface modifications. In summary, rewetting with ethanol resulted in a reduced thrombogenicity of all studied microparticles regardless of their wettability, most likely resulting from the evacuation of air from the nanoporous particles.

Different Effects of Ranibizumab and Bevacizumab on Platelet Activation Profile

2015 ◽  
Vol 234 (4) ◽  
pp. 195-210 ◽  
Author(s):  
Bianka Sobolewska ◽  
Cornelia Grimmel ◽  
Aikaterini Gatsiou ◽  
Kateryna Sopova ◽  
Judith Klein ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Platelet Activation ◽  
Light Transmission ◽  
In Vitro Study ◽  
Cellular Level ◽  
High Dose ◽  
Activation Markers ◽  
Activated Platelets ◽  
Increased Risk

Purpose: The aim of the study was to evaluate the potential influence of ranibizumab and bevacizumab on platelet activation and aggregation, which are critical processes in the pathogenesis of arterial thromboembolic events (ATEs). Methods: For the assessment of platelet function, flow cytometry and aggregometry were employed. Platelets were isolated from healthy volunteers and exposed to ranibizumab (1 mg/ml and 150 ng/ml) and bevacizumab (2.5 mg/ml and 3 μg/ml) or their solvents for 10 and 30 min prior to the addition of TRAP (25 μM), PAR-4-AP (25 μM) or thrombin (0.02 U/ml). The surface expression of activated GP IIb/IIIa, P-selectin (CD62P) and platelet-bound stromal cell-derived factor-1 (SDF-1) was measured on resting (nonactivated) and activated platelets by flow cytometry. The platelet aggregation capacity was examined using light transmission aggregometry. Results: The expression of surface activation markers did not differ significantly between nonstimulated and TRAP-, PAR-4-AP- or thrombin-activated platelets after incubating with ranibizumab. However, GP IIb/IIIa, CD62P and SDF-1 were significantly downregulated in PAR-4-AP- and thrombin-activated platelets after exposure to bevacizumab 2.5 mg/ml. In addition, ranibizumab- and bevacizumab-FITC were significantly increased in all activated platelets. No significant differences were observed in the aggregation of activated platelets after incubation with ranibizumab or bevacizumab. Conclusion: All ranibizumab concentrations as well as the bevacizumab concentration of 3 μg/ml had no influence on platelet activation and aggregation. Therefore, this in vitro study did not show any relationship between the exposition of activated platelets to ranibizumab or bevacizumab and the development of ATEs. However, the highest level of bevacizumab interfered with platelet activation, leading to downregulation of platelet activation markers. This observation might explain why the systemic treatment with high-dose bevacizumab could be associated with an increased risk of bleeding. Regarding the use of lower intravitreal dosages, further research should focus on the complex interactions between platelets and other cells, such as endothelial cells, which might stronger relate to a potentially increased risk of ATEs and depend on systemic vascular endothelial growth factor levels. Facing the different activation profiles, the diverse effects of the drugs on the cellular level have to be critically scrutinized for their clinical relevance.

scholarly journals Cangrelor Attenuates Coated-Platelet Formation

2008 ◽  
Vol 15 (2) ◽  
pp. 177-182 ◽  
Author(s):  
Nicholas B. Norgard ◽  
Callie L. Hann ◽  
George L. Dale
Keyword(s):  
Platelet Activation ◽  
In Vitro Study ◽  
Pharmacological Agents ◽  
Platelet Production ◽  
Activated Platelets ◽  
Prothrombinase Activity ◽  
Dual Agonist ◽  
Platelet Formation

P2Y12 inhibitors were introduced clinically as effective inhibitors of adenosine-5′-diphosphate (ADP) mediated platelet activation and aggregation. This class of pharmacological agents has enjoyed considerable success. Cangrelor is a recently developed P2Y12 inhibitor that has the advantage of being an active drug not requiring metabolic conversion, although it is not orally available. Coated-platelets are a subclass of activated platelets generated on dual agonist activation with collagen plus thrombin; the primary hallmark of coated-platelets is their ability to support prothrombinase activity. Interestingly, we recently observed that the relatively weak agonist ADP potentiates the production of coated-platelets by the very strong agonists collagen plus thrombin, a previously unknown role for ADP. The authors sought in this study to determine if P2Y12 inhibitors, such as cangrelor, were capable of attenuating this augmentation of coated-platelet generation. Cangrelor, at physiologically relevant concentrations, was able to eliminate the ADP-dependent increase in coated-platelet production with an IC50 of 1.4 nM. Cangrelor, however, had no effect on thrombin-dependent platelet activation as measured by P-selectin expression. Although this in vitro study does not address the question of whether the effectiveness of cangrelor in vivo is partially due to an attenuation of coated-platelet production in addition to its documented antiaggregatory effects, it does reveal an unexpected action of cangrelor. Additional studies will be required to determine if all P2Y12 inhibitors are equally effective in attenuating coated-platelet production.

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