Role of Calcium in Platelet Activation: Novel Insights and Pharmacological Implications

Davlouros, Periklis; Xanthopoulou, Ioanna; Mparampoutis, Nikolaos; Γιαννόπουλος, Γεώργιος; Deftereos, Spyridon; Alexopoulos, Dimitrios

doi:10.2174/157340641202160208195923

Cited by 30 publications

(22 citation statements)

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“…Calcium mobilization is a very dynamic process, susceptible to rapid change in response to stimulus and therefore a good marker to detect changes in platelet signalling. It governs not only platelet activation, but also secretion and aggregation [ 38 ]. It is therefore understandable that subtler changes are possible to register with appropriately sensitive measurement of calcium flux and its inhibition.…”

Section: Discussionmentioning

confidence: 99%

Adenosine Receptor Agonists Increase the Inhibition of Platelet Function by P2Y12 Antagonists in a cAMP- and Calcium-Dependent Manner

et al. 2020

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We have shown previously that platelet activity can be lowered through the simultaneous inhibition of P2Y12 receptor and activation of adenosine receptors (AR). This work explores this concept by testing the antiplatelet potential of nine AR agonists in combination with P2Y12 receptor antagonists—cangrelor and prasugrel metabolite. A panel of in vitro methods was used to assess platelet viability, P-selectin expression, GPIIb-IIIa activation, fibrinogen binding, calcium ion mobilization, VASP-P level, and cAMP formation, utilizing whole blood or isolated platelets from healthy volunteers. The AR agonists demonstrated anti-platelet effects, but stimulated signaling pathways to varying degrees. AR agonists and P2Y12 antagonists reduced expression of both P-selectin and the activated form of GPIIb-IIIa on platelets; however, the combined systems (AR agonist + P2Y12 antagonist) demonstrated stronger effects. The antiplatelet effects of AR when combined with P2Y12 were more pronounced with regard to exogenous fibrinogen binding and calcium mobilization. The cAMP levels in both resting and ADPactivated platelets were increased by AR agonist treatment, and more so when combined with P2Y12 inhibitor. In conclusion, as AR agonists are fast-acting compounds, the methods detecting early activation events are more suitable for assessing their antiplatelet action. The exogenous fibrinogen binding, calcium mobilisation and cAMP level turned out to be sensitive markers for detecting the inhibition caused by AR agonists alone or in combination with P2Y12 receptor antagonists.

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Section: Discussionmentioning

confidence: 99%

Adenosine Receptor Agonists Increase the Inhibition of Platelet Function by P2Y12 Antagonists in a cAMP- and Calcium-Dependent Manner

et al. 2020

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“…In our hypothesis, EDTA has been repeatedly claimed to be inappropriate for PRP preparation, as EDTA might functionally disrupt platelets by chelating Ca 2+ involved in many functions [32], and their growth factors may be lost or reduced from PRP prepared from EDTA-anticoagulated whole-blood samples. As predicted, platelet functions were suppressed to some extent; however, their growth factors seemed to be more preserved than predicted.…”

Section: Discussionmentioning

confidence: 89%

“…The Fluo4 + platelet percentage was lower in EDTA-and heparin-anticoagulated samples than that in ACD-A-anticoagulated samples; however, the difference was not significant. Figure 6 shows the effects of anticoagulants on intra-platelet free Ca 2+ concentrations, which is thought to influence platelet activity and functions [32]. The representative histograms of Fluo4 + platelets reveal that the overall fluorescence intensity in EDTA-anticoagulated samples was somewhat lower than that in the ACD-A-anticoagulated samples.…”

Section: Discussionmentioning

confidence: 99%

A Comparative Study of the Effects of Anticoagulants on Pure Platelet-Rich Plasma Quality and Potency

et al. 2020

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It is generally accepted that citrate or the A-form of acid-citrate-dextrose (ACD-A) are suitable for preparing platelet-rich plasma (PRP) for regenerative therapy. However, this is based on evidence from blood transfusions and not from regenerative medicine. Thus, we examined the effects of anticoagulants, such as ACD-A, ethylenediaminetetraacetic acid (EDTA), and heparin, on the regenerative quality of PRP to address this gap. The blood samples were collected in the presence of anticoagulants and were processed to prepare pure-PRP. Platelet size, activation status, and intra-platelet free Ca2+ concentration were determined while using a hematology analyzer and flow cytometer. Platelet-derived growth factor-BB (PDGF-BB) was quantified while using an ELISA. In pure-PRP samples, EDTA caused platelet swelling and activation, but yielded the highest number of platelets. Heparin aggregated platelets and disturbed the overall counting of blood cells. However, no significant differences in PDGF-BB levels were observed among the anticoagulants tested. Moreover, when considering the easy preparation of platelet suspensions, without the need for high-level pipetting skills, these findings suggest the comparable potency of EDTA-derived pure-PRP in tissue regeneration and support the use of EDTA in the preparation of pure-PRP. Further in vivo studies are required in animal models to exclude the possible negative effects of including EDTA in pure-PRP preparations.

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“…In our dataset, we observe that several of altered proteins, and associated pathways, were associated with functions that may be modulated by calcium. For example, platelet activation is strongly dependent on an increase of intracellular Ca 2+ concentration [42] . Similarly, in K562 cells, exosome release is regulated by a calcium-dependent mechanism [43] .…”

Section: Resultsmentioning

confidence: 99%

Multiplexed Isobaric Tag‐Based Profiling of Seven Murine Tissues Following In Vivo Nicotine Treatment Using a Minimalistic Proteomics Strategy

et al. 2018

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Nicotine is a major addictive compound in tobacco and a component of smoking-related products, such as e-cigarettes. Once internalized, nicotine can perturb many cellular pathways and can induce alterations in proteins across different cell types, however the mechanisms thereof remain undetermined. We hypothesize that both tissue-specific and global protein abundance alterations result from nicotine exposure. As such, we present the first proteome analysis of multiple tissues from nicotine-exposed mice. We treat mice via oral administration of nicotine at 200μg/mL in drinking water for 21 days. We investigate 7 tissues (brain, heart, kidney, liver, lung, pancreas, and spleen) from treated (n=5) and untreated control (n=5) mice. For each tissue, a TMT10-plex experiment (5 versus 5) was assembled. We apply a minimalistic proteomics strategy was employed using TMT reagents efficiently and fractionating by centrifugation-based reversed-phase columns to streamline sample preparation. Combined, we quantified over 11,000 non-redundant proteins from over 138,000 different peptides in 7 TMT10-plex experiments. Between 7 and 126 proteins are significantly altered in tissues from nicotine-exposed mice. Among these proteins, only 11 are altered in two or more tissues, many classified as from extracellular exosomes or involved in lipid metabolism. Our data showcase the vast extent of nicotine exposure across murine tissue.

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Role of Calcium in Platelet Activation: Novel Insights and Pharmacological Implications

Cited by 30 publications

References 0 publications

Adenosine Receptor Agonists Increase the Inhibition of Platelet Function by P2Y12 Antagonists in a cAMP- and Calcium-Dependent Manner

Adenosine Receptor Agonists Increase the Inhibition of Platelet Function by P2Y12 Antagonists in a cAMP- and Calcium-Dependent Manner

A Comparative Study of the Effects of Anticoagulants on Pure Platelet-Rich Plasma Quality and Potency

Multiplexed Isobaric Tag‐Based Profiling of Seven Murine Tissues Following In Vivo Nicotine Treatment Using a Minimalistic Proteomics Strategy

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