Intraplatelet reactive oxygen species (ROS) correlate with the shedding of adhesive receptors, microvesiculation and platelet adhesion to collagen during storage: Does endogenous ROS generation downregulate platelet adhesive function?

Ghasemzadeh, Mehran; Hosseini, Ehteramolsadat; Roudsari, Zahra Oushyani; Zadkhak, Parvin

doi:10.1016/j.thromres.2018.01.048

Cited by 32 publications

(35 citation statements)

References 64 publications

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“…A). We have already found increasing levels of PS exposure during storage, with an observed significant increase from day 5, and this observation was confirmed in this study (Fig. B).…”

Section: Resultssupporting

confidence: 91%

“…Studies showed that during platelet storage, microvesiculation can be caused by the ignition of the apoptotic pathways, especially in long‐stored platelets, whereas the blockade of platelet activation during storage had only a partial effect on MP generation . Several lines of evidence have suggested a role for ROS in the induction of MP formation, whereas our previous study showed a direct relation between ROS generation and MP formation during storage . Data presented here demonstrated a significant reduction in platelet microvesiculation in 5‐day‐stored platelets in the presence of both NAC and VAS2870.…”

Section: Discussioncontrasting

confidence: 58%

“…36 Several lines of evidence have suggested a role for ROS in the induction of MP formation, whereas our previous study showed a direct relation between ROS generation and MP formation during storage. 20 Data presented here demonstrated a significant reduction in platelet microvesiculation in 5-day-stored platelets in the presence of both NAC and VAS2870. However, in 7-day-stored platelets, VAS2870 treatment did not significantly reduce MP formation, whereas NAC protected platelets from microvesiculation even more significantly.…”

Section: Discussionsupporting

confidence: 49%

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ROS scavenger, N‐acetyl‐l‐cysteine and NOX specific inhibitor, VAS2870 reduce platelets apoptosis while enhancing their viability during storage

et al. 2019

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BACKGROUND Platelet storage is often complicated by deleterious changes that are started by reversible activation of the cells and can lead to procoagulant function and apoptosis during longer periods of storage. Given that increasing levels of reactive oxygen species (ROS) generation are associated with platelet activation and apoptosis, our study investigated whether ROS scavenging or the inhibition of ROS production can enhance the viability of stored platelets. METHODS For this study platelet‐rich plasma platelet concentrates (PRP‐PCs) were either treated with ROS‐reducing agents (1 mM N‐acetyl‐L‐cysteine [NAC] or 30 μM NADPH oxidase [NOX] inhibitor, VAS2870) or kept untreated during storage. P‐selectin expression, phosphatidylserine (PS) exposure, levels of microparticle (MP) formation, and intraplatelet caspase activity of PCs were analyzed by flow cytometry during 7 days of storage while the platelet viability was also evaluated by MTT assay. RESULTS Both NAC‐ and VAS2870‐treated platelets had significantly lower caspase activity, MP formation, and PS exposure during storage while they also showed improved viability. The platelet count and mean platelet volume (MPV) were also better preserved in the presence of NAC. CONCLUSION Our results confirmed that either the inhibition of ROS generation or the scavenging of these oxidant agents can attenuate platelet apoptosis while improving their viability during storage. In this study, the significant improvement of platelet viability obtained by NAC suggested that its supplementation with a designated safe concentration into PCs may better preserve the quality of these products, especially for longer storage.

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“…A). We have already found increasing levels of PS exposure during storage, with an observed significant increase from day 5, and this observation was confirmed in this study (Fig. B).…”

Section: Resultssupporting

confidence: 91%

Section: Discussioncontrasting

confidence: 58%

Section: Discussionsupporting

confidence: 49%

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ROS scavenger, N‐acetyl‐l‐cysteine and NOX specific inhibitor, VAS2870 reduce platelets apoptosis while enhancing their viability during storage

et al. 2019

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“…Oxidative stress is the dysregulated generation of reactive oxygen species (ROS) and has been shown to cause platelet hyperactivity leading to thrombosis. [1][2][3][4] NADPH oxidases (NOXs) are important sources of ROS in platelets [5][6][7] and their function promotes platelet activation. 8,9 NOXs are a family of multimeric plasma membrane complexes generating the majority of non-mitochondrial ROS in mammalian cells.…”

Section: Introductionmentioning

confidence: 99%

NADPH oxidase 1 is a novel pharmacological target for the development of an antiplatelet drug without bleeding side effects

et al. 2020

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Growing evidence supports a central role of NADPH oxidases (NOXs) in the regulation of platelets, which are circulating cells involved in both hemostasis and thrombosis. Here, the use of Nox1−/− and Nox1+/+ mice as experimental models of human responses demonstrated a critical role of NOX1 in collagen‐dependent platelet activation and pathological arterial thrombosis, as tested in vivo by carotid occlusion assays. In contrast, NOX1 does not affect platelet responses to thrombin and normal hemostasis, as assayed in tail bleeding experiments. Therefore, as NOX1 inhibitors are likely to have antiplatelet effects without associated bleeding risks, the NOX1‐selective inhibitor 2‐acetylphenothiazine (2APT) and a series of its derivatives generated to increase inhibitory potency and drug bioavailability were tested. Among the 2APT derivatives, 1‐(10H‐phenothiazin‐2‐yl)vinyl tert‐butyl carbonate (2APT‐D6) was selected for its high potency. Both 2APT and 2APT‐D6 inhibited collagen‐dependent platelet aggregation, adhesion, thrombus formation, superoxide anion generation, and surface activation marker expression, while responses to thrombin or adhesion to fibrinogen were not affected. In vivo administration of 2APT or 2APT‐D6 led to the inhibition of mouse platelet aggregation, oxygen radical output, and thrombus formation, and carotid occlusion, while tail hemostasis was unaffected. Differently to in vitro experiments, 2APT‐D6 and 2APT displayed similar potency in vivo. In summary, NOX1 inhibition with 2APT or its derivative 2APT‐D6 is a viable strategy to control collagen‐induced platelet activation and reduce thrombosis without deleterious effects on hemostasis. These compounds should, therefore, be considered for the development of novel antiplatelet drugs to fight cardiovascular diseases in humans.

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“…Generally, deleterious changes, so-called as platelet storage lesion (PSL) affects the quality and effectiveness of therapeutic platelets. The storage of PCs is associated with progressive platelet activation characterized by accumulating intra-platelet ROS, adhesive receptors shedding, platelet procoagulant activity and granule release leading to potential pro-in ammatory function of platelets [34], [35,36]. Pro-coagulant activity (monitored by PS exposure) is also associated with apoptotic events, including cytochrome C release, caspase 3 activation, and the loss of mitochondrial respiration capacity as speci c markers of apoptosis, however the signi cant rises of these markers are usually detected in long-stored platelets, from day 5 or 7 of storage [25].…”

Section: Discussionmentioning

confidence: 99%

Collagen-dependent platelet dysfunction and its relevance to either mitochondrial ROS or cytosolic superoxide generation: a question about the quality and functional competence of long-stored platelets

Hosseini

Hojjati

gashti

et al. 2020

Preprint

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Background: Upon vascular damage, the exposed subendothelial matrix recruits circulating platelets to site of injury while inducing their firm adhesion mainly via GPVI-collagen interaction. GPVI also supports aggregatory and pro-coagulant functions in arterial shear rate even on the matrix other than collagen. Reactive oxygen species (ROS) modulate these stages of thrombosis; however augmented oxidant stress also disturbs platelet functions. Stored-dependent platelet lesion is associated with the increasing levels of ROS. Whether ROS accumulation is also relevant to collagen-dependent platelet dysfunction is the main interest of this study. Methods: Fresh PRP-PCs (platelet concentrates) were either stimulated with potent ROS-inducers PMA and CCCP or stored for 5 days. Intra-platelet superoxide (O2--) or mitochondrial-ROS and GPVI expression were detected by flowcytometery. GPVI shedding, platelet aggregation and spreading/adhesion to collagen were analyzed by western blot, aggregometry and fluorescence-microscopy respectively. Results: Mitochondrial-ROS levels in 5 days-stored PCs were comparable to those induced by mitochondrial uncoupler, CCCP while O2-- generations were higher than those achieved by PMA. Shedding levels in 5 days-stored PCs were higher than those induced by these potent stimuli. GPVI expressions were reduced comparably in CCCP treated and 5 days-stored PCs. Platelet adhesion was also diminished during storage while demonstrating significant reverse correlation with GPVI shedding. However, only firm adhesion (indicated by spreading or platelet adhesion surface area) was relevant to GPVI expression. Platelet adhesion and aggregation also showed reverse correlations with both O2-- and mitochondrial-ROS formations; nonetheless mitochondrial-ROS was only relevant to firm adhesion. Conclusion: As a sensitive indicator of platelet activation, GPVI shedding correlated with either simple adhesion or spreading to collagen, while GPVI expression was only relevant to platelet spreading. Thereby, if the aim of GPVI evaluation is to examine platelet firm adhesion, expression seems to be a more specific choice. Furthermore, the comparable levels of ROS generation in 5 days-stored PCs and CCCP treated platelets, indicated that these products are significantly affected by oxidative stress. Reverse correlation of accumulating ROS with collagen-dependent platelet dysfunction is also a striking sign of an oxidant-induced lesion that may raise serious question about the post-transfusion quality and competence of longer stored products.

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Intraplatelet reactive oxygen species (ROS) correlate with the shedding of adhesive receptors, microvesiculation and platelet adhesion to collagen during storage: Does endogenous ROS generation downregulate platelet adhesive function?

Cited by 32 publications

References 64 publications

ROS scavenger, N‐acetyl‐l‐cysteine and NOX specific inhibitor, VAS2870 reduce platelets apoptosis while enhancing their viability during storage

ROS scavenger, N‐acetyl‐l‐cysteine and NOX specific inhibitor, VAS2870 reduce platelets apoptosis while enhancing their viability during storage

NADPH oxidase 1 is a novel pharmacological target for the development of an antiplatelet drug without bleeding side effects

Collagen-dependent platelet dysfunction and its relevance to either mitochondrial ROS or cytosolic superoxide generation: a question about the quality and functional competence of long-stored platelets

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