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

Abstract

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.

Compound Prunetin Induces Cell Death in Gastric Cancer Cell with Potent Anti-Proliferative Properties: In Vitro Assay, Molecular Docking, Dynamics, and ADMET Studies

“…Necrosomes have been reported to impair mitochondrial energy metabolism by disturbing ROS homeostasis, leading to cell death [ 42 ]. A growing number of reports also suggest that ROSs could disturb the mitochondrial disturbance in the event of inducing necroptosis cell death [ 44 , 45 ]. To uncover the relation of ROS generation through JNK activation, the amount of intracellular singlet oxygen generated in AGS cells treated with PRU (0, 20, 40, and 80 μM) was measured with or without specific JNK inhibitor (SP100125) at 10 μM concentration.…”

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

“…The storage of PCs is associated with progressive platelet activation characterized by accumulating intra-platelet ROS, adhesive receptors shedding, platelet pro-coagulant activity and granule release leading to potential pro-inflammatory function of platelets [ 34 – 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 specific markers of apoptosis, however the significant rises of these markers are usually detected in long-stored platelets, from day 5 or 7 of storage [ 25 ]. Altogether, these are the stored-dependent changes of platelet which not only affect post-transfusion platelet survival but it may also attenuate platelet functional activity and effectiveness in circulation, while of note, some of these changes such as receptor shedding and the induction of pro-inflammatory or pro-coagulant phenotypes of platelets sound to be irreversible.…”

“…So far, several studies indicated that increasing levels of ROS generation in stored platelets is associated with PSL. Most recently, we have indicated that in stored platelets, either the ROS scavenging or the reduction of their generation by NOX inhibition, can effectively promote platelet viability while reducing PSL effect during storage [25]. Now, considering the role of oxidative stress in the modulation of GPVI, here we also investigated whether increasing levels of ROS during platelet storage can be functionally relevant to the storage-dependent loss of platelet spreading/ adhesion to collagen.…”

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

“…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]. Altogether, these are the stored-dependent changes of platelet which not only affect post-transfusion platelet survival but it may also attenuate platelet functional activity and effectiveness in circulation, while of note, some of these changes such as receptor shedding and the induction of pro-in ammatory or pro-coagulant phenotypes of platelets sound to be irreversible.…”