- 作者：Armen V. Gyulkh ; anyan ; Asuman Mutlu ; David J. Allen ; John Freedman ; Valery Leytin
- 关键词：PRP ; platelet-rich plasma ; 螖唯m ; mitochondrial inner transmembrane potential ; MP ; microparticle ; PS ; phosphatidylserine ; DMSO ; dimethylsulfoxide ; GPRP ; Gly-Pro-Arg-Pro peptide ; DiOC6(3) ; 3 ; 3&rsquo ; -dihexyloxacarbocyanine iodide ; FSC ; forward scatter ; SSC ; side scatter
- 刊名：Thrombosis Research
- 出版年：January, 2014
- 年：2014
- 卷：133
- 期：1
- 页码：73-79
- 全文大小：796 K
Background
Depolarization of mitochondrial inner transmembrane potential (螖唯m) is a key biochemical manifestation of the intrinsic apoptosis pathway in anucleate platelets. Little is known, however, about the relationship between 螖唯m depolarization and downstream morphological manifestations of platelet apoptosis, cell shrinkage and microparticle (MP) formation.Objectives
To elucidate this relationship in human platelets.
Materials and Methods
Using flow cytometry, we analyzed 螖唯m depolarization, platelet shrinkage and MP formation in platelets treated with BH3-mimetic ABT-737 and calcium ionophore A23187, well-known inducers of intrinsic platelet apoptosis.
Results
We found that at optimal treatment conditions (90 min, 37 掳C) both ABT-737 and A23187 induce 螖唯m depolarization in the majority (88-94%) of platelets and strongly increase intracellular free calcium. In contrast, effects of A23187 and ABT-737 on platelet shrinkage and MP formation are quite different. A23187 strongly stimulates cell shrinkage and MP formation, whereas ABT-737 only weakly induces these events (10-20% of the effect seen with A23187, P < 0.0001).
Conclusions
These data indicate that a high level of 螖唯m depolarization and intracellular free calcium does not obligatorily ensure strong platelet shrinkage and MP formation. Since ABT-737 efficiently induces clearance of platelets from the circulation, our results suggest that platelet clearance may occur in the absence of the morphological manifestations of apoptosis.