Platelets apoptosis in rats after global brain ischemia

Abstract

The purpose. Stroke is one of the most common causes of disability and mortality in the world. A variety of experimental models for the investigation of stroke focused on monitoring of platelet apoptosis. However, study and detection of platelet apoptosis in ischemia stroke in rats are very scarce. Using flow cytometry, we investigated the platelet apoptosis in rats after global brain ischemia. Methods. Experiments were carried out on healthy adult Wistar male rats (300–350 g). For modeling of global brain ischemia we used two-vessel (2-VO) model occlusion combined with hypotension. Platelets were isolated by gel filtration on Sepharose 2B. To detection of phosphatidylserine (PS) externalization platelets were incubated with Annexin V-PE and analyzed on the FACSCalibur (BD Biosciences). We used JC-1, a mitochondrial membrane potential indicator, to detect of mitochondrial membrane potential (∆ψ) during platelets apoptosis. After incubations platelets were analyzed by flow cytometry immediately. Results. Our results indicate that PS externalization on platelets after global brain ischemia in rats was significantly higher (53.45 ± 4.21%) as compared to control group of rats (5.27 ± 2.40%). This finding was confirmed by the expressed depolarization of the mitochondrial membrane potential (ΔΨm). Our results demonstrated that in platelets of rats after experimental brain ischemia, almost 40% (35.24 ± 5.21%) of platelets were depolarized as compared to control group of rats. Conclusion. Our results provide an insight into mechanisms that are involved in the platelet apoptosis during ischemic stroke. Finally, this study lays the foundation for future of new therapeutic strategies.