alexa Thrombin induces apoptotic events through the generation of reactive oxygen species in human platelets.
Medicine

Medicine

Anatomy & Physiology: Current Research

Author(s): Lopez JJ, Salido GM, GmezArteta E, Rosado JA, Pariente JA

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Abstract BACKGROUND: Thrombin is a major physiological platelet agonist that activates a number of cell functions including aggregation. Platelet stimulation with thrombin has been shown to result in the development of apoptotic events, including activation of caspases-3 and -9, cytochrome c release and phosphatidylserine (PS) exposure; however, the mechanism underlying the activation of apoptosis remains unclear. OBJECTIVES: In the present study, we aim to investigate whether endogenously generated reactive oxygen species upon thrombin stimulation is required for the activation of apoptosis in human platelets. METHODS: Changes in the mitochondrial membrane potential were registered using the dye JC-1; caspase-3 and -9 activity was determined from the cleavage of their respective specific fluorogenic substrates; PS externalization was estimated using annexin V-fluorescein isothicyanate and cytochrome c release was detected by Western blotting in samples from the mitochondrial and cytosolic fractions. RESULTS: Treatment of platelets with thrombin stimulates mitochondrial membrane potential depolarization and endogenous generation of H(2)O(2) . Platelet exposure to exogenous H(2)O(2) results in cytochrome c release and activation of caspases-9. In addition, H(2)O(2) induces the activation of caspase-3 and PS exposure by a mechanism dependent on cytochrome c release and caspase-9 activation. Finally, thrombin-evoked development of apoptotic events was impaired by treatment with catalase. CONCLUSION: Our results indicate that thrombin-induced apoptosis is likely mediated by endogenous generation of H(2)O(2) in human platelets. This article was published in J Thromb Haemost and referenced in Anatomy & Physiology: Current Research

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