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Caspase-3 gene expression in adenosine diphosphate stimulated platelets

Keywords: Platelets , caspase-3 , qRT-PCR , P-selectin , ADP

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Abstract:

Objectives: Platelets are known blood cells differentiatedfrom megakaryocytes in bone marrow and they playa key role in coagulation mechanism. They live severaldays in blood circulation and they aggregate in respondto stimulants such as thrombin, collagen, ADP (adenosinediphosphate) and epinephrine. Although platelets areknown anucleated cells, it has been reported in vivo andin vitro that they undergo to apoptosis. In this study apoptoticeffects of ADP on the platelets was investigated.Materials and methods: P-selectin binding with flowcytometry,calcium levels with fluorometer, cytochrome-clevels with ELISA, caspase-3 gene expression with qRTPCR,caspase-3 releasing with western blotting were determinedin platelets.Results: Platelets were incubated for 1 hour at 37°C with10μM ADP. P-selectin binding, calcium and cytochromeclevels, caspase-3 releasing and gene expression aresignificantly increased compared to control in ADP stimulatedplatelets.Conclusions: Platelets can induce caspase-3 gene expressionwhen stimulated with ADP which is a key enzymeplays an important role in apoptotic pathway. J ClinExp Invest 2012; 3 (3): 372-377Key words: Platelets, caspase-3, qRT-PCR, P-selectin,ADP

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