›› 2013, Vol. 28 ›› Issue (5): 420-424.DOI: 10.3969/j.issn.1673-8640.2013.05.019

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Research on molecular mechanisms of PMA-induced platelet GPⅠbα shedding

  

  1. 1. Laboratory Center,Shanghai Traditional Chinese Medicine Hospital, Shanghai University of Traditional Chinese Medicine,Shanghai 200071, China; 2. Blood Engineering Laboratory,Shanghai Blood Center,Shanghai 200051, China
  • Received:2012-03-23 Revised:2012-04-17 Online:2013-05-15 Published:2013-05-14

Abstract: Objective Phorbol 12-myristate-13-acetate (PMA) could induce platelet GPⅠbα shedding,however,the molecular mechanisms are not fully elucidated. This study investigates the molecular mechanisms of PMA-induced platelet GPⅠbα shedding.  Methods Washed platelets were obtained from anti-coagulated healthy volunteers′ whole blood by centrifugation and washing. Washed platelets were pre-incubated with protein kinase C (PKC) inhibitor,reactive oxygen species (ROS) antagonist,NAD(P)H oxidase inhibitor,mitochondrial ROS antagonist,or dimethyl sulfoxide (DMSO),and were incubated with PMA. ROS levels were measured by flow cytometry. The production of platelet GPⅠbα shedding was detected by Western blot. The nonradioactive detection was used to measure PKC activity. Results PMA-induced platelet GPⅠbα shedding was completely inhibited by PKC inhibitor BIM or ROS antagonist dithiothreitol(DTT), respectively. PMA did dependently elevate ROS levels. PMA-induced ROS production was inhibited by PKC inhibitor and NAD(P)H oxidase inhibitor,but not mitochondrial ROS antagonist. BIM inhibited PMA-induced PKC activity,however,DTT did not. Conclusions PMA-induced platelet GPⅠbα shedding could be regulated by PMA-PKC-NAD(P)H oxidase-ROS-ADAM17-GPⅠbα signaling pathway.

Key words: Platelet, GPⅠb&alpha, shedding, Phorbol 12-myristate-13-acetate, Protein kinase C, Reactive oxygen species