Akt and mitogen-activated protein kinase enhance C-type lectin-like receptor2-mediated platelet activation by inhibition of glycogen synthase kinase 3 alpha/beta

作者：Moroi A J^*; Watson S P

来源：Journal of Thrombosis and Haemostasis, 2015, 13(6): 1139-1150.

DOI：10.1111/jth.12954

• C-type lectin
• glycogen synthase kinase 3
• mitogen-activated protein kinases
• phosphatidylinositol 3-kinases
• protein kinase B

BackgroundThe C-type lectin-like receptor2 (CLEC-2) and the collagen receptor glycoprotein (GP)VI activate platelets through Src and Syk tyrosine kinases, and phospholipaseC2. The initial events in the two signaling cascades, however, are distinct, and there are quantitative differences in the roles of proteins downstream of Syk activation. The activation of Akt and mitogen-activated protein kinases (MAPKs) has been shown to enhance platelet activation by GPVI, but their role in CLEC-2 signaling is not known.
ObjectivesWe sought to investigate the role of the Akt and MAPK pathways in platelet activation by CLEC-2.
ResultsThe CLEC-2 agonist rhodocytin stimulated phosphorylation of Akt and p38 and extracellular signal-related kinase (ERK) MAPKs, but with a delay relative to Syk. Phosphorylation of these proteins was markedly inhibited in the combined presence of apyrase and indomethacin, consistent with the reported feedback action of ADP and thromboxaneA(2) in CLEC-2 signaling. Phosphorylation of Akt and phosphorylation of ERK were blocked by the phosphoinositide 3-kinase (PI3K) inhibitor wortmannin and the protein kinaseC (PKC) inhibitor Ro31-8220, respectively, whereas Syk phosphorylation was not altered. On the other hand, both inhibitors reduced phosphorylation of the Akt substrate glycogen synthase kinase3/ (GSK3/). Phosphorylation of GSK3/ was also blocked by the Akt inhibitor MK2206, and reduced at late, but not early, times by the MEK inhibitor PD0325901. MK2206 and PD0325901 inhibited aggregation and secretion in response to a low concentration of rhodocytin, which was restored by GSK3/ inhibitors.
ConclusionsThese results demonstrate that CLEC-2 regulates Akt and MAPK downstream of PI3K and PKC, leading to phosphorylation and inhibition of GSK3/, and enhanced platelet aggregation and secretion.

• 出版日期2015-6