Phospholipase C/Protein Kinase C Pathway is Essential for Corneal Re-epithelialization Induced by Ap4A

Purpose: We have previously demonstrated the importance of P2Y2 receptors in the corneal re-epithelialization effect triggered by diadenosine tetraphosphate (Ap4A). In addition, we have also shown the ERK1/2 and ROCK-I activation in Ap4A-wound repair response. Phospholipase C/Protein Kinase C (PLC/PKC) pathway activation has been suggested as a molecular mechanism of growth factors-modulated corneal cell migration and P2Y2 agonists. Hence, the aim of this study is to investigate the role of PLC/PKC cascade in the modification of re-epithelialization rate triggered by Ap4A in an established corneal epithelial cell line (Statens Seruminstitut rabbit cornea [SIRC] cells).Methods: In wounded confluent SIRC cell monolayers and in the presence or absence of Ap4A 100 µM, a group of PLC/PKC inhibitors (U73122 3 µM, Staurosporine 1 nM and Bisindolylmaleimide-I 10 µM) and activator (PDBU 1 µM) were assayed and the migration rate was evaluated. Also, the activation of ERK1/2 and ROCK-I was examined by Western blot a...