Abstract
Purpose: :
Proteases and protease inhibitors interplay are crucial events during wound healing. Here we study the modifications in LEI (Leukocyte Elastase Inhibitor) expression during wound healing in bovine corneal endothelial cells (BCEC), its dependence on the modality of healing (actin cable or lamellipoidial crawling) and on some well-recognized healing response initiators, such as the calcium and the reactive oxygen species (ROS) waves.
Methods: :
Wounds in BCEC monolayers were performed with a syringe needle, to remove, or a silicon-coated wire, to maintain the underlying ECM. Previously we have shown that BCEC heals by actin cable in the absence of ECM and by laemllipodial crawling in its presence. LEI expression was determined by IIF and Western Blot. The calcium wave (Fluo-4 staining) was inhibited by cyclopiazonic acid (CPA)/EDTA and the ROS wave by N-acetylcysteine (NAC).
Results: :
After incisional wounding in BCEC, there is a progressive rise in LEI at the wound border, that propagates several rows of cells towards the center of the monolayer, reaches its maximum at approximately 4 h and decreases as soon as the two margins re-establish contact. The LEI increase is larger under conditions of ECM preservation. The inhibition of the calcium or the ROS waves does not modify the LEI increase.
Conclusions: :
The increase in LEI expression suggests a role for this enzyme in the healing response. Moreover, the larger increase in lamellar zones may indicate a participation of the protein in cytoskeletal dynamics.
Keywords: cornea: endothelium • wound healing