Lipoxin A4 inhibits platelet-activating factor inflammatory response and stimulates corneal wound healing of injuries that compromise the stroma

Platelet-activating factor (PAF) is a bioactive lipid mediator with strong inflammatory properties. PAF induces the expression and activation of metalloproteinase-9 (MMP-9) in corneal epithelial cells and myofibroblasts, and delays epithelial wound healing in an organ culture system. Lipoxin A4 (LXA4) is a lipid mediator involved in resolution of inflammation and cornea epithelial wound healing. We developed an in vivo mouse model of injury to the anterior stroma that is sustained by PAF and evaluated the action of LXA4. In this model mice were treated with vehicle, PAF alone and in combination with PAF receptor antagonist LAU-0901 or LXA4. Mice were euthanized 1, 2 and 7 days after injury and corneas were processed for histology (H&E staining) and immunofluorescence with antibodies for MMP-9, α-smooth muscle actin (α-SMA), fibronectin (FN) and neutrophil. Interleukin 1-α (IL-1α) and keratinocyte-derived chemokine (KC/CXCL1) were assayed by ELISA. Myeloperoxidase (MPO) activity was performed in corneal homogenates. In this in vivo model PAF inhibited epithelial wound healing that was blocked by the PAF receptor antagonist LAU-0901. Treatment with LXA4 significantly reduced the injured area compared to PAF at 1 and 2 days of treatment. The strong stromal cell infiltration and MPO activity stimulated by PAF was also decreased with LXA4 treatment. PAF increased MMP-9 and decreased FN expression compared to vehicle treatment and less α-SMA positive cells migrated to the wounded area. The PAF actions were reverted by LXA4 treatment. The results demonstrated a powerful action of LXA4 in protecting corneas with injuries that compromise the stroma by decreasing inflammation and increasing wound healing. ► Platelet-activating factor is a lipid with strong inflammatory properties. ► Lipoxin A4 is a lipid that mediates resolution of inflammation. ► We investigate the interaction between these mediators in an in vivo mouse model. ► Treatment with LXA4 reduced the injured area and PAF-induced MMP-9 expression. ► LXA4 protects corneas with deep injures exacerbated by PAF.