PI3K signaling and lysyl oxidase is critical to corneal stroma fibrosis following mustard gas injury.

Abstract

Sulfur mustard gas (SM), an alkylating and vesicating agent, has been used frequently in many wars and conflicts. SM exposure to the eye results in several corneal abnormalities including scar/fibrosis formation. However, molecular mechanism for SM induced corneal fibrosis development is poorly understood. After SM insult to the eye, excessive synthesis/secretion of extracellular matrix components (ECM) including collagen (COL) I, COL III, and lysyl oxidase (LOX) by corneal myofibroblasts causes corneal fibrosis, however, precise mechanism remains elusive. This study tested the hypothesis that Phosphoinositide 3-kinase (PI3K) signaling alterations post SM in cornea enhances stromal ECM synthesis and corneal fibrosis. New Zealand White Rabbits were used. The right eyes were exposed to SM (200 mg-min/m³) and left eye to the air for 8min at MRI Global. Rabbit corneas were collected on day-3, day-7, and day-14 for molecular analysis. SM exposure caused a significant increase in mRNA expression of PI3K, AKT, COL I, COL III, and LOX and protein levels of LOX in a time-dependent manner in rabbit corneas. The in vitro studies were performed with human corneal stromal fibroblasts (hCSFs) by growing cultures in -/+ nitrogen mustard (NM) and LY294002, a PI3K specific inhibitor, for 30min, 8h, 24h, 48h, and 72h. NM significantly increased mRNA and protein levels of PI3K, AKT, COL I, COL III, and LOX. On the contrary, LY294002 in NM hCSFs significantly reduced PI3K, AKT, COL I, COL III, and LOX protein expression. We concluded that PI3K signaling mediates stromal collagen synthesis and LOX production following SM injury.