LOX-1 and TLR4 affect each other and regulate the generation of ROS in A. fumigatus keratitis

Purpose To explore the relationship between LOX-1 and TLR4 in Aspergillus fumigatus (A. fumigatus) keratitis. To determine LOX-1 and TLR4 can affect each other and regulate inflammation through regulation of the generation of reactive oxygen species (ROS) in A. fumigatus keratitis. Methods The cornea and abdominal cavity extracted neutrophils of susceptible C57BL/6 mice were infected with A. fumigatus. The cornea and neutrophils were pretreated with LOX-1 neutralizing antibody, Polyinosinic acid (Poly(I)) (the inhibitor of LOX-1) or CLI-095 (the inhibitor of TLR4) separately before infection. LOX-1, TLR4 and IL-1β expression were detected in normal and infected cornea by PCR and Western Blot, while ROS was detected in the neutrophils by flow cytometry. Results LOX-1, TLR4, IL-1β mRNA and protein levels were up-regulated in C57BL/6 cornea after infection. LOX-1 neutralizing antibody or Poly(I) pretreatment decreased the expression of LOX-1, TLR4 and IL-1β in C57BL/6 cornea after infection and CLI-095 pretreatment decreased the expression of LOX-1, TLR4 and IL-1β in C57BL/6 cornea after infection. ROS generation was increased in C57BL/6 neutrophils after infection, however, ROS generation was decreased in C57BL/6 neutrophils after infection by LOX-1 neutralizing antibody or Poly(I) or CLI-095 pretreatment. Conclusion LOX-1, TLR4 and IL-1β expression and ROS generation are increased after infection. LOX-1 and TLR4 can affect each other and regulate the generation of ROS in A. fumigatus keratitis. Inhibition of LOX-1 and TLR4 can reduce ROS generation.