Abstract
Purpose :
In the present study, we aim to elucidate the underlying molecular mechanism to better understand endoplasmic reticulum (ER) stress induced delayed corneal epithelial wound healing and nerve regeneration.
Methods :
Human limbal epithelial cells (HLECs) were treated with thapsigargin to induce excessive ER stress and then RNA sequencing was performed. Immunofluorescence and qPCR were used to detect the expression changes of SLIT3 and its receptors Robo1-4 in mouse corneal epithelium and cultured corneal epithelial epithelial stem/progenitor cells. The role of recombinant SLIT3 protein in corneal epithelial proliferation and migration were assessed by CCK8 and cell scratch assay, respectively. Thapsigargin, exogenous SLIT3 protein and ROBO4 siRNA was injected subconjunctivally to evaluate the effects of different intervention on corneal epithelial and nerve regeneration.
Results :
Thapsigargin suppressed normal corneal epithelial and nerve regeneration significantly. RNA sequencing genes related to development and regeneration revealed that thapsigargin induced ER stress significantly upregulated the expression of SLIT3 and ROBO4 in corneal epithelial cells. Exogenous SLIT3 inhibited normal corneal epithelial injury repair and nerve regeneration, and significantly suppressed the proliferation and migration ability of cultured mouse corneal epithelial cells. ROBO4 siRNA significantly attenuated the delayed corneal epithelial injury repair and nerve regeneration induced by SLIT3 treatment and ERS.
Conclusions :
ER stress inhibits corneal epithelial injury repair and nerve regeneration maybe related with the activation of SLIT3-ROBO4 pathway.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.