Abstract
Purpose :
To investigate the role of AMP-activated protein kinase (AMPK) in corneal epithelial cells stimulated by fungi.
Methods :
The non-toxic concentration range of AMPK phosphorylation agonist AICAR and inhibitor Compound C on corneal epithelial cells was screened by real-time marker-free multifunctional analyzer. Western Blot method was used to detect the expression of p-AMPK in corneal epithelial cells induced by AICAR and Compound C, and to screen the optimal concentration and time of drug action. The epithelial cells were divided into the corneal epithelial cell (C) group, corneal epithelial cell + spore (C+S) group, corneal ep-ithelial cell + spore + AICAR (C+S+A) group, corneal epithelial cell + spore + Compou-nd C ( C+S+CC) group. Western Blot method was used to detect the expression of AMPK and p-AMPK in corneal epithelial cells. ELISA method was used to detect the expression of IL-6 in the culture supernatant of corneal epithelial cells.
Results :
The cell index of 100μM, 300μM, 500μM, 1000 μM AICAR group and 10μM, 12.5μM Compound C group were not increased statistically than that in the corneal epithelial cells group at 24h, 36h, 48h, 72h time poingt (P >0.05) . The AICAR 1000μM group at 4h time point had the highest AMPK phosphorylation level in all AICAR concentration and time point groups (P<0.05). The AMPK phosphorylation level in the Compound C 10μM group was higher than that in the other Compound C concentration groups (P<0.05). Therefore , 1000μM and 4h was selected as the optimal concentration and time point for AICAR and 10μM group for Compound C. The AMPK phosphorylation level was higher in C+S group than that in group C (P<0.01). The AMPK phosphorylation levels were higher in the C+S+A group than that in the C+S group (P<0.05). The AMPK phosphorylation levels were lower in the C+S + CC group compared than that in the C+S group (P<0.05). The IL-6 expression in group C+S was higher in C+S group than that in group C ( P < 0.01) and C+S + CC group ( P < 0.05) .
Conclusions :
Corneal epithelial cells had AMPK phosphorylation expression, and the AMPK phosphorylation and IL-6 secretion were increased after fungal spores stimulate. AMPK phosphorylation and IL-6 may play an important role in corneal antifungal infection.
This is a 2021 ARVO Annual Meeting abstract.