Abstract
Purpose :
NF-κB is a key transcription factor responsible for many biological processes, such as inflammation, apoptosis, stress response, corneal wound healing, angiogenesis, and lymphangiogenesis. In the present study, we tried to explore the effect of BAY-11, a NF-kB inhibitor, on human conjunctival epithelium cells (HCjE) in response to LPS stimulation in culture.
Methods :
Human conjunctival epithelial cells (HCjEs) were cultured in DMEM medium with 10% FBS for 24 hours containing 25 ng/ml LPS, with or without 0.5 µM BAY-11. Cell viability was assessed using Olympus live imaging microscope and DAPI staining. Intracellular ROS was measured in HCjEs using the fluorescent probe H2-DCF. Surface expression of ICAM-1 in HCjEs exposed to LPS (25ng/ml) or LPS with NF-kB inhibitor BAY-11 (0.5 µM) was evaluated after 24 hours incubation using a cell modified ELISA .
Results :
BAY-11 had no measurable cytotoxic effect on HCjEs wheras stimulation of the cells with LPS showed a significant reduction of the cell number. When BAY-11 was added this effect was partially abolished and a higher cell survival rate – compared to the cells that were treated with LPS alone – was observed. There was no significant increase in ROS production or ICAM-1 surface expression in the cell samples cultured with BAY-11alone. In contrast LPS stimulation of the cells led to a significant increase (p<0.05) of ROS production, accompanied by a correspondingly high upregulation of ICAM-1 surface expression (p<0.05). However, in the presence of BAY-11 the ROS production and ICAM-1 levels were significantly reduced.
Conclusions :
In conclusion, BAY-11 significantly improved viability and reduced ICAM-1 surface expression and ROS production after LPS stimulation in HCjEs. These findings indicate that BAY-11 could have therapeutic potential against oxidative stress-related and inflammatory disorders of the ocular surface.
This is a 2020 ARVO Annual Meeting abstract.