Dihydrotestosterone Downregulates the Inflammatory Response in Human Conjunctival Epithelial Cells

Abstract

Abstract Purpose: The aim of our study is to investigate the effect of dihydrotestosterone (DHT) on the inflammatory response of the human conjunctival epithelial cells (HConjECs) under stress conditions. Methods: HConjECs were cultured with or without 10−8 M dihydrotestosterone (DHT) in normal osmolar conditions for 48 hours. Then, the media was changed to a higher osmolarity for different time points to demonstrate the effect of hyperosmolar stress conditions on both treated and untreated cells. After the cell pellet was lysed with radioimmunoprecipitation assay (RIPA), phosphorylated and nonphosphorylated forms of p38 kinase, extracellular signal-regulated kinase 1/2 (ERK1/2), and c-Jun N-terminal kinase 1/2 (JNK1/2) were analysed by western blot. The gene expressions of TNF-α, IL-1β, and IL-8 were measured with a real-time polymerase chain reaction. The cell viability was checked with Annexin V/PI. Results: Pretreatment with 10−8M DHT inhibited the hyperosmolarity-induced expression of TNF-α and IL-1β. Expression of p-P38 was inhibited with DHT pretreatment but only DHT-pretreated HConjECs suppressed p-ERK up to 30 minutes after hyperosmolar stress. DHT pretreatment inhibited apoptosis in HConjECs. Conclusion: These data revealed the potential importance of DHT in the treatment of DED. Furthermore, our findings may contribute to a better understanding of how sex hormones affect different ocular surface diseases and their potential for therapy.