ABSTRACT
Purpose: The purpose of this study is to explore the effects of dihydrotestosterone (DHT) on lipopolysaccharide (LPS)-induced proinflammatory cytokine release in human ocular surface epithelial cells exposed to LPS and LPS-binding protein (LBP).
Methods: Immortalized human corneal, conjunctival, and meibomian gland epithelial cells were cultured in keratinocyte-free medium. After confluency, they were exposed to a stratification medium Dulbecco's modified Eagle medium (DMEM)/F12 in the presence of fetal bovine serum and were exposed to vehicle, LPS + LBP, or DHT. Culture media were processed for multiplex-bead analysis of specific proinflammatory cytokines including interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-2, IL-4, IL-8, IL-6, IL-10, IL-1β, vascular endothelial growth factor (VEGF)-A. Cytokine concentrations were compared by analysis of variance with Tukey post hoc testing. p < 0.05 was considered statistically significant.
Results: The results are LPS + LBP-induced the secretion of IFN-γ, IL-6, IL-10, IL-1β, VEGF-A cytokines in corneal epithelial cells; TNF-α, IL-2, IL-8, IL-6, IL-1β, VEGF-A cytokines in conjunctival epithelial cells; and IL-8, IL-6, IL-1β, VEGF-A cytokines in meibomian gland epithelial cells. When these LPS + LBP-stimulated cells were exposed to DHT for 2 days, it was found that DHT suppressed the secretion of IL-6, IL-10, IL-1β, VEGF-A cytokines in corneal epithelial cells; TNF-α, IL-6, IL-1β, VEGF-A cytokines in conjunctival epithelial cells; and IL-6, IL-1β, VEGF-A cytokines in meibomian gland epithelial cells.
Conclusion: LPS + LBP is shown to induce the secretion of certain proinflammatory cytokines from ocular surface and adnexal epithelial cells. DHT showed anti-inflammatory activity by suppressing some of those cytokines in these cell lines.
Acknowledgments
Additional contributions: Ilene K. Gipson, PhD, Schepens Eye Research Institute, Harvard Medical School, provided the immortalized human conjunctival cells; James Jester, PhD, University of California, Irvine, provided the corneal epithelial cells; and David Sullivan, PhD Schepens Eye Research Institute, Harvard Medical School, provided the meibomian gland epithelial cells.
Disclosure statement
The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the article.