Dexamethasone inhibition of trabecular meshwork cell phagocytosis and its modulation by glucocorticoid receptor β

Xinyu Zhang, Cherie M. Ognibene, Abbot F. Clark, Thomas Yorio

Research output: Contribution to journalArticle

95 Scopus citations

Abstract

Glucocorticoid treatment can lead to the development of glaucomatous ocular hypertension and a secondary open-angle glaucoma due to increased aqueous humor outflow resistance that is associated with morphological and biochemical changes in the trabecular meshwork (TM). The cellular responses of glucocorticoids are achieved by binding to the glucocorticoid receptor α (GRα), a ligand-activated transcription factor. An alternatively spliced variant, glucocorticoid receptor β (GRβ), has dominant negative activity on GRα and has been implicated in a variety of steroid-resistant diseases. We previously showed that GRβ can block dexamethasone (DEX) responsiveness in TM cells. TM cells are actively phagocytic and function in the removal of debris, pigment and other materials from the aqueous outflow drainage pathway. A decrease in phagocytic activity has been proposed in the pathogenesis of glaucoma and glucocorticoid-induced glaucoma. In this study, we investigated the effect of DEX and GRβ on phagocytosis in normal and glaucomatous TM cells. Human transformed normal NTM-5 and primary normal NTM174-00 cells, which express relatively high amounts of GRβ, and transformed glaucomatous GTM-3 and primary glaucomatous GTM520-05 cells, which have lower GRβ expression, were treated with 100 nM DEX or vehicle control for 24 h. NTM cells also were transfected with a control or GRβ expression plasmid to examine the effect of GRβ on phagocytic activity. The cells were incubated with Alexa 488 conjugated Straphylococcus aureus bioparticles opsonized with rabbit IgG for 1 h, followed by fixation and incubation with Alexa 633 conjugated goat anti-rabbit IgG to distinguish ingested from extracellular bioparticles. DAPI nuclear staining was used to quantify cell numbers. Cells and bioparticles were visualized by confocal microscopy. We found that NTM-5 cells ingested more bioparticles than GTM-3 cells. DEX treatment significantly decreased the phagocytosis of bioparticles in NTM-5 and GTM-3 cells, while GTM-3 cells were more responsive to DEX, compared to NTM-5 cells. In primary cell culture, NTM174-00 also engulfed more bioparticles than GTM520-05 cells. DEX treatment significantly decreased the phagocytic activity in GTM520-05, but not in NTM174-00 cells. Transient transfection of pCMX-hGRβ plasmid increased the expression of GRβ and consequently maintained the phagocytotic activity of NTM-5 cells in the presence of DEX. Our data demonstrated that the expression level of GRβ in TM cells can regulate DEX-induced suppression of phagocytotic activity. The lower expression of GRβ in glaucomatous TM cells may contribute to the altered phagocytic function of TM cells, and may lead to the increased aqueous humor outflow resistance mediated by glucocorticoids.

Original languageEnglish
Pages (from-to)275-284
Number of pages10
JournalExperimental eye research
Volume84
Issue number2
DOIs
StatePublished - Feb 2007

Keywords

  • dexamethasone
  • glaucoma
  • glucocorticoid receptor β
  • phagocytosis
  • trabecular meshwork

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