Afsun Sahin1, Yang Liu2, Wendy R Kam2, Raheleh Rahimi Darabad3, David A Sullivan4. 1. Schepens Eye Research Institute of Massachusetts Eye and Ear and Department of Ophthalmology, Harvard Medical School, Boston, MA, USA; Department of Ophthalmology, Koc University Medical School, Istanbul, Turkey. 2. Schepens Eye Research Institute of Massachusetts Eye and Ear and Department of Ophthalmology, Harvard Medical School, Boston, MA, USA. 3. Schepens Eye Research Institute of Massachusetts Eye and Ear and Department of Ophthalmology, Harvard Medical School, Boston, MA, USA; Department of Clinical Anesthesia, Indiana University School of Medicine, Indianapolis, IN, USA. 4. Schepens Eye Research Institute of Massachusetts Eye and Ear and Department of Ophthalmology, Harvard Medical School, Boston, MA, USA. Electronic address: david_sullivan@meei.harvard.edu.
Abstract
PURPOSE: We discovered that dihydrotestosterone (DHT) decreases the ability of lipopolysaccharide, a bacterial toxin, to stimulate the secretion of leukotriene B4, a potent proinflammatory mediator, by immortalized human meibomian gland epithelial cells (IHMGECs). We hypothesize that this hormone action reflects an androgen suppression of proinflammatory gene activity in these cells. Our goal was to test this hypothesis. For comparison, we also examined whether DHT treatment elicits the same effect in immortalized human corneal (IHC) and conjunctival (IHConj) ECs. METHODS: Differentiated cells were cultured in media containing vehicle or 10 nM DHT. Cells (n = 3 wells/treatment group) were then processed for RNA isolation and the analysis of gene expression by using Illumina BeadChips, background subtraction, cubic spline normalization and Geospiza software. RESULTS: Our results demonstrate that DHT significantly suppressed the expression of numerous immune-related genes in HMGECs, such as those associated with antigen processing and presentation, innate and adaptive immune responses, chemotaxis, and cytokine production. DHT also enhanced the expression of genes for defensin β1, IL-1 receptor antagonist, and the anti-inflammatory serine peptidase inhibitor, Kazal type 5. In contrast, DHT had no effect on proinflammatory gene expression in HCECs, and significantly increased 33 gene ontologies linked to the immune system in HConjECs. CONCLUSIONS: Our findings support our hypothesis that androgens suppress proinflammatory gene expression in IHMGECs. This hormone effect may contribute to the typical absence of inflammation within the human meibomian gland.
PURPOSE: We discovered that dihydrotestosterone (DHT) decreases the ability of lipopolysaccharide, a bacterial toxin, to stimulate the secretion of leukotriene B4, a potent proinflammatory mediator, by immortalized human meibomian gland epithelial cells (IHMGECs). We hypothesize that this hormone action reflects an androgen suppression of proinflammatory gene activity in these cells. Our goal was to test this hypothesis. For comparison, we also examined whether DHT treatment elicits the same effect in immortalized humancorneal (IHC) and conjunctival (IHConj) ECs. METHODS: Differentiated cells were cultured in media containing vehicle or 10 nM DHT. Cells (n = 3 wells/treatment group) were then processed for RNA isolation and the analysis of gene expression by using Illumina BeadChips, background subtraction, cubic spline normalization and Geospiza software. RESULTS: Our results demonstrate that DHT significantly suppressed the expression of numerous immune-related genes in HMGECs, such as those associated with antigen processing and presentation, innate and adaptive immune responses, chemotaxis, and cytokine production. DHT also enhanced the expression of genes for defensin β1, IL-1 receptor antagonist, and the anti-inflammatory serine peptidase inhibitor, Kazal type 5. In contrast, DHT had no effect on proinflammatory gene expression in HCECs, and significantly increased 33 gene ontologies linked to the immune system in HConjECs. CONCLUSIONS: Our findings support our hypothesis that androgens suppress proinflammatory gene expression in IHMGECs. This hormone effect may contribute to the typical absence of inflammation within the human meibomian gland.
Authors: David A Sullivan; Eduardo M Rocha; Pasquale Aragona; Janine A Clayton; Juan Ding; Blanka Golebiowski; Ulrike Hampel; Alison M McDermott; Debra A Schaumberg; Sruthi Srinivasan; Piera Versura; Mark D P Willcox Journal: Ocul Surf Date: 2017-07-20 Impact factor: 5.033
Authors: E M Rocha; L A Wickham; L A da Silveira; K L Krenzer; F S Yu; I Toda; B D Sullivan; D A Sullivan Journal: Br J Ophthalmol Date: 2000-01 Impact factor: 4.638
Authors: Antje Schröder; Daniel B Abrar; Ulrike Hampel; Martin Schicht; Friedrich Paulsen; Fabian Garreis Journal: Exp Eye Res Date: 2016-08-26 Impact factor: 3.467
Authors: Di Chen; Afsun Sahin; Wendy R Kam; Yang Liu; Raheleh Rahimi Darabad; David A Sullivan Journal: Ocul Surf Date: 2018-05-31 Impact factor: 5.033
Authors: David A Sullivan; Benjamin D Sullivan; James E Evans; Frank Schirra; Hiroko Yamagami; Meng Liu; Stephen M Richards; Tomo Suzuki; Debra A Schaumberg; Rose M Sullivan; M Reza Dana Journal: Ann N Y Acad Sci Date: 2002-06 Impact factor: 5.691