title: In vitro effects of sex hormones in human meibomian gland epithelial cells.
abstract: Meibomian gland dysfunction (MGD) is considered the most common cause of dry eye disease (DED). Sex hormones seem to play a role in the pathogenesis of MGD although their involvement is not completely understood. Therefore, in the present study we evaluated the effect of dihydrotestosteron (DHT) and estradiol (β-Est) on an immortalized human meibomian gland epithelial cell line (HMGEC). Protein expression of sex hormone receptors in HMGEC was investigated by western blot. Ultrastructural morphology, Sudan III lipid staining, cell proliferation as well as vitality assays were performed. Furthermore, expression of MGD-associated markers for keratinization (hornerin, involucrin and CK6), proliferation (CK5 and CK14) and lipid synthesis (fatty acid synthase and stearoyl-CoA desaturase) were analyzed by real time RT-PCR. Western blot revealed presence of androgen receptor (AR), estrogen receptors α and -β (ERα, ERβ) and progesterone receptor (PR) in HMGEC. PR, ERα and ERβ expression was significantly induced under cultivation with serum, whereas sex hormones stimulation showed no further effect on protein expression of PR, ERα and ERβ. Our results showed no impact of MGD-associated sex hormones to cellular morphology and lipid accumulation in HMGEC. Cell proliferation was slightly induced through application of sex hormones and supplementation of calcium. However, both sex hormones and calcium altered gene expression of MGD-associated markers. Especially keratinization genes hornerin (HRNR) and cornulin (COR) were induced after application of sex hormones and calcium in serum-free cultivated HMGEC. This may promote keratinization processes that are associated with MGD. Further investigations are necessary to analyze the (hyper)keratinization processes that occur during MGD and using HMGEC as an in vitro model.
authors: Department of Anatomy II, Friedrich Alexander University Erlangen-Nürnberg (FAU), Erlangen, Germany.
authors: Antje
authors: A
authors: Schröder
authors: Department of Anatomy II, Friedrich Alexander University Erlangen-Nürnberg (FAU), Erlangen, Germany.
authors: Daniel B
authors: DB
authors: Abrar
authors: Department of Anatomy II, Friedrich Alexander University Erlangen-Nürnberg (FAU), Erlangen, Germany; Department of Ophthalmology, Gutenberg University Mainz, Germany.
authors: Ulrike
authors: U
authors: Hampel
authors: Department of Anatomy II, Friedrich Alexander University Erlangen-Nürnberg (FAU), Erlangen, Germany.
authors: Martin
authors: M
authors: Schicht
authors: Department of Anatomy II, Friedrich Alexander University Erlangen-Nürnberg (FAU), Erlangen, Germany.
authors: Friedrich
authors: F
authors: Paulsen
authors: Department of Anatomy II, Friedrich Alexander University Erlangen-Nürnberg (FAU), Erlangen, Germany. Electronic address: fabian.garreis@fau.de.
authors: Fabian
authors: F
authors: Garreis
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D015352:Dry Eye Syndromes / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D004847:Epithelial Cells / Q000187:drug effects* / Q000648:ultrastructure; D005786:Gene Expression Regulation; D012739:Gonadal Steroid Hormones / Q000494:pharmacology*; D006801:Humans; D008537:Meibomian Glands / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics; D011960:Receptors, Estrogen / Q000096:biosynthesis / Q000235:genetics; D011980:Receptors, Progesterone / Q000096:biosynthesis / Q000235:genetics
chemical_list: D012739:Gonadal Steroid Hormones; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D012313:RNA
keywords: Androgen; Dry eye disease (DED); Estradiol; Hornerin (HRNR); Human meibomian gland epithelial cell line (HMGEC); Meibomian gland dysfunction (MGD); Ocular surface; Tear film