Effects of Preserved and Preservative-Free Glaucoma Drugs on Proteomic Expression Levels in Corneal and Conjunctival Epithelial Cells in vitro
Year: 2015
Type: dissertation
Abstract: Benzalkonium chloride (BAC), is the most commonly used preservative world-wide in cosmetics and eye drops; however, it is toxic to epithelial cells and the pathways in this toxicity are largely unknown.In this study, the effects of common ophthalmic drugs for glaucoma were uncovered by exposing human corneal and conjunctival epithelial cells (HCE and IOBA-NHC respectively) to treatments with and without BAC.It was hypothesized that these drugs may be linked to inflammatory mechanisms and cell death.The aim of this study was to explore proteomic data in order to discover some potential biomarkers relating to BAC-induced effects, which could be beneficial in further studies.HCE and IOBA-NHC cells were exposed to a preservative-free topical medication tafluprost, a similar drug latanoprost which contains BAC or BAC by itself for 24 hours and then the proteomic profiles of treated and untreated cells were analysed with NanoLC-TOF-MS using SWATH technique.Central tendency normalization was applied to the log2-transformed proteomic data once the quality of the data was initially ensured with descriptive statistics including correlation and clustering methods.Mixed-effects ANOVA model was implemented to data to uncover differentially Abbreviation Meaning ACG Angle closure glaucoma ANOVA Analysis of variance ARACNE Algorithm for the Reconstruction of Accurate Cellular Networks BAC Benzalkonium chloride BP Biological process CAI Carbonic anhydrase inhibitors CC Cellular component DAVID Database for Annotation, Visualization and Integrated Discovery DIA Data-independent acquisition ES Enrichment score FDR False discovery rate GO Gene ontology GOrilla Gene Ontology enRIchment anaLysis and visuaLization tool GSEA Gene set enrichment analysis HCE Human corneal epithelial cells HK2 Hexokinase-2 HMGCS1 Hydroxymethylglutaryl-CoA synthase, cytoplasmic IOBA-NHC Immortalized normal human conjunctival epithelial cells IOP Intraocular pressure IPI International protein index iTRAQ Isobaric tag for relative and absolute quantitation KEGG Kyoto encyclopaedia of genes and genomes LC Liquid chromatography MF Molecular function MI Mutual information MLC Myosin light chain MRM Multiple reaction monitoring MS Mass spectrometry MYH9 Myosin-9 MYL12A Myosin regulatory light chain 12A MYL6 Myosin light polypeptide 6 NDUFA5 NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 5 NDUFS3 NADH dehydrogenase [ubiquinone] iron sulphur protein 3, mitochondrial OAG Open angle glaucoma OSD Ocular surface disease ROS Reactive oxygen species SRM Selected reaction monitoring SWATH Sequential window acquisition of all theoretical spectra WST-1 Water-soluble tetrazolium salt
Author: Nättinen, Janika
Language: English
Topic: Drug-Induced Ocular Toxicity
Subfield: Ophthalmology
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W883127403