title: Genomic and post-genomic effects of anti-glaucoma drugs preservatives in trabecular meshwork.
abstract: Oxidative stress plays an important role in glaucoma. Some preservatives of anti-glaucoma drugs, commonly used in glaucoma therapy, can prevent or induce oxidative stress in the trabecular meshwork. The aim of this study is to evaluate cellular and molecular damage induced in trabecular meshwork by preservatives contained in anti-glaucoma drugs. Cell viability (MTT test), DNA fragmentation (Comet test), oxidative DNA damage (8-oxo-dG), and gene expression (cDNA microarray) have been evaluated in trabecular meshwork specimens and in human trabecular meshwork cells treated with benzalkonium chloride, polyQuad, purite, and sofzia-like mixture. Moreover, antimicrobial effectiveness and safety of preservative contents in drugs was tested. In ex vivo experiments, benzalkonium chloride and polyQuad induced high level of DNA damage in trabecular meshwork specimens, while the effect of purite and sofzia were more attenuated. The level of DNA fragmentation induced by benzalkonium chloride was 2.4-fold higher in subjects older than 50 years than in younger subjects. Benzalkonium chloride, and polyQuad significantly increased oxidative DNA damage as compared to sham-treated specimens. Gene expression was altered by benzalkonium chloride, polyQuad, and purite but not by sofzia. In in vitro experiments, benzalkonium chloride and polyQuad dramatically decreased trabecular meshwork cell viability, increased DNA fragmentation, and altered gene expression. A lesser effect was also exerted by purite and sofzia. Genes targeted by these alterations included Fas and effector caspase-3. The efficacy of the preservatives in inhibiting bacterial growth increased the adverse effects in trabecular meshwork in terms of DNA damage and alteration of gene expression. Presented data indicates the delicate balance between efficacy and safety of drug preservatives as not yet optimized.
authors.affiliation: Mutagenesis Unit, IRCCS AOU San Martino - IST, Genova, Italy; Department of Health Sciences, University of Genoa, Via A. Pastore 1, 16132 Genoa, Italy. Electronic address: izzotti@unige.it.
authors.forename: Alberto
authors.initials: A
authors.lastname: Izzotti
authors.affiliation: Department of Health Sciences, University of Genoa, Via A. Pastore 1, 16132 Genoa, Italy.
authors.forename: Sebastiano
authors.initials: S
authors.lastname: La Maestra
authors.affiliation: Department of Health Sciences, University of Genoa, Via A. Pastore 1, 16132 Genoa, Italy.
authors.forename: Rosanna Tindara
authors.initials: RT
authors.lastname: Micale
authors.affiliation: Department of Health Sciences, University of Genoa, Via A. Pastore 1, 16132 Genoa, Italy.
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Longobardi
authors.affiliation: Ophthalmology Unit, IRCCS AOU San Martino-IST, Genova, Italy.
authors.forename: Sergio Claudio
authors.initials: SC
authors.lastname: Saccà
mesh_terms: D000328:Adult; D000368:Aged; D000891:Anti-Infective Agents, Local / Q000494:pharmacology*; D001548:Benzalkonium Compounds / Q000494:pharmacology*; D002460:Cell Line; D004249:DNA Damage / Q000187:drug effects*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D005901:Glaucoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D011108:Polymers / Q000494:pharmacology*; D014129:Trabecular Meshwork / Q000378:metabolism* / Q000473:pathology
chemical_list: D000891:Anti-Infective Agents, Local; D001548:Benzalkonium Compounds; D011108:Polymers; C058655:polyquaternium 1
keywords: Drug preservatives; Gene expression; Glaucoma; Microarray; Oxidative damage; Trabecular meshwork