title: Lens epithelial cell protection by aminothiol WR-1065 and anetholedithiolethione from ionizing radiation.
abstract: Lens epithelium disorganization, glutathione (GSH) depletion, and epithelial cell death have been incriminated in the cytopathogenic mechanisms that lead to cataract formation following UVB and x-ray exposures. The objective of this study was to determine the in vitro capacity of the aminothiol WR-1065, the active metabolite of amifostine, and anetholedithiolethione (ADT or Sulfarlem) to protect bovine lens epithelial cells against x-ray irradiation. WR-1065 and ADT were used at a concentration of 20 microM. A single dose of 10 Gy was delivered at a rate of 2 Gy/min. Fluorimetric assays were then performed using a neutral red probe to evaluate cell viability, a Hoechst 33342 probe (HO) to evaluate nuclear condensation and apoptosis, and a monobromobimane probe to estimate the intracellular GSH pool. Twenty-four hours after x-ray exposure, cells pretreated with WR-1065 showed increased GSH levels, improved cell viability, and decreased HO fluorescence in addition to a lesser proportion of cells with apoptotic nuclear modifications. Between 72 and 120 hr postirradiation, ADT-pretreated cells also showed increased intracellular GSH levels and cell viability and decreased HO fluorescence and apoptotic cell morphology. This in vitro study demonstrates that WR-1065 and ADT protects lens epithelial cells from x-ray injury; thus, ADT and amifostine are appropriate candidates for clinical trials in humans. They are currently used in preventing radiation-induced xerostomia and should be further tested in the prevention of late radiation-induced ocular complications such as sicca syndrome and cataract.
authors: Cell Pharmacotoxicology Unit, Department of Pharmacy, CHNO des XV-XX, and Laboratory of Toxicology, UFR Pharmacy, University of Paris V, France. yazid.belkacemi@tnn.ap-hop-paris.fr
authors: Y
authors: Y
authors: Belkacémi
authors: P
authors: P
authors: Rat
authors: G
authors: G
authors: Piel
authors: M O
authors: MO
authors: Christen
authors: E
authors: E
authors: Touboul
authors: J M
authors: JM
authors: Warnet
mesh_terms: D004999:Amifostine / Q000494:pharmacology; D000781:Anethole Trithione / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis; D001562:Benzimidazoles / Q000494:pharmacology; D002417:Cattle; D016923:Cell Death; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D004307:Dose-Response Relationship, Radiation; D004847:Epithelial Cells / Q000187:drug effects*; D005434:Flow Cytometry; D005978:Glutathione / Q000378:metabolism; D007908:Lens, Crystalline / Q000187:drug effects*; D008624:Mercaptoethylamines / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D011837:Radiation-Protective Agents / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014466:Ultraviolet Rays; D014965:X-Rays
chemical_list: D001562:Benzimidazoles; D008624:Mercaptoethylamines; D011837:Radiation-Protective Agents; C020174:N-(2-mercaptoethyl)-1,3-diaminopropane; D005978:Glutathione; D004999:Amifostine; C017807:bisbenzimide ethoxide trihydrochloride; D000781:Anethole Trithione