Cytotoxicity of carteolol to human corneal epithelial cells by inducing apoptosis via triggering the Bcl-2 family protein-mediated mitochondrial pro-apoptotic pathway. (September 2016)

• Record Type: Journal Article
• Authors: Shan, Ming; Fan, Ting-Jun
• Is Part Of: Toxicology in vitro. Volume 35(2016)
• Page Start: 36
• Page End: 42
• Publication Date: 2016-09
• Subjects: AIF apoptosis inducing factor -- ANOVA one-way analysis of variance -- AO/EB acridine orange/ethidium bromide -- CPE cytopathic effect -- Cyt. c cytochrome c -- DAB 3, 3′N-diaminobenzidine tertrahydrochloride -- FBS fetal bovine serum -- FCM flow cytometry -- HCEP human corneal epithelium -- JC-1 5, 5′, 6, 6′-tetrachloro-1.1′, 3, 3′-tetraethybenzimida -- MTP mitochondrial transmembrane potential -- MTT methyl thiazolyl tetrazolium -- PI propidium iodide -- PM plasma membrane -- PMSF phenylmethylsulfonyl fluoride -- PS phosphatidylserine -- SDS-PAGE SDS-polyacrylamide gel electrophoresis -- TEM transmission electron microscopy -- TMB tetramethylbenzidine; Carteolol -- Human corneal epithelial cells -- Cytotoxicity -- Apoptosis -- Mitochondrion; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2016.05.008
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Carteolol is a frequently used nonselective β-adrenoceptor antagonist for glaucoma and ocular hypertension treatment, and its repeated/prolonged usage might be cytotoxic to the cornea, especially the outmost human corneal epithelium (HCEP). The aim of the present study was to characterize the cytotoxicity of carteolol to HCEP and its underlying cellular and molecular mechanisms using an in vitro model of HCEP cells. After HCEP cells were treated with carteolol at concentrations varying from 2% to 0.015625%, the cytotoxicity, apoptosis-inducing effect and pro-apoptotic pathway was investigated, respectively. Our results showed that carteolol at concentrations above 0.03125% induced time- and dose-dependent growth retardation, cytopathic morphological changes and viability decline of HCEP cells. Moreover, carteolol induced G1 phase arrest, plasma membrane permeability elevation, phosphatidylserine externalization, DNA fragmentation, and apoptotic body formation of HCEP cells. Furthermore, carteolol also induced activation of caspase-9 and − 3, disruption of mitochondrial transmembrane potential, up-regulation the cytoplasmic amount of cytochrome c and apoptosis-inducing factor, and up-regulation of pro-apoptotic Bax and Bad, down-regulation of anti-apoptotic Bcl-2 and Bcl-xL. In conclusion, carteolol above 1/64 of its clinical therapeutic dosage has a time- and dose-dependent cytotoxicity to HCEP cells, which is achieved by inducing apoptosis via triggering Bcl-2 family protein-mediated mitochondrial pro-apoptotic pathway. Graphical abstract: Highlights: Carteolol has cytotoxicity to human corneal epithelial cells in vitro . The cytotoxicity of carteolol is dose- and time-dependent. Carteolol induces G1-phase arrest and apoptosis of these cells. The carteolol-induced apoptosis is mediated by Bcl-2 family proteins. The pro-apoptotic pathway of carteolol is mitochondria-dependent.