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Cytotoxic effect and possible mechanisms of Tetracaine on human corneal epithelial cells in vitro
- Source :
- International Journal of Ophthalmology, Vol 9, Iss 4, Pp 497-504 (2016)
- Publication Year :
- 2016
- Publisher :
- Press of International Journal of Ophthalmology (IJO Press), 2016.
-
Abstract
- AIM: To demonstrate the cytotoxic effect and possible mechanisms of Tetracaine on human corneal epithelial (HCEP) cells in vitro. METHODS: In vitro cultured HCEP cell were treated with Tetracaine hydrochloride at different doses for different times, and their morphology, viability, and plasma membrane permeability were detected by light microscopy, methyl thiazolyl tetrazolium (MTT) assay, and acridine orange (AO)/ethidium bromide (EB) staining, respectively. Their cell cycle progression, phosphatidylserine orientation in plasma membrane, and mitochondrial membrane potential (MTP) were assessed by flow cytometry. DNA fragmentation, ultrastructure, caspase activation, and the cytoplasmic apoptosis inducing factor (AIF) and cytochrome c (Cyt. c) along with the expression of B-cell lymphoma-2 (Bcl-2) family proteins were examined by gel electrophoresis, transmission electron microscope, enzyme linked immunosorbent assay (ELISA), and Western blot, respectively. RESULTS: After exposed to Tetracaine at doses from 10.0 to 0.3125 g/L, the HCEP cells showed dose- and time-dependent morphological abnormality and typical cytopathic effect, viability decline, and plasma membrane permeability elevation. Tetracaine induced phosphatidylserine externalization, DNA fragmentation, G1 phase arrest, and ultrastructural abnormality and apoptotic body formation. Furthermore, Tetracaine at a dose of 0.3125 g/L also induced caspase-3, -9 and -8 activation, MTP disruption, up-regulation of the cytoplasmic amount of Cyt. c and AIF, the expressions of Bax and Bad, and down-regulation of the expressions of Bcl-2 and Bcl-xL. CONCLUSION: Tetracaine above 0.3125 g/L (1/32 of its clinical applied dosage) has a dose- and time-dependent cytotoxicity to HCEP cells in vitro, with inducing cell apoptosis via a death receptor-mediated mitochondrion-dependent pathway.
- Subjects :
- Tetracaine
human cornel epithelial cells
010501 environmental sciences
Mitochondrion
01 natural sciences
03 medical and health sciences
0302 clinical medicine
lcsh:Ophthalmology
medicine
mitochondrion
Cytotoxic T cell
Cytotoxicity
0105 earth and related environmental sciences
business.industry
apoptosis
eye diseases
In vitro
Ophthalmology
Basic Research
lcsh:RE1-994
Apoptosis
Immunology
030221 ophthalmology & optometry
Cancer research
cytotoxicity
sense organs
business
medicine.drug
Subjects
Details
- ISSN :
- 22274898 and 22223959
- Database :
- OpenAIRE
- Journal :
- International Journal of Ophthalmology
- Accession number :
- edsair.doi.dedup.....21c15cf97faafa7d9952c4ade7f41a04