Cytotoxic effect and possible mechanisms of Tetracaine on human corneal epithelial cells in vitro
Cytotoxic effect and possible mechanisms of Tetracaine on human corneal epithelial cells in vitro作者机构：Key Laboratory for Corneal Tissue Engineering College of Marine Life Sciences Ocean University of China
出 版 物：《International Journal of Ophthalmology(English edition)》 (国际眼科杂志（英文版）)
年 卷 期：2016年第9卷第4期
页 面：497-504页
核心收录：
学科分类：1002[医学-临床医学] 100212[医学-眼科学] 10[医学]
主 题：Tetracaine cytotoxicity human cornel epithelial cells apoptosis mitochondrion
摘 要：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, phosphati-dylserine 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 from10.0 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 phosphati-dylserine externalization, DNA fragmentation, G1 phase arrest, and ultrastructural abnormality and apoptotic body formation. Furthermore, Tetracaine at a dose of0.3125 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-x L.·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 cellapoptosis via a death receptor-mediated mitochondriondependent pathway.
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