Investigating the putative therapeutic effects of sulfophane in the human lens
type: CreativeWork
description: Purpose: Cataract (a clouding of the lens) and PCO, a lens wound-healing response that occurs following cataract surgery, are important medical problems affecting millions worldwide. Reducing the incidence of these conditions would have a marked effect on the personal well being of millions of people. The isothiocyanate, SFN is known to have dose-dependent cytoprotective and cytotoxic properties. The present study had three aims. Firstly, to determine if the isothiocyanate, SFN could yield protection to lens cells against oxidative stress. Secondly, to identify the mechanisms by which SFN can elicit cytoprotection to lens cells against oxidative stress. The final aim was to establish the ability of SFN to initiate death of lens cells and prevent PCO formation. Methods: The human lens epithelial cell line FHL 124 was used in combination with whole porcine lens culture (to assess opacity) and human capsular bags (as a PCO model) were employed as the experimental systems. Whole lens cultures were monitored using brightfield and darkfield imaging; quantification was performed with ImageJ analysis software. Capsular bags were monitored using phase-contrast microscopy. The ApoToxGlo Triplex assay was used to assess FHL 124 cell survival, cytotoxicity and apoptosis. The MTS assay was used to assess cell populations. To determine levels of DNA strand breaks, the alkaline comet assay was performed and quantified. Lactate dehydrogenase levels in the medium (of FHL 124 cells and porcine lenses) were evaluated to reflect cell damage/death. To assess level of gene expression an Illumina whole genome HT-12 v4 beadchip microarray was employed. Real-time PCR was used to assess ER stress gene expression. Protein expression was validated by Western blot and immunocytochemistry (FHL 124 cells and capsular bags). Results: 30 M H2O2 exposure to FHL 124 cells caused a significant reduction in cell viability and increased cytotoxicity/apoptosis; these effects were significantly inhibited…
abstract: Purpose: Cataract (a clouding of the lens) and PCO, a lens wound-healing response that occurs following cataract surgery, are important medical problems affecting millions worldwide. Reducing the incidence of these conditions would have a marked effect on the personal well being of millions of people. The isothiocyanate, SFN is known to have dose-dependent cytoprotective and cytotoxic properties. The present study had three aims. Firstly, to determine if the isothiocyanate, SFN could yield protection to lens cells against oxidative stress. Secondly, to identify the mechanisms by which SFN can elicit cytoprotection to lens cells against oxidative stress. The final aim was to establish the ability of SFN to initiate death of lens cells and prevent PCO formation. Methods: The human lens epithelial cell line FHL 124 was used in combination with whole porcine lens culture (to assess opacity) and human capsular bags (as a PCO model) were employed as the experimental systems. Whole lens cultures were monitored using brightfield and darkfield imaging; quantification was performed with ImageJ analysis software. Capsular bags were monitored using phase-contrast microscopy. The ApoToxGlo Triplex assay was used to assess FHL 124 cell survival, cytotoxicity and apoptosis. The MTS assay was used to assess cell populations. To determine levels of DNA strand breaks, the alkaline comet assay was performed and quantified. Lactate dehydrogenase levels in the medium (of FHL 124 cells and porcine lenses) were evaluated to reflect cell damage/death. To assess level of gene expression an Illumina whole genome HT-12 v4 beadchip microarray was employed. Real-time PCR was used to assess ER stress gene expression. Protein expression was validated by Western blot and immunocytochemistry (FHL 124 cells and capsular bags). Results: 30 M H2O2 exposure to FHL 124 cells caused a significant reduction in cell viability and increased cytotoxicity/apoptosis; these effects were significantly inhibited…
author: Liu, Hanruo
language: en
date published: 2013-04
date modified: 2026-07-24T02:11:54Z
provider: University of East Anglia
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of East Anglia