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Hanruo Liu, Andrew J. Smith, Martin C. Lott, Yongping Bao, Richard P. Bowater, John R. Reddan, I M. Wormstone; Sulforaphane Can Protect Human Lens Cells Against Oxidative Stress. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1066.
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© ARVO (1962-2015); The Authors (2016-present)
Protecting the lens against oxidative stress is of great importance in delaying the onset of cataract. Isothiocyanates such as sulforaphane (SFN) that can be found in food sources that include broccoli and cauliflower have been proposed to provide cytoptotection against oxidative stress. We therefore tested the ability of sulforaphane to perform this role in lens cells.
The human lens epithelial cell line FHL124 was used. The ApoToxGlo assay (Promega) was used to assess cell survival, cytotoxicity and apoptosis. Lactose dehydrogenase levels in the medium were evaluated to reflect cell damage/death and the TUNEL assay was used to determine apoptosis. To determine single and double strand break DNA damage the alkaline comet assay was performed and quantified using Comet IV analysis software (Perceptive Instruments). To assess the levels of gene expression an Illumina whole genome HT-12 v4 beadchip was employed and analysed using the Bioconductor package in R.
Using the ApoToxGlo assay, cell viability, cytoxicity and apoptosis levels did not differ from non-treated control at concentrations ≤5μM. At 10μM and above a progressive loss of viability and increased cytotoxicity/apoptosis were seen. Application of 30μM H2O2 to FHL124 cells caused a reduction in cell viability and increased cytotoxicity/apoptosis; these effects were significantly inhibited by 24h pre-treatment with 1μM SFN. 30μM H2O2 also caused an elevation in LDH levels in the medium relative to control, which were suppressed by SFN treatment. A similar pattern was observed with the TUNEL assay. DNA damage was significantly increased within 30 mins of exposure to 30μM H2O2, whereas, levels in the 1μM SFN group did not significantly differ from controls. Pre-treatment with 1μM SFN significantly reduced H2O2-induced DNA damage. Gene microarray data revealed a number of genes up-regulated following 24 hour exposure to 1μM SFN, which included NQO1, TXNRD1, G6PD, EPHX and GCLM.
The dietary component, SFN, demonstrates an ability to protect human lens cells against oxidative stress and thus could potentially delay the onset of cataract
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