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Eun Chul Kim, Huan Meng, Mario Matthaei, Albert Jun; Viability of Bovine Corneal Endothelial Cells in Culture with Sulforaphane, VIP, or CNTF after Oxidative Stress. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6696.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the viability of bovine corneal endothelial cellsin culture with sulforaphane, Vasoactive intestinal peptide(VIP), or Ciliary neurotrophic factor (CNTF) after oxidativestress.
The bovine endothelial cells were cultured in Dulbecco's modifiedEagle's medium (DMEM) supplemented with 10% fetal bovine serum(FBS) in the absence (control) or presence of sulforaphane (5µM and 10 µM), VIP (10nM and 1µM), or CNTF(0.44 nM and 2.2 nM) for 48 hours. The cultures were then treatedwith 400 µM H2O2 for 30 minutes at 37°C in a humidified5% CO2 atmosphere. The cells were stained with calcein AM-ethidiumhomodimer, and analyzed with a fluorescence microscope. Experimentswere performed in triplicate for each drug concentration. Statisticalanalysis was performed using one-way ANOVA (SPSS version 16.0)and p<0.05 was statistically significant.
The percentages of dead cells in culture with sulforaphane 5µM (14.10 ± 4.05 %) and 10 µM (11.20 ±3.27 %), VIP 10nM (16.80 ± 3.67 %) and 1µM (22.31± 3.39 %), and CNTF 2.2 nM (23.55 ± 10.48 %) weresignificantly lower than that of control (35.11 ± 11.55%), respectively (P<0.05). The greatest reduction in percentdead cells was seen with sulforaphane 10 µM. Total cellcount of sulforaphane 5 µM (1331.50 ± 239.31 cell/mm2)and 10 µM (1205.66 ± 189.40 cell/mm2), and VIP10nM (1047.50 ± 208.45 cell/mm2) was significantly higherthan that of control (752.50 ± 335.12 cell/mm2), respectively(P<0.05).
Sulforaphane, VIP, and CNTF have a protective effect againstoxidative stress in bovine corneal endothelial cells. Theseagents may have potential as survival factors for endothelialcells under oxidative stress.
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