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M Matsunaga, Y Saitoh, H Nakashima, S Nakamura, F Saito, H Aburatani, A Higuchi, K Tsubota; Effect of D-3-Hydroxybutyrate (HBA) on Apoptosis Induced by Serum Deprivation in Cultured Conjunctival Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3161.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:Previous studies have shown that HBA, which is one of endogenous substrates for energy, promotes neuronal survival in model of Alzheimer's disease, reduces cerebral edema formation in ischemic brain, and suppress posttraumatic protein catabolism. We presented that topical HBA shows a protective effect against corneal and conjunctival disorders on rats (ARVO 2000) . In this study we evaluated the effect of HBA on apoptosis induced by serum deprivation in cultured conjunctival epithelial cells. Methods: Conjunctival epithelial cell line, CCL20.2 (CCL) was grown in Medium 199 with 10% fetal bovine serum (FBS) for 3 days, and the medium was prepared as Medium 199 with 0 to 80mM HBA, with 0 to 10 % FBS, or with 80mM L-3-Hydroxybutyrate, a optical isomer of HBA. After 0.5, 1, 3, 6, 12 and 24hrs incubation, we observed cell formation, measured cell viability, and measured caspase-3 activity in each cell lysate.Results: Serum deprivation caused immediate caspase-3 activation, cell detachment after 3hrs, and cell viability decreased to 34.9% for 24hrs. HBA and FBS suppressed caspase-3 activation and cell detachment in a does dependent manner. Cell viabilities, at 24hrs in 80mM HBA and 10% FBS were 71.6% and 97.4%, respectively. The optical isomer of HBA showed no effect.Conclusion: These results indicate that HBA rescues CCL from cell death of serum deprivation by way of suppressing apoptotic activity, therefore topical HBA would relieves ocular surface disorder.
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