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Suneel Gupta, Nishant Rajiv Sinha, Praveen Balne, Ratnakar Tripathi, Nathan Hesemann, Prashant R. Sinha, Rajiv R Mohan; Mechanism associated with acrolein’s toxicity to the cornea in vitro. Invest. Ophthalmol. Vis. Sci. 2021;62(8):942.
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Acrolein, a volatile organic compound, was used as a warfare agent in World War I and has the potential viable public threat worldwide. Recently, we studied the impact of acrolein exposure to the cornea and other ocular surface tissues in vivo using a rabbit model. The study sought to uncover the mechanisms of acrolein’s toxicity to the cornea using an in vitro model.<!--[if !supportLineBreakNewLine]--><!--[endif]--><!--[if !supportLineBreakNewLine]--><!--[endif]-->
Primary human corneal stromal fibroblasts cultures (h-CSFs) from donor human corneas were used. Presto blue and MTT assays were used to determine the dose of acrolein, N-acetyl cysteine (NAC), and Buthionine sulphoximine (BSO) for h-CSFs. Different biochemical assays using standard commercial kits and qRT-PCR analysis were performed to understand the mode of acrolein toxicity.
The MTT and PrestoBlue assays defined that 100 µM dose of acrolein represents IC50, for h-CSFs an acute level at 4h (p<0.001). The biochemical assays revealed that acrolein exposure to h-CSFs led to oxidative stress and a compromised level of glutathione. These biochemical variations led to mitochondrial dysfunction via changes in the mitochondrial membrane potential (ΔΨm). The mRNA data, TUNEL, and caspase-3/7 assays showed that acrolein induces apoptosis in h-CSFs. Treatment with NAC; a precursor of glutathione, in h-CSFs decreased the oxidative stress, elevated the GSH level, regulated the mitochondrial membrane potential (ΔΨm), and rescued h-CSFs from apoptosis under acrolein-exposed conditions.<!--[if !supportLineBreakNewLine]--><!--[endif]-->
Acrolein’s exposure to human corneal fibroblasts induced alterations in phenotype, and cell death through oxidative damage at least in part of toxicity, in vitro.
This is a 2021 ARVO Annual Meeting abstract.
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