Posters

Expression of the NLRP3 inflammasome in human corneal epithelial cells after desiccation in vitro

Poster Details

First Author: S.Kang SOUTH KOREA

Co Author(s):    A. Togloom   J. Oh   J. Song   H. Kim           

Abstract Details

Purpose:

To investigate whether desiccation induces NLRP3 inflammasome components expression in cultivated human corneal epithelial cells (HCEC).

Setting:

Department of Ophthalmology, Korea University College of Medicine, Seoul, Korea

Methods:

HCEC was grown in keratinocyte growth medium (K-SFM) to approximately 80% confluence. The medium was aspirated and dishes were left for 0 to 45 minutes with the cover left open to dry the cells. After desiccation, viability of the cells was estimated with CCK-8 assay. The expression of mRNA and protein levels of NLRP3 inflammasome components was measured by real-time PCR and western blot. Real-time PCR evaluated NLRP3, ASC and Caspase-1 mRNA levels. Western blot analysis assessed NLRP3, ASC, pro-caspase-1 and active caspase-1 protein expression.

Results:

Desiccation changed the cell morphology and decreased the viability of HCEC. The mRNA expression of NLRP3 was increased in a dose-dependent manner with desiccation, but that of ASC and caspase-1 was not increased. Protein levels of NLRP3 and conversion of pro-caspase-1 into active caspase-1 were significantly increased after desiccation.

Conclusions:

Desiccation induced NLRP3 inflammasome expression in human corneal epithelial cells, suggesting that inflammasome may participates in desiccation-induced inflammatory process. NLRP3 inflammasome might play a priming role in the pathogenesis of dry eye disease.

Financial Disclosure:

None