Abstract
Purpose :
Topical spironolactone has been found to be effective at treating dry eye disease (Yee, 2016, 2017), and a patent has been issued to one of the authors (RY). The purpose of this study was to begin to elucidate physiological mechanisms for the successful use of spironolactone for dry eye disease.
Methods :
The influence of spironolactone on primary human corneal epithelial cell (HCE) lipid production, fibrotic- and antifibrotic-associated proteins, migration and proliferation were examined. Lipid production was examined using Oil Red O staining, qPCR measured mRNA levels of PPARγ, and TGF β1 and β3. Migration was examined using a scratch assay, and proliferation was studied using the MTT assay.
Results :
HCE Oil Red O staining was scored as more intense for all trials (0.005, 0.01, 0.02 mg/ml for 4 or 6 days). The highest concentration and longest durations received the highest scores. qPCR was performed on HCE from 3 different donors using the same concentrations in cells exposed to spironolactone for 24, 48, and 72 h. TGF β3 mRNA levels were significantly increased in 2 of 3 sets of cells in a dose but not time-dependent manner. TGF β1 mRNA was increased in all samples except 1 concentration/time point, where it was decreased. PPARγ mRNA levels both increased and decreased depending on the donor, although it decreased in at least one concentration/time point in all donor samples tested. HCE treated for 48 hours with spironolactone (0.01 mg/ml) showed significantly increased migration and inhibited proliferation at all time points.
Conclusions :
We hypothesize that spironolactone is in part beneficial for dry eye treatment due to its possible effects on corneal epithelium. Dry eye syndrome is characterized in part by epithelial defects. We found increased spironolactone-induced HCE lipid production and increased TGF β3 mRNA levels in spironolactone treated cells. TGF β3 has been found to have anti-fibrotic activity in the cornea (Karamichos et al, 2014). Spironolactone also increased HCE migration. It had no effect on PPARγ mRNA, which has been associated with meibocyte differentiation and lipid synthesis (Jester et al, 2018). On the other hand, spironolactone increased HCE TGF β1 mRNA , which is associated with fibrosis, and inhibited proliferation. We conclude that the beneficial effects of spironolactone on HCE are in part responsible for its successful use of in treating dry eye disease.
This is a 2020 ARVO Annual Meeting abstract.