Protective effects of carbenoxolone, an 11β-HSD1 inhibitor, against chemical induced dry eye syndrome
Dry eye syndrome (DES) is a disorder of the eye due to tear deficiency or excessive evaporation that causes damage to the eye and is associated with discomfort and dryness. 11β-Hydroxysteroid dehydrogenase 1 (11β-HSD1) is an enzyme that converts inactive cortisone to active cortisol. Recently, 11β-HSD1 has been expressed in human and rodent eyes and has been recognized as a target of glaucoma. In this study, the therapeutic effects and underlying mechanisms of topical carbenoxolone, an 11β-HSD1 inhibitor, were investigated in benzalkonium chloride (BAC)-treated human conjunctival epithelial cells and a rat DES model. In the in vitro study, carbenoxolone dose-dependently inhibited cell death and 11β-HSD1 activity in BAC-treated human conjunctival epithelial cells. For the in vivo study, carbenoxolone or a solvent was administered to the BAC-induced DES model twice daily. BAC-treated rat eyes showed significant increases in ocular surface damage, a reduction of tears, decrease corneal thickness, corneal basement membrane destruction, apoptosis in the conjunctival epithelium, and expression of pro-inflammatory cytokines (TNF-α and IL-6) and 11β-HSD1. These effects of BAC were reversed by topical carbenoxolone treatment. These results demonstrate that carbenoxolone can prevent DES by inhibiting pro-inflammatory cytokine expression and cell death of the corneal and conjunctival epithelium via inhibition of both 11β-HSD1 activity and expression in the eyes of BAC-treated rats. It is suggested that topical 11β-HSD1 inhibitors may provide a new therapeutic window in the prevention and/or treatment of DES.
KeywordsDry eye syndrome 11β-Hydroxysteroid dehydrogenase type 1 Carbenoxolone Benzalkonium chloride Apoptosis Inflammation
Dry eye syndrome
11β-Hydroxysteroid dehydrogenase 1
Tumor necrosis factor-α
Human conjunctival epithelial cells
Hematoxylin & Eosin
Periodic Acid & Schiff’s
- SD rat
Terminal deoxynucleotidyl transferase dUTP nick end labeling
We would like to acknowledge the financial support from the R&D Convergence Program (CCP-13-20-KRICT) and the DRC program (DRC-15-01-KRICT) of National Research Council of Science & Technology, and a project of the Korea Research Institute of Chemical Technology, the Ministry of Knowledge Economy, Republic of Korea.
Participated in research design: Y-JN, K-JC, and KYK. Conducted experiments: Y-JN, K-JC, SBP, H-RS, WHJ, and HYK. Contributed new reagents or analytic tools: HYK, and SDR. Performed data analysis: Y-JN, SDR, and KYK. Wrote or contributed to the writing of the manuscript: Y-JN, and KYK.
- 2.Pflugfelder SC (2008) Prevalence, burden, and pharmacoeconomics of dry eye disease. Am J Manag Care 14(3 Suppl):S102-106Google Scholar
- 5.Susarla R, Liu L, Walker EA, Bujalska IJ, Alsalem J, Williams GP, Sreekantam S, Taylor AE, Tallouzi M, Southworth HS, Murray PI, Wallace GR, Rauz S (2014) Cortisol biosynthesis in the human ocular surface innate immune response. PLoS ONE 9(4):e94913. doi: 10.1371/journal.pone.0094913 CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Bauskar A, Mack WJ, Mauris J, Argueso P, Heur M, Nagel BA, Kolar GR, Gleave ME, Nakamura T, Kinoshita S, Moradian-Oldak J, Panjwani N, Pflugfelder SC, Wilson MR, Fini ME, Jeong S (2015) Clusterin seals the ocular surface barrier in mouse dry eye. PLoS ONE 10(9):e0138958. doi: 10.1371/journal.pone.0138958 CrossRefPubMedPubMedCentralGoogle Scholar
- 15.Arumugam V, Vivekanandan K, Balasubramanian G, Kumar P, Sen R, S (2013) Role of 11β-hydorxysteroid dehydrogenase inhibitors in metabolic syndrome and its expansion in other therapeutic options. Int J Pharm research Bio-sci 2(5):78–99Google Scholar
- 23.Takamura E, Tsubota K, Watanabe H, Ohashi Y, Diquafosol Ophthalmic Solution Phase 3 Study G (2012) A randomised, double-masked comparison study of diquafosol versus sodium hyaluronate ophthalmic solutions in dry eye patients. Br J Ophthalmol 96 (10):1310–1315. doi: 10.1136/bjophthalmol-2011-301448 CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Rhee SD, Kim CH, Park JS, Jung WH, Park SB, Kim HY, Bae GH, Kim TJ, Kim KY (2012) Carbenoxolone prevents the development of fatty liver in C57BL/6-Lep ob/ob mice via the inhibition of sterol regulatory element binding protein-1c activity and apoptosis. Eur J Pharmacol 691(1–3):9–18. doi: 10.1016/j.ejphar.2012.06.021 CrossRefPubMedGoogle Scholar
- 31.Gilmour JS, Coutinho AE, Cailhier JF, Man TY, Clay M, Thomas G, Harris HJ, Mullins JJ, Seckl JR, Savill JS, Chapman KE (2006) Local amplification of glucocorticoids by 11 beta-hydroxysteroid dehydrogenase type 1 promotes macrophage phagocytosis of apoptotic leukocytes. J Immunol 176(12):7605–7611CrossRefPubMedGoogle Scholar
- 32.Chantong B, Kratschmar DV, Nashev LG, Balazs Z, Odermatt A (2012) Mineralocorticoid and glucocorticoid receptors differentially regulate NF-kappaB activity and pro-inflammatory cytokine production in murine BV-2 microglial cells. J Neuroinflammation 9:260. doi: 10.1186/1742-2094-9-260 CrossRefPubMedPubMedCentralGoogle Scholar
- 41.Moore JE, Vasey GT, Dartt DA, McGilligan VE, Atkinson SD, Grills C, Lamey PJ, Leccisotti A, Frazer DG, Moore TC (2011) Effect of tear hyperosmolarity and signs of clinical ocular surface pathology upon conjunctival goblet cell function in the human ocular surface. Invest Ophthalmol Vis Sci 52(9):6174–6180. doi: 10.1167/iovs.10-7022 CrossRefPubMedGoogle Scholar
- 47.Su X, Pradaux-Caggiano F, Thomas MP, Szeto MW, Halem HA, Culler MD, Vicker N, Potter BV (2010) Discovery of adamantyl ethanone derivatives as potent 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) inhibitors. ChemMedChem 5(7):1026–1044. doi: 10.1002/cmdc.201000081 CrossRefPubMedGoogle Scholar