July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Histatin 5 can reduce apoptosis induced by hyperosmolar conditions in human corneal epithelial cells.
Author Affiliations & Notes
  • Vinay Kumar Aakalu
    Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, Chicago, Illinois, United States
    Surgery, Jesse Brown Veterans Affairs Hospital, Chicago, Illinois, United States
  • Dhara Shah
    Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, Chicago, Illinois, United States
  • Sushma Kalmodia
    Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, Chicago, Illinois, United States
  • Marwan Ali
    Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, Chicago, Illinois, United States
  • Arun Balasubramaniam
    Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, Chicago, Illinois, United States
  • Kyung-No Son
    Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Vinay Aakalu, None; Dhara Shah, None; Sushma Kalmodia, None; Marwan Ali, None; Arun Balasubramaniam, None; Kyung-No Son, None
  • Footnotes
    Support  NEI/NIH K08EY024339, Department of Defense:W81XWH1710122, VA I01BX004080, Unrestricted Grant, Research to Prevent Blindness NEI P30EY001792
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 930. doi:
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      Vinay Kumar Aakalu, Dhara Shah, Sushma Kalmodia, Marwan Ali, Arun Balasubramaniam, Kyung-No Son; Histatin 5 can reduce apoptosis induced by hyperosmolar conditions in human corneal epithelial cells.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):930.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Hyperosmolar-induced ocular surface cell death is a mitochondria-mediated event in inflammatory eye diseases such as dry eye. Hyperosmolarity is a key element of dry eye disease pathophysiology. It has been reported that hyperosmolarity induces apoptosis of human corneal epithelial cells through a cytochrome c-mediated death pathway, which may be dependent upon, or modulated by, JNK and ERK MAPK signaling pathways. Histatin peptides are well established antimicrobial and wound healing agents and have minimal to no immunogenicity or side effects. The mechanisms of action of histatin peptides are poorly understood. We studied the role of Histain 5 (H5) and its involvement in hyperosmolarity-stimulated apoptotic cell death in human corneal epithelial (HCE) cells.

Methods : HCE cells were stimulated in hyperosmolar media (450 mOsM) by adding 100 mM NaCl, with or without H5 (20 µM). For measuring apoptotic activity levels, we checked cleavage of caspase-3 and PARP-1 with a Western blot assay. For quantitative measurements, caspase-3 activity was assayed with the Caspase-Glo reagent (Promega, Madison, WI) in HCE cells stimulated with Hosm, and treated with H5. Additionally, the activity of p38, JNK and ERK were analyzed using phospho-specific antibodies.

Results : H5 inhibited the apoptosis stimulated by hyperosmolar solution (450 mOsM) in HCE cells. Cleavage of active caspase, caspase-3 and PARP-1 was observed in hyperosmolar solution in HCE cells, the treatment of H5 inhibited their activation at 8 and 24 hrs. Quantification of caspase-3 activity revealed significant inhibition of caspase-3 after co- treatment with H5 compared to treatment with hyperosmolar solution alone at 16 hr. in HCE cells. Phosphorylation of p38 and JNK1/2 MAPKs was inhibited at 30 min after treatment with H5; however, Pro-survival AKT and MAPK-ERK1/2 proteins were activated in HCE cells.

Conclusions : H5 can reduce apoptosis induced by hyperosmolar solution in HCE cells. Activation of survival pathways and reduction of activity of apoptotic pathways was noted with H5 exposure. Taken together these results suggest that H5 may have utility in reducing corneal epithelial cell apoptosis and enhancing survival in hyperosmolar solutions.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

Cells were stimulated in hyperosmolar media (450 mOsM) with or without H5 (20 µM). Notable is significant induction of caspase3/7 at 16hrs by HOsm and reduction by H5 co-treatrment.

Cells were stimulated in hyperosmolar media (450 mOsM) with or without H5 (20 µM). Notable is significant induction of caspase3/7 at 16hrs by HOsm and reduction by H5 co-treatrment.

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