Abstract
Oxidative damage is associated with multiple ocular diseases including corneal disorders, cataract, and retinal degeneration. Thus, the possibility to use the multifunctional antioxidant carcinine (β-alanyl-histamine) as a therapeutic to prevent and reverse intracellular oxidative damage in ocular diseases is promising. Today, the major barrier to developing new ophthalmic treatments is the identification and implementation of effective methods of drug delivery in the eye. The aim of this study was to investigate carcinine transport and the expression profile of transporter genes in corneal epithelial cells. These cells form the blood-aqueous barrier of the cornea, preventing the penetration of topical drugs.
The posterior tissue bioavailability and biological effect of carcinine was measured in mouse retina, following topical delivery. The transport and biological effect of carcinine was measured in cultured human corneal epithelial cells (HCECs). The expression profile of 84 transporter genes was determined in HCECs by quantitative RT-PCR.
We found that carcinine efficiently penetrated the eye globe and the retinal cells after topical delivery in mouse eyes. However, carcinine was not transported into the intracellular compartment of the HCECs and the putative carcinine transporters PepT1 and PepT2 had negligible expression levels in these cells. Five other members of the solute carrier (SLC) superfamily of transporters, encoding influx pumps for amino acids, monocarboxylates, and copper ions were highly expressed in HCECs.
We conclude that carcinine does not penetrate the eye globe through the blood-aqueous barrier of the cornea, suggesting a conjunctival/scleral penetration. Identification of transporters SLC3A2, SLC7A5, SLC16A1, SLC31A1, and SLC38A2 as highly expressed in the corneal epithelium may facilitate the design of topical ophthalmic drugs with enhanced intraocular bioavailability.
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Abbreviations
- 4-HNE:
-
4-Hydroxynonenal
- ABC:
-
ATP-binding cassette
- ATA2:
-
Amino acid transporter A2
- ATP:
-
Adenosine triphosphase
- COPT1:
-
Copper transporter 1
- Ct:
-
Cycle threshold
- HCEC:
-
Human corneal epithelial cell
- HPLC:
-
High-performance liquid chromatography
- HPRT1:
-
Hypoxanthine phosphoribosyltransferase
- LAT1:
-
Large neutral amino acid transporter 1
- MCT1:
-
Monocarboxylate transporter 1
- MRP:
-
Multidrug resistance-associated protein
- MS:
-
Mass spectrometry
- PEPT:
-
Peptide transporter
- PMP70:
-
Peroxisomal membrane protein of 70 kDA
- RT-PCR:
-
Reverse transcription polymerase chain reaction
- SLC:
-
Solute carrier
- VDAC:
-
Voltage-dependent anion channel
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Acknowledgements
This work was supported by grants HR10-152 from the Oklahoma Center for the Advancement of Science and Technology and R01EY015534 from the National Institute of Health. We would like to thank the Laboratory for Molecular Biology and Cytometry Research at the University of Oklahoma Health Sciences Center for the HPLC-MS analysis of unlabeled carcinine.
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Kasus-Jacobi, A., Nag, O.K., Awasthi, V., Babizhayev, M.A., Pereira, H.A. (2015). Investigating Carcinine Transport and the Expression Profile of Transporter Genes in Human Corneal Epithelial Cells. In: Babizhayev, M., Li, DC., Kasus-Jacobi, A., Žorić, L., Alió, J. (eds) Studies on the Cornea and Lens. Oxidative Stress in Applied Basic Research and Clinical Practice. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1935-2_8
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DOI: https://doi.org/10.1007/978-1-4939-1935-2_8
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