Abstract
Purpose. The purpose of this study was to determine whether 25-hydroxyvitamin D 3 (25(OH)D 3) and/or its active metabolite, 1α,25-dihydroxyvitamin D 3 (1,25(OH) 2D 3), can enhance corneal epithelial barrier function. The authors also determined if corneas contain mRNA for the vitamin D receptor (VDR) and 1α-hydroxylase, the enzyme required to convert 25(OH)D 3 to 1,25(OH)2D 3, and measured vitamin D metabolite concentrations in aqueous and vitreous humor. Methods. RT-PCR was used to examine mouse, rabbit, and human corneal epithelial VDR and 1α-hydroxylase mRNA. Vitamin D metabolites were measured using a selective vitamin D derivatizing agent and mass spectroscopy. Barrier function experiments were performed by measuring inulin permeability (IP) and/or transepithelial resistance (TER) in control, 25(OH)D 3-, and 1,25(OH)2D 3-treated human and rabbit corneal epithelial monolayers cultured on permeable inserts. Ca 2+ was removed, then reintroduced to the culture medium while IP and TER readings were taken. Occludin levels were examined using Western blotting. Results. All corneal samples were positive for both VDR and 1α-hydroxylase mRNA. All vitamin D metabolites except for unhydroxylated vitamin D 3 were detected in aqueous and vitreous humor. Epithelial cells showed increased TER, decreased IP, and increased occludin levels when cultured with 25(OH)D 3 and 1,25(OH) 2D 3. Conclusions. We conclude that corneas contain mRNA for VDR and 1α-hydroxylase as well as significant vitamin D concentrations. 25(OH)D 3 and its active metabolite 1,25(OH) 2D 3, both enhance corneal epithelial barrier function.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 7359-7364 |
| Number of pages | 6 |
| Journal | Investigative Ophthalmology and Visual Science |
| Volume | 52 |
| Issue number | 10 |
| DOIs | |
| State | Published - Sep 1 2011 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Ophthalmology
- Sensory Systems
- Cellular and Molecular Neuroscience
- Medicine(all)
Cite this
Vitamin D enhances corneal epithelial barrier function. / Yin, Zhaohong; Pintea, Victorina; Lin, Yanping; Hammock, Bruce D.; Watsky, Mitchell Aaron.
In: Investigative Ophthalmology and Visual Science, Vol. 52, No. 10, 01.09.2011, p. 7359-7364.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Vitamin D enhances corneal epithelial barrier function
AU - Yin, Zhaohong
AU - Pintea, Victorina
AU - Lin, Yanping
AU - Hammock, Bruce D.
AU - Watsky, Mitchell Aaron
PY - 2011/9/1
Y1 - 2011/9/1
N2 - Purpose. The purpose of this study was to determine whether 25-hydroxyvitamin D 3 (25(OH)D 3) and/or its active metabolite, 1α,25-dihydroxyvitamin D 3 (1,25(OH) 2D 3), can enhance corneal epithelial barrier function. The authors also determined if corneas contain mRNA for the vitamin D receptor (VDR) and 1α-hydroxylase, the enzyme required to convert 25(OH)D 3 to 1,25(OH)2D 3, and measured vitamin D metabolite concentrations in aqueous and vitreous humor. Methods. RT-PCR was used to examine mouse, rabbit, and human corneal epithelial VDR and 1α-hydroxylase mRNA. Vitamin D metabolites were measured using a selective vitamin D derivatizing agent and mass spectroscopy. Barrier function experiments were performed by measuring inulin permeability (IP) and/or transepithelial resistance (TER) in control, 25(OH)D 3-, and 1,25(OH)2D 3-treated human and rabbit corneal epithelial monolayers cultured on permeable inserts. Ca 2+ was removed, then reintroduced to the culture medium while IP and TER readings were taken. Occludin levels were examined using Western blotting. Results. All corneal samples were positive for both VDR and 1α-hydroxylase mRNA. All vitamin D metabolites except for unhydroxylated vitamin D 3 were detected in aqueous and vitreous humor. Epithelial cells showed increased TER, decreased IP, and increased occludin levels when cultured with 25(OH)D 3 and 1,25(OH) 2D 3. Conclusions. We conclude that corneas contain mRNA for VDR and 1α-hydroxylase as well as significant vitamin D concentrations. 25(OH)D 3 and its active metabolite 1,25(OH) 2D 3, both enhance corneal epithelial barrier function.
AB - Purpose. The purpose of this study was to determine whether 25-hydroxyvitamin D 3 (25(OH)D 3) and/or its active metabolite, 1α,25-dihydroxyvitamin D 3 (1,25(OH) 2D 3), can enhance corneal epithelial barrier function. The authors also determined if corneas contain mRNA for the vitamin D receptor (VDR) and 1α-hydroxylase, the enzyme required to convert 25(OH)D 3 to 1,25(OH)2D 3, and measured vitamin D metabolite concentrations in aqueous and vitreous humor. Methods. RT-PCR was used to examine mouse, rabbit, and human corneal epithelial VDR and 1α-hydroxylase mRNA. Vitamin D metabolites were measured using a selective vitamin D derivatizing agent and mass spectroscopy. Barrier function experiments were performed by measuring inulin permeability (IP) and/or transepithelial resistance (TER) in control, 25(OH)D 3-, and 1,25(OH)2D 3-treated human and rabbit corneal epithelial monolayers cultured on permeable inserts. Ca 2+ was removed, then reintroduced to the culture medium while IP and TER readings were taken. Occludin levels were examined using Western blotting. Results. All corneal samples were positive for both VDR and 1α-hydroxylase mRNA. All vitamin D metabolites except for unhydroxylated vitamin D 3 were detected in aqueous and vitreous humor. Epithelial cells showed increased TER, decreased IP, and increased occludin levels when cultured with 25(OH)D 3 and 1,25(OH) 2D 3. Conclusions. We conclude that corneas contain mRNA for VDR and 1α-hydroxylase as well as significant vitamin D concentrations. 25(OH)D 3 and its active metabolite 1,25(OH) 2D 3, both enhance corneal epithelial barrier function.
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U2 - 10.1167/iovs.11-7605
DO - 10.1167/iovs.11-7605
M3 - Article
VL - 52
SP - 7359
EP - 7364
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
SN - 0146-0404
IS - 10
ER -