April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Increased Substance P Expression in the Ocular Surface in Murine Dry Eye Disease
Author Affiliations & Notes
  • Sang-Mok Lee
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
    Ophthalmology, Hallym University Sacred Heart Hospital, Anyang, Republic of Korea
  • Zahra Sadrai
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Hyun Soo Lee
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
    Ophthalmology and Visual Science, The Catholic University of Korea School of Medicine, Seoul, Republic of Korea
  • William Stevenson
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Yihe Chen
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Jing Hua
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Kishore Reddy Katikireddy
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Thomas H Dohlman
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Sunil K Chauhan
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Reza Dana
    Dana lab, Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA
  • Footnotes
    Commercial Relationships Sang-Mok Lee, None; Zahra Sadrai, None; Hyun Soo Lee, None; William Stevenson, None; Yihe Chen, None; Jing Hua, None; Kishore Reddy Katikireddy, None; Thomas Dohlman, None; Sunil Chauhan, None; Reza Dana, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 3664. doi:https://doi.org/
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      Sang-Mok Lee, Zahra Sadrai, Hyun Soo Lee, William Stevenson, Yihe Chen, Jing Hua, Kishore Reddy Katikireddy, Thomas H Dohlman, Sunil K Chauhan, Reza Dana; Increased Substance P Expression in the Ocular Surface in Murine Dry Eye Disease. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3664. doi: https://doi.org/.

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

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Abstract

Purpose: Substance P is a key mediator of pain and neurogenic inflammation. In the present study, we analyzed protein and mRNA expression levels of substance P in the ocular surface to evaluate their association with the early induction phase of dry eye disease (DED).

Methods: DED was induced in 6-week-old female C57BL/6 mice by bilateral extra-orbital lacrimal gland excision followed by their placement in a controlled environmental chamber. Corneal fluorescent staining (CFS) was evaluated at days -1, 3, 6, and 13, and corneas and conjunctivae were harvested at days 4, 7, and 14 to analyze protein and mRNA expression levels of substance P. Substance P protein expression levels were measured with a competitive enzyme immunoassay kit (R&D Systems, Minneapolis, MN) and then adjusted to the total protein amount. Expression of substance P mRNA was analyzed by real-time PCR. One way ANOVA test was used for comparison of the expression levels and Pearson correlation was used for correlation analysis.

Results: Substance P protein expression levels were increased in both the cornea and conjunctiva on days 4 and 7 compared to the sham operated control group (Cornea: 1.9 fold on day 4, 2.0 fold on day 7, P<0.05; Conjunctiva: 2.8 fold on day 4, 2.2 fold on day 7, P<0.05). By day 14, levels of substance P decreased to baseline levels. We observed similar kinetics for the mRNA expression of substance P. Increased protein expression of substance P in the cornea correlated with increased CFS scores until day 7 (r = 0.900, P<0.001).

Conclusions: Protein and mRNA expression levels of substance P are increased in the early induction phase of DED and correlate with increased CFS scores.

Keywords: 480 cornea: basic science • 614 neuropeptides • 533 gene/expression  
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