June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
L-carnitine, Erythritol and Betaine Suppress the Production and Activity of Matrix Metalloproteinases in Primary Human Corneal Epithelial Cells Exposed to Hyperosmotic Stress
Author Affiliations & Notes
  • Ruzhi Deng
    Department of Ophthalmology, Baylor College of Medicine, Houston, TX
    Optometry and Ophthalmology, Wenzhou Medical College, Wenzhou, China
  • Zhitao Su
    Department of Ophthalmology, Baylor College of Medicine, Houston, TX
    Optometry and Ophthalmology, Wenzhou Medical College, Wenzhou, China
  • Jing Lin
    Department of Ophthalmology, Baylor College of Medicine, Houston, TX
  • Xia Hua
    Department of Ophthalmology, Baylor College of Medicine, Houston, TX
  • De-Quan Li
    Department of Ophthalmology, Baylor College of Medicine, Houston, TX
  • Stephen Pflugfelder
    Department of Ophthalmology, Baylor College of Medicine, Houston, TX
  • Footnotes
    Commercial Relationships Ruzhi Deng, None; Zhitao Su, None; Jing Lin, None; Xia Hua, None; De-Quan Li, None; Stephen Pflugfelder, Allergan (C), Glaxo Smith Kline (C), Bausch and Lomb (C), Baylor College of Medicine (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 4315. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Ruzhi Deng, Zhitao Su, Jing Lin, Xia Hua, De-Quan Li, Stephen Pflugfelder; L-carnitine, Erythritol and Betaine Suppress the Production and Activity of Matrix Metalloproteinases in Primary Human Corneal Epithelial Cells Exposed to Hyperosmotic Stress. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4315.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: Hyperosmolarity has been recognized to be a pro-inflammatory stress in the pathogenesis of dry eye disease. This study investigated the suppressive effect of osmoprotectants (L-carnitine, erythritol and betaine) on the production and activity of matrix metalloproteinases (MMPs) in primary human corneal epithelial cells (HCECs) exposed to hyperosmotic stress.

Methods: Primary HCECs were established from fresh donor limbal tissue explants. The cultures in iso-osmolar medium (312 mOsM) were switched to hyperosmotic media (400-500 mOsM) by adding 50-90 mM NaCl, with or without prior incubation with different concentrations (2, 10 or 20mM) of L-carnitine, erythritol or betaine. The mRNA expression of MMPs by HCECs treated for 4 hours was determined by reverse transcription and quantitative real time PCR. Their protein production and activity in the conditioned media from cultures treated for 24-48 hours were evaluated by zymography, immunofluorescent staining, ELISA and activity assays.

Results: Hyperosmotic stress (400, 450 or 500 mOsM) significantly stimulated the mRNA expression of collagenase MMP13 (3.1 to 7.5 fold), gelatinase MMP9 (1.7 to 2.5 fold), stromelysin MMP3 (2.2 to 4.1 fold) and matrilysin MMP7 (2.3 to 4.5 fold), mostly in an osmolarity dependent fashion. Interestingly, the stimulated expression of these MMPs was significantly, but differentially suppressed by L-carnitine, erythritol or betaine. L-carnitine appeared to have the greatest inhibitory effects, and down-regulated 52-78%, respectively, of the stimulated mRNA levels of MMP13 (down from 7.0 to 3.1 fold), MMP9 (2.5 to 1.2 fold), MMP3 (4.1 to 0.9 fold), and MMP7 (4.5 to 1.5 fold) by HCECs exposed to 450 mOsM. The stimulated production and activity of these MMPs by hyperosmotic stress and the suppressive effects of L-carnitine, erythritol and betaine were further confirmed at protein levels by gelatin and casein gel zymography, immunofluorescent staining, ELISA and/or activity assays, respectively.

Conclusions: Our findings demonstrate that hyperosmotic stress stimulates the expression, production and activity of MMPs in HCECs. L-carnitine, erythritol and betaine serve as osmoprotectants that suppress the production and activity of MMPs in HCECs exposed to hyperosmotic stress. L-carnitine shows the best inhibitory effect among the three.

Keywords: 486 cornea: tears/tear film/dry eye • 482 cornea: epithelium • 557 inflammation  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×