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Moesin Expression in Fibrosis in the Mouse Cornea After Sterile Mechanical Trauma or Infection

Zhu, Hong-Y. MD, PhD*; Ng, Jennifer BSc*; Salleh, Shuhaida M. BSc*; Aung, Thet T. MBBS, MSc*; Htoon, Myint H. BDS, PhD*; Beuerman, Roger W. PhD*,†

Cornea:
doi: 10.1097/ICO.0000000000000179
Basic Investigation
Abstract

Purpose: The aim of this study was to compare the expression patterns of 3 important biochemical characteristics of fibrosis–moesin, transforming growth factor (TGF)-β1, and α-smooth muscle actin (SMA) in the mouse cornea with fibrosis induced by common etiologies—sterile mechanical injury and infection.

Methods: Corneas of 8-week-old C57BL6 mice were either wounded after an anterior keratectomy or were infected by Pseudomonas aeruginosa, and the animals were killed on days 2 and 7, and on weeks 2 and 4 after the procedure. Western blot and immunofluorescence were used to analyze the expression of moesin and phospho-moesin, and the presence of myofibroblasts identified by the expression of α-SMA in the corneal stroma. The expression of TGF-β1 was analyzed by immunofluorescence.

Results: By immunofluorescent analysis, TGF-β1, α-SMA, and phospho-moesin were not detected in the normal corneal stroma. However, after either treatment, TGF-β1 expression increased, along with phospho-moesin in the wounded corneal stroma until day 7, and decreased after week 2. No expression of TGF-β1 and phospho-moesin was found at postoperative week 4. Moesin expression increased until week 2. Myofibroblasts positive for α-SMA were detected on day 2 until week 4 and peaked at week 2. Western blot analysis confirmed the immunofluorescent data for moesin, phospho-moesin, and α-SMA.

Conclusions: The similar expression pattern of moesin, phospho-moesin, TGF-β1, and α-SMA in the mouse cornea with fibrosis caused by sterile mechanical injury or infection indicated a role for moesin signaling in corneal fibrosis. Interference with the action of moesin may be a potential approach for intervention strategies to avert fibrosis after infection or mechanical injury.

Author Information

*Singapore Eye Research Institute, Singapore, Singapore; and

SRP Neuroscience and Behavioral Disorders, Duke-NUS, Singapore, Singapore.

Reprints: Roger W. Beuerman, PhD, Singapore Eye Research Institute, 11 Third Hospital Ave, Singapore 168751, Singapore (e-mail: rwbeuerman@gmail.com).

The authors have no conflicts of interest to disclose.

Supported by a Singapore National Medical Research Council Center Grant, Singapore Eye Research Institute Pilot grant R934/43/2012.

Received January 17, 2014

Accepted May 06, 2014

© 2014 by Lippincott Williams & Wilkins.