April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Ethyl Pyruvate's Effects on the TGF-B1-Induced Expression of -Smooth Muscle Actin and Fibronectin in Human Tenon's Capsule and Trabecular Meshwork Cells
Author Affiliations & Notes
  • A. B. Abbott
    Deptartment of Ophthalmology, University of Pittsburgh School of Medicine,
    UPMC Eye Center, Eye and Ear Institute, Ophthalmology and Visual Science Research Center, Pittsburgh, Pennsylvania
  • J. Chen
    Deptartment of Ophthalmology, University of Pittsburgh School of Medicine,
    UPMC Eye Center, Eye and Ear Institute, Ophthalmology and Visual Science Research Center, Pittsburgh, Pennsylvania
  • T. Stubbs
    Deptartment of Ophthalmology, University of Pittsburgh School of Medicine,
    UPMC Eye Center, Eye and Ear Institute, Ophthalmology and Visual Science Research Center, Pittsburgh, Pennsylvania
  • J. Schuman
    Dept. of Ophthalmology, U. of Pittsburgh School of Medicine, Swanson School of Engineeering.,
    UPMC Eye Center, Eye and Ear Institute, Ophthalmology and Visual Science Research Center, Pittsburgh, Pennsylvania
  • N. SundarRaj
    Deptartment of Ophthalmology, University of Pittsburgh School of Medicine,
    UPMC Eye Center, Eye and Ear Institute, Ophthalmology and Visual Science Research Center, Pittsburgh, Pennsylvania
  • Footnotes
    Commercial Relationships  A.B. Abbott, None; J. Chen, None; T. Stubbs, None; J. Schuman, Bioptigen, P; Carl Zeiss Meditec, Inc., P; Carl Zeiss Meditec, Inc., R; Heidelberg Engineering, R; Pfizer, R; N. SundarRaj, None.
  • Footnotes
    Support  P30-EY008098; Eye and Ear Foundation (Pittsburgh, PA); Research to Prevent Blindness
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 3201. doi:https://doi.org/
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      A. B. Abbott, J. Chen, T. Stubbs, J. Schuman, N. SundarRaj; Ethyl Pyruvate's Effects on the TGF-B1-Induced Expression of -Smooth Muscle Actin and Fibronectin in Human Tenon's Capsule and Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3201. doi: https://doi.org/.

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

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Abstract

Purpose: : Previously, we have shown that ethyl pyruvate (EP), a stable derivative of pyruvic acid, decreases fibroblast transformation in corneal stromal cells. The purpose of this study is to examine whether EP also exhibits anti-fibrotic effects in human Tenon's capsule and trabecular meshwork (TM) cells.

Methods: : TM and Tenon's capsule cells derived from human donors eyes were grown in culture and were treated with 15mM and 17.5mM EP. The effect of EP on cell proliferation was analyzed by immunostaining for a proliferative nuclear antigen (Ki67).The expression of alpha-smooth muscle actin (SMA) and fibronectin was evaluated by immunocytochemical and western blot analyses. Total mRNA was extracted from the cells using the Qiagen Rneasy Mini Kit. Relative abundance of mRNAs encoding fibronectin and alpha-SMA was estimated by quantitative real time RT-PCR.

Results: : EP, at 15 and 17.5 mM concentrations, inhibited TGF-beta1-induced proliferation of both TM and Tenon’s capsule cells. This was evident from lack of significant increase in cell numbers and less than 1% Ki67 positive cells after 2-4 days in culture in the presence of EP. The expression of alpha-SMA and fibronectin in both cell types was downregulated by EP as evident from decreased immunostaining and from significant loss in the intensities of the immunoreactive bands of fibronectin and alpha-SMA in the western blots of the proteins extracted from the cells. Furthermore, the levels of mRNAs encoding these proteins were lower in the EP treated cells than in the control cells, indicating that EP downregulates the transcription of fibronectin and alpha-SMA.

Conclusions: : These findings suggest that EP exhibits anti-fibrotic effects on human Tenon's capsule and trabecular meshwork cells. EP may have therapeutic potential as an agent for decreasing TGF-B1-induced fibrosis in these cells, which can occur following trabeculectomy surgery.

Keywords: trabecular meshwork • wound healing 
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