June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Antifibrotic effect of rapamycin, an inhibitor of mTOR pathway, on the TGF-β2 induced proliferation of human tenon fibroblasts
Author Affiliations & Notes
  • Shaodan Zhang
    Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, Liaoning, China
  • Lin Du
    Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, Liaoning, China
  • Di Wu
    Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, Liaoning, China
  • Chi Liu
    Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, Liaoning, China
  • Hailin Wang
    Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, Liaoning, China
  • Footnotes
    Commercial Relationships   Shaodan Zhang, None; Lin Du, None; Di Wu, None; Chi Liu, None; Hailin Wang, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4948. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Shaodan Zhang, Lin Du, Di Wu, Chi Liu, Hailin Wang; Antifibrotic effect of rapamycin, an inhibitor of mTOR pathway, on the TGF-β2 induced proliferation of human tenon fibroblasts. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4948.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Bleb failure by fibrotic scarring remains a major problem of glaucoma filtration surgery. Dysfunctional PI3K/mTOR signaling has recently been recognized as a driver of aberrant proliferative responses in kidney, pulmonary and cardiac fibrosis. In this study, we investigated the anti-fibrotic effect of rapamycin, an mTOR inhibitor, on the TGF-b2 induced proliferation of human tenon fibroblasts (HTFs).

Methods : Human tenon tissue was obtained from adult strabismus patients during surgery. Primary cell culture for the HTFs was performed. After 24 hour incubation with 2ng/mL TGF-β2, the cells were treated with rapamycin 1nM for 48 hours. Cell viability and proliferation were measured with MTT assay. Cell migration was assessed by scratch wound assay. Flow cytometry of AnnexinV/propidium iodide (PI) staining was performed to measure the cell apoptosis. Expression of proliferative gene MKi67, pro-apoptotic gene caspase 3, and autophagy related gene LC3B were analyzed by real-time PCR.

Results : TGF-β2 significantly promoted the proliferation (138% of controls, p<0.01) and migration of HTFs, as well as the mRNA expression of MKi67. Rapamycin effectively attenuate the TGF-β2 induced cell proliferation (80.8% of TGF-β2gourp), and migration. The up-regulated MKi67 expression in TGF-β2 treated HTFs was also distinctively inhibited by rapamycin. Cell apoptosis, as assessed by both flow cytometry and caspase 3 mRNA expression, was not observed in all groups. As an autophagy inducer, rapamycin significantly up-regulated the mRNA expression of LC3B, an important autophagy marker, in the HTF cells with and without TGF-β2.

Conclusions : Rapamycin could effectively prevent the TGF-β2 induced proliferation and migration of HTF cells without affecting the cell survival. The involvement of mTOR and the underlying mechanism of autophagy need further investigation. Rapamycin may become a potential anti-fibrotic agent that may facilitate the therapeutic effect of glaucoma filtering surgery.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

 

 

×
×

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.

×