Cited by 87Glaucoma and retinal disordersTGF-β signaling in diseasesIntraocular Surgery and Lenses

Suppression of TGF-β pathway by pirfenidone decreases extracellular matrix deposition in ocular fibroblasts in vitro

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Stahnke, Thomas, Kowtharapu, Bhavani S., Stachs, Oliver, Schmitz, Klaus-Peter, Wurm, Johannes, Wree, Andreas, Guthoff, Rudolf Friedrich, Hovakimyan, Marina·2017·PLOS ONE

Abstract

In glaucoma surgery, fibrotic processes occur, leading to impairment of liquid outflow. Activated fibroblasts are responsible for postoperative scarring. The transforming growth factor-β (TGF-β) pathway plays a key role in fibroblast function, differentiation and proliferation. The aim of this study was the characterization of the fibrotic potential of two subtypes of primary human ocular fibroblasts and the attempt to inhibit fibrotic processes specifically, without impairing cell viability. For fibrosis inhibition we focused on the small molecule pirfenidone, which has been shown to prevent pulmonary fibrosis by the decrease of the expression of TGF-β1, TGF-β2 and TGF-β3 cytokines. For in vitro examinations, isolated human primary fibroblasts from Tenon capsule and human intraconal orbital fat tissues were used. These fibroblast subpopulations were analyzed in terms of the expression of matrix components responsible for postoperative scarring. We concentrated on the expression of collagen I, III, VI and fibronectin. Additionally, we analyzed the expression of α-smooth muscle actin, which serves as a marker for fibrosis and indicates transformation of fibroblasts into myofibroblasts. Gene expression was analyzed by rtPCR and synthesized proteins were examined by immunofluorescence and Western blot methods. Proliferation of fibroblasts under different culture conditions was assessed using BrdU assay. TGF-β1 induced a significant increase of cell proliferation in both cell types. Also the expression of some fibrotic markers was elevated. In contrast, pirfenidone decreased cell proliferation and matrix synthesis in both fibroblast subpopulations. Pirfenidone slightly attenuated TGF-β1 induced expression of fibronectin and α-smooth muscle actin in fibroblast cultures, without impairing cell viability. To summarize, manipulation of the TGF-β signaling pathway by pirfenidone represents a specific antifibrotic approach with no toxic side effects in two human orbital fibroblast subtypes. We presume that pirfenidone is a promising candidate for the treatment of fibrosis following glaucoma surgery.

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Crossref · 10.1371/journal.pone.0172592

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  • Pirfenidone reduces TGF-β signaling and inflammatory cytokine production in different murine experimental systems.

    “Consistent with this, the small molecule pirfenidone (PFD), has been shown to reduce TGF-β signaling and inflammatory cytokine production in different murine experimental systems (135–137)”

    From Targeting macrophages in liver fibrosis (2026) · cites this paper as [136]

    The Abstract and Discussion report antifibrotic effects of pirfenidone involving TGF-β in human ocular fibroblast cultures. They do not report reduced inflammatory cytokine production or experiments in murine systems; those parts of the attributed claim are unsupported by this paper. (LLM, conf=0.88)

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  1. [4]Quigley HA. Glaucoma. The Lancet 377 2011:(9774): 1367– 1377. 21453963

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Authors
Stahnke, T., Kowtharapu, B. S., Stachs, O., Schmitz, K.-P., Wurm, J., Wree, A., Guthoff, R. F., & Hovakimyan, M.
Affiliations
Universitätsmedizin Rostock (DE)
Corresponding author
Stahnke, Thomas
Venue
PLOS ONE
Published
2017-02-23
Volume
12
Issue
2
Pages
e0172592
ISSN
1932-6203 (ISSN-L)
OpenAlex
W2591068902
Cited by
87
References
45
Citations by year
2026: 2 · 2025: 4 · 2024: 8 · 2023: 10 · 2022: 6 · 2021: 12 · 2020: 16 · 2019: 12 · 2018: 12 · 2017: 5
FWCI
12.00
Type
journal article
Language
en
Open access
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • PUB – Publications at Bielefeld University (Bielefeld University) (submittedVersion)
Funders
Bundesministerium für Bildung und Forschung (03IS2081)
Topics
  • Health Sciences › Medicine › Ophthalmology › Glaucoma and retinal disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › TGF-β signaling in diseases
  • Health Sciences › Medicine › Ophthalmology › Intraocular Surgery and Lenses
Keywords
pirfenidone, TGF-beta signaling, ocular fibroblasts, postoperative scarring, glaucoma surgery, fibrosis, extracellular matrix deposition, collagen, fibronectin, alpha-smooth muscle actin, myofibroblast differentiation, TGF-β1, antifibrotic therapy, orbital fat
MeSH terms
Actins, Anti-Inflammatory Agents, Non-Steroidal, Cells, Cultured, Extracellular Matrix, Fibroblasts, Fibronectins, Fibrosis, Gene Expression Regulation, Humans, Pyridones, Signal Transduction, Transforming Growth Factor beta, Cell Proliferation

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