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Lobeglitazone attenuates fibrosis in corneal fibroblasts by interrupting TGF-beta-mediated Smad signaling

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Abstract

Purpose

Transforming growth factor beta 1 (TGF-β1) is an important cytokine released after ocular surface injury to promote wound healing. However, its persistence at the injury site triggers a fibrotic response that leads to corneal scarring and opacity. Thiazolidinediones (TZDs) are synthetic peroxisome proliferator-activated receptor gamma (PPAR-γ) ligands used to regulate glucose and lipid metabolism in the management of type 2 diabetes. Studies have also showed TZDs have antifibrotic effect. In this study, we investigated the antifibrotic effect of the TZD lobeglitazone on TGF-β1-induced fibrosis in corneal fibroblasts.

Methods

Human primary corneal fibroblasts were cultivated and treated with TGF-β1 (5 ng/mL) to induce fibrosis, with or without pre-treatments with different concentrations of lobeglitazone. Myofibroblast differentiation and extracellular matrix (ECM) protein expression was evaluated by western blotting, immunofluorescence, real-time PCR, and collagen gel contraction assay. The effect of lobeglitazone on TGF-β1-induced reactive oxygen species (ROS) generation was evaluated by DCFDA–cellular ROS detection assay kit. Signaling proteins were evaluated by western blotting to determine the mechanism underlying the antifibrotic effect.

Results

Our results showed lobeglitazone attenuated TGF-β1-induced ECM synthesis and myofibroblast differentiation of corneal fibroblasts. This antifibrotic effect appeared to be independent of PPAR signaling and rather due to the inhibition of the TGF-β1-induced Smad signaling. Lobeglitazone also blocked TGF-β1-induced ROS generation and nicotinamide adenine dinucleotide phosphate oxidase (Nox) 4 transcription.

Conclusion

These findings indicate that lobeglitazone may be a promising therapeutic agent for corneal scarring.

Key messages

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Funding

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology (2017R1D1A3B03036549 and 2020R1I1A3073515).

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Authors and Affiliations

  1. Department of Global Medical Science, Wonju College of Medicine, Yonsei University, Wonju, Ilsan-ro, Gangwon-do, 26426, Republic of Korea

    Selikem Nuwormegbe

  2. Department of Ophthalmology, Wonju College of Medicine, Yonsei University, Wonju, Ilsan-ro, Gangwon-do, 26426, Republic of Korea

    Na-Young Park & Sun Woong Kim

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  1. Selikem Nuwormegbe
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  2. Na-Young Park
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Contributions

Conceptualization, Sun Woong Kim; Funding acquisition, Sun Woong Kim; Supervision, Sun Woong Kim; Investigation, Selikem Nuwormegbe; Methodology, Selikem Nuwormegbe and Sun Woong Kim; Formal analysis, Selikem Nuwormegbe and Na-Young Park; Writing original draft, Selikem Nuwormegbe; Writing—review and editing, Na-Young Park and Sun Woong Kim

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Correspondence to Sun Woong Kim.

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Nuwormegbe, S., Park, NY. & Kim, S.W. Lobeglitazone attenuates fibrosis in corneal fibroblasts by interrupting TGF-beta-mediated Smad signaling. Graefes Arch Clin Exp Ophthalmol 260, 149–162 (2022). https://doi.org/10.1007/s00417-021-05370-2

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  • DOI: https://doi.org/10.1007/s00417-021-05370-2

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