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Involvement of the JNK signaling in granular corneal dystrophy by modulating TGF-β-induced TGFBI expression and corneal fibroblast apoptosis

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Abstract

Granular corneal dystrophy (GCD) is featured by corneal deposits of transforming growth factor beta-induced gene (TGFBI) mediated by the TGF-β (transforming growth factor-β)/Smad signaling. However, the roles of c-Jun amino-terminal kinase (JNK) pathway in GCD pathogenesis remains unexplored, which was investigated in this study. JNK signaling activation and inhibition in primary corneal fibroblasts were obtained by treatments with anisomycin and SP600125, respectively. Protein abundance and phosphorylation were detected by immunoblotting. Cell viability and apoptosis were analyzed by CCK-8 and flow cytometry respectively. TGFBI deposit and autophagy progression were assessed by immunofluorescence. The results found that JNK1 expression and phosphorylation were greatly increased in corneal tissues from GCD2 patients. JNK signaling activation impaired the viability and promoted apoptosis and autophagy processes in primary corneal fibroblasts, along with Smad2/3 phosphorylation, TGFBI accumulation and Bcl-2 suppression. Autophagy related proteins, such as ATG5 (autophagy related 5), ATG12 (autophagy related 12) and LC3B (microtubule-associated protein 1 light chain 3 beta), were also increased in anisomycin or TGF-β1 treated corneal fibroblasts. However, SP600125 effectively reversed the above effect induced by TGF-β1 treatment in corneal fibroblasts, including the TGF-β-induced autophagy progression. The results suggested that JNK signaling was activated in GCD2 corneal tissues, and it mediated the TGF-β-induced TGFBI protein accumulation and apoptosis of corneal fibroblasts during GCD2 pathogenesis.

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Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (81870626), Sanming Project of Medicine in Shenzhen (SZSM201812091), and the Shenzhen Science and Technology Fund (JCYJ20160428144605809).

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Author notes

  1. Lin Ye

    Present address: Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Shenzhen University School of Medicine, No. 18, Zetian Road, Futian District, Shenzhen, 518040, China

  2. Danyao Nie and Xinhua Liu contributed equally to this work.

Authors and Affiliations

  1. Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Shenzhen University School of Medicine, No. 18, Zetian Road, Futian District, Shenzhen, 518040, China

    Danyao Nie, Xinhua Liu, Yuan Wang, Wenling He, Ming Li, Yun Peng, Jing Zhang, Liangnan Sun & Zonghui Yan

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  1. Danyao Nie
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  2. Xinhua Liu
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  3. Yuan Wang
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  4. Wenling He
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  5. Ming Li
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  10. Lin Ye
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Correspondence to Lin Ye.

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Editor: Tetsuji Okamoto

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Nie, D., Liu, X., Wang, Y. et al. Involvement of the JNK signaling in granular corneal dystrophy by modulating TGF-β-induced TGFBI expression and corneal fibroblast apoptosis. In Vitro Cell.Dev.Biol.-Animal 56, 234–242 (2020). https://doi.org/10.1007/s11626-019-00424-6

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  • DOI: https://doi.org/10.1007/s11626-019-00424-6

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