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Role of the JAK/STAT pathway in a streptozotocin-induced diabetic retinopathy mouse model

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Abstract

Purpose

The Janus tyrosine kinase and signal transducers and activators of transcription (JAK/STAT) pathway is involved in vascular endothelial growth factor (VEGF) expression, but the role of this pathway in diabetic retinopathy (DR) remains unclear. We investigated the role of the JAK/STAT pathway on DR and VEGF expression using a streptozotocin (STZ)-induced DR mouse model.

Methods

Cultured ARPE-19 cells were exposed to high-glucose conditions and treated with JAK/STAT inhibitors (JAK inhibitor I [JAKiI], tofacitinib, STAT3 inhibitor [STAT3i]) for 48 h. Reverse-transcription polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay were used to investigate p-JAK/STAT and VEGF expression. Diabetes was induced by intraperitoneal injection of STZ (50 mg/kg) in C57BL/6 mice for 5 days. DR development was evaluated every 4 weeks. JAK/STAT inhibitors were administered for 8 weeks. Immunofluorescence was used to measure the activation status of the JAK/STAT pathway and VEGF production in the retinal tissue.

Results

In ARPE-19 cells exposed to high-glucose conditions, the mRNA and secretory protein levels of VEGF, p-JAK1, p-JAK2, p-STAT3, and p-STAT5 levels were significantly increased. Treatment with JAKiI, tofacitinib, and STAT3i significantly suppressed VEGF to basal levels at both the mRNA and secretory levels in vitro. In STZ-induced mice, retinal vascular leakage, p-JAK1, p-JAK2, p-JAK3, p-STAT3, and VEGF were significantly increased after diabetes induction. Diabetes-induced retinal vascular leakage was significantly reduced by treatment with JAKiI and tofacitinib. Increased p-JAK1 and VEGF in STZ-induced mice were significantly reduced by JAKiI (p < 0.05, p < 0.001) and tofacitinib (p < 0.001, respectively).

Conclusion

JAK1 may be more involved in VEGF production and DR progression in mice than JAK2, JAK3, and STAT3.

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Data availability

Yes.

Code availability

Not applicable.

Abbreviations

DR:

Diabetic retinopathy

JAK:

Janus tyrosine kinase

PCR:

Polymerase chain reaction

RPE:

Retinal pigment epithelium

STAT:

Signal transducers and activators of transcription

STZ:

Streptozotocin

VEGF:

Vascular endothelial growth factor

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Funding

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (No.: NRF-2017R1D1A1B03033799).

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Author notes

  1. Chan-Ho Cho and Kug-Hwan Roh have contributed equally to this work.

Authors and Affiliations

  1. Department of Ophthalmology, Haeundae Paik Hospital, Inje University College of Medicine, Busan, 48108, Korea

    Chan-Ho Cho & Hyun Woong Kim

  2. Department of Microbiology and Immunology, Inje University College of Medicine, Busan, 47392, Korea

    Kug-Hwan Roh & SaeGwang Park

  3. R&D Center, NexThera Co., Ltd, Busan, 47392, Korea

    Kug-Hwan Roh, Na-Young Lim & SaeGwang Park

  4. Department of Internal Medicine, Busan Paik Hospital, Inje University College of Medicine, Busan, 47392, Korea

    Sung Jae Park

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  1. Chan-Ho Cho
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Correspondence to Hyun Woong Kim.

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All experimental procedures were approved by the Institutional Animal Care and Use Committee of Inje University (protocol no. 2017–011) and adhered to the Association for Research in Vision and Ophthalmology statement for the Use of Animals in Ophthalmic and Vision Research.

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Cho, CH., Roh, KH., Lim, NY. et al. Role of the JAK/STAT pathway in a streptozotocin-induced diabetic retinopathy mouse model. Graefes Arch Clin Exp Ophthalmol 260, 3553–3563 (2022). https://doi.org/10.1007/s00417-022-05694-7

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