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Inhibition of TLR4 alleviates the inflammation and apoptosis of retinal ganglion cells in high glucose

  • Basic Science
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Abstract

Purpose

To investigate the expression profiles of Toll-like receptor 4 (TLR4), the effect of TLR4 on inflammation, and apoptosis of retinal ganglion cells (RGCs) cultured in high glucose and the underlying mechanism.

Methods

A high-glucose model was established in RGCs isolated from Sprague-Dawley (SD) rats (2–3 days old) and identified with Brn3a. Primary cultured RGCs were divided into control (0 mM), HG1 (10 mM glucose), HG2 (20 mM glucose), HG3 (30 mM glucose), HG (20 mM glucose) + TAK-242 (1.0 μM), and HG (20 mM glucose) + vehicle (1% DMSO) groups. The expression levels of TLR4, its downstream signalling molecules, and pro-inflammatory cytokines were measured by real-time PCR, Western blot or ELISA at 24 h and 48 h. The apoptosis rate of RGCs was measured by flow cytometry.

Results

The mRNA and protein expression levels of TLR4 were increased in high-glucose groups (10 mM, 20 mM, 30 mM). Consistent with these findings, four TLR4 downstream signalling molecules (MyD88, NF-κB, TRAF6, NLRP3) and pro-inflammatory cytokines (IL-1β, IL-18) were upregulated in the three high-glucose groups. Apoptosis of RGCs was clearly increased in the high-glucose group. The administration of TAK-242, an antagonist of TLR4, inhibited inflammation and apoptosis of RGCs in the high-glucose group.

Conclusion

Our results demonstrated that TLR4 plays a critical role in the inflammation and apoptosis of RGCs induced by high glucose. TLR4 might become a novel potential pharmacological target for preventing the progression of DR.

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Authors and Affiliations

  1. Eye Center, Renmin Hospital of Wuhan University, 238 Jiefang road, Wuchang district, Wuhan, Hubei, 430060, China

    Lili Hu, Hongxia Yang, Ming Ai & Shuanghong Jiang

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  1. Lili Hu
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  2. Hongxia Yang
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  3. Ming Ai
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Contributions

Hongxia Yang and Lili Hu conceived and designed the project. Lili Hu performed most of the experiments and wrote the manuscript. Lili Hu and Shuanghong Jiang analysed the data. Ming Ai and Hongxia Yang reviewed and revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Ming Ai.

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All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest or non-financial interest in the subject matter or materials discussed in this manuscript.

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The funding organizations had no role in the design or conduct of this research.

Animal experiments

All procedures performed in the studies were in accordance with the ethical standards of the institution and national research committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

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Hu, L., Yang, H., Ai, M. et al. Inhibition of TLR4 alleviates the inflammation and apoptosis of retinal ganglion cells in high glucose. Graefes Arch Clin Exp Ophthalmol 255, 2199–2210 (2017). https://doi.org/10.1007/s00417-017-3772-0

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