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CREG Protects Retinal Ganglion Cells loss and Retinal Function Impairment Against ischemia-reperfusion Injury in mice via Akt Signaling Pathway

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Abstract

Purpose

The irreversible death of retinal ganglion cells (RGCs) plays an important role in the pathogenesis of glaucoma. Cellular repressor of E1A-stimulated genes (CREG), a secreted glycoprotein involved in cellular proliferation and differentiation, has been shown to protect against myocardial and renal ischemia‐reperfusion damage. However, the role of CREG in retinal ischemia-reperfusion injury (RIRI) remains unknown. In this study, we aimed to explore the effect of CREG on RGCs apoptosis after RIRI.

Methods

We used male C57BL/6J mice to establish the RIRI model. Recombinant CREG was injected at 1 day before RIRI. The expression and distribution of CREG were examined by immunofluorescence staining and western blotting. RGCs survival was assessed by immunofluorescence staining of flat-mounted retinas. Retinal apoptosis was measured by the staining of TdT-mediated dUTP nick-end labeling and cleaved caspase-3. Electroretinogram (ERG) analysis and optomotor response were conducted to evaluate retinal function and visual acuity. The expressions of Akt, phospho-Akt (p-Akt), Bax, and Bcl-2 were analyzed by western blotting to determine the signaling pathways of CREG.

Results

We found that CREG expression was decreased after RIRI, and intravitreal injection of CREG attenuated RGCs loss and retinal apoptosis. Besides, the amplitudes of a-wave, b-wave, and photopic negative response (PhNR) in ERG, as well as visual function, were significantly restored after treatment with CERG. Furthermore, intravitreal injection of CREG upregulated p-Akt and Bcl-2 expression and downregulated Bax expression.

Conclusion

Our results demonstrated that CREG protected RGCs from RIRI and alleviated retinal apoptosis by activating Akt signaling. In addition, CREG also improved retinal function and visual acuity.

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Data availability

All the data and results used in this study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Jiayi Yang, Xiao Zhang, Ying Li, and Lina Pan for their constructive suggestions.

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Author notes

  1. Siyu Zeng, Lei Du contributed equally to this work.

Authors and Affiliations

  1. Eye Center, Renmin Hospital of Wuhan University, 238 Jiefang Road, Wuhan, 430060, Hubei Province, People’s Republic of China

    Siyu Zeng, Lei Du, Guojing Lu & Yiqiao Xing

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Contributions

S.Z and L.D designed the study and conducted the experiments. G.L and Y.X reviewed and revised the manuscript. All authors have approved the final article and agree with submission of the manuscript.

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Correspondence to Yiqiao Xing.

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This study was performed in line with the principles of the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Approval was granted by the Ethics Committee of the Renmin Hospital of Wuhan University.

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Zeng, S., Du, L., Lu, G. et al. CREG Protects Retinal Ganglion Cells loss and Retinal Function Impairment Against ischemia-reperfusion Injury in mice via Akt Signaling Pathway. Mol Neurobiol 60, 6018–6028 (2023). https://doi.org/10.1007/s12035-023-03466-w

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