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Metformin protects retinal pigment epithelium cells against H2O2-induced oxidative stress and inflammation via the Nrf2 signaling cascade

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Abstract

Purpose

Dysfunctions of retinal pigment epithelium (RPE) attributed to oxidative stress and inflammation are implicated with age-related macular degeneration (AMD). A debate on the curative role of metformin in AMD has been raised, though several recent clinical studies support the lower odds by using metformin. This study aimed to determine whether metformin could exert cytoprotection against RPE oxidative damages and the potential mechanisms.

Methods

A cellular AMD model was established by treating ARPE-19 cells with hydrogen peroxide (H2O2) for 24 h. The reactive oxygen species (ROS) generation, expression of antioxidant enzymes, and levels of pro-inflammatory cytokines were monitored under administrations with H2O2 with/without metformin. The expression and DNA-binding activity of transcription factor erythroid-related factor 2 (Nrf2) were determined by western blot, immunofluorescence, and electrophoretic mobility shift assay. Knockout of Nrf2 was conducted by CRISPR/Cas9 gene deletion system.

Results

Metformin pretreatment significantly improved the H2O2-induced low viability of ARPE-19 cells, reduced ROS production, and increased contents of antioxidative molecules. Concurrently, metformin also suppressed levels of pro-inflammatory cytokines caused by H2O2. The metformin-augmented nuclear translocation and DNA-binding activity of Nrf2 were further verified by the increased expression of its downstream targets. Genetic deletion of Nrf2 blocked the cytoprotective role of metformin.

Conclusion

Metformin possesses antioxidative and anti-inflammatory properties in ARPE-19 cells by activating the Nrf2 signaling. It supports the potential use for the control and prevention of AMD.

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Funding

This study was funded by the National Natural Science Foundation of China (82171209) and the Guangzhou Science Technology and Innovation Commission (202102010019).

National Natural Science Foundation of China,82171209,Yuehong Zhang,Guangzhou Science,Technology and Innovation Commission,202102010019,Yuehong Zhang.

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Authors and Affiliations

  1. State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, China

    Qiting Feng

  2. Department of Anesthesia and Pain Medicine, The Sixth Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China

    Xiangcai Ruan

  3. Sanshui Huaxia Eye Hospital, Huaxia Eye Hospital Group, Foshan, China

    Min Lu

  4. Department of Ophthalmology, Guangzhou First People’s Hospital, School of Medicine, South China University of Technology, Guangzhou, 510080, China

    Shimiao Bu & Yuehong Zhang

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  1. Qiting Feng
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  2. Xiangcai Ruan
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  3. Min Lu
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  4. Shimiao Bu
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  5. Yuehong Zhang
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Contributions

Qiting Feng and Yuehong Zhang designed the research; Qiting Feng, Xiangcai Ruan, and Min Lu performed experiments and collected data; Qiting Feng, Xiangcai Ruan, and Shimiao Bu analyzed data; Qiting Feng, Xiangcai Ruan, Min Lu, Shimiao Bu, and Yuehong Zhang wrote the paper. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Yuehong Zhang.

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Feng, Q., Ruan, X., Lu, M. et al. Metformin protects retinal pigment epithelium cells against H2O2-induced oxidative stress and inflammation via the Nrf2 signaling cascade. Graefes Arch Clin Exp Ophthalmol 262, 1519–1530 (2024). https://doi.org/10.1007/s00417-023-06321-9

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