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Vitamin C alleviates hyperglycemic stress in retinal pigment epithelial cells

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Abstract

Background

Retinal pigment epithelial cells (RPECs) are a type of retinal cells that structurally and physiologically support photoreceptors. However, hyperglycemia has been shown to play a critical role in the progression of diabetic retinopathy (DR), which is one of the leading causes of vision impairment. In the diabetic eye, the high glucose environment damages RPECs via the induction of oxidative stress, leading to the release of excess reactive oxygen species (ROS) and triggering apoptosis. In this study, we aim to investigate the antioxidant mechanism of Vitamin C in reducing hyperglycemia-induced stress and whether this mechanism can preserve the function of RPECs.

Methods and results

ARPE-19 cells were treated with high glucose in the presence or absence of Vitamin C. Cell viability was measured by MTT assay. Cleaved poly ADP-ribose polymerase (PARP) was used to identify apoptosis in the cells. ROS were detected by the DCFH-DA reaction. The accumulation of sorbitol in the aldose reductase (AR) polyol pathway was determined using the sorbitol detection assay. Primary mouse RPECs were isolated from adult mice and identified by Rpe65 expression. The mitochondrial damage was measured by mitochondrial membrane depolarization. Our results showed that high glucose conditions reduce cell viability in RPECs while Vitamin C can restore cell viability, compared to the vehicle treatment. We also demonstrated that Vitamin C reduces hyperglycemia-induced ROS production and prevents cell apoptosis in RPECs in an AR-independent pathway.

Conclusions

These results suggest that Vitamin C is not only a nutritional necessity but also an adjuvant that can be combined with AR inhibitors for alleviating hyperglycemic stress in RPECs.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

AGEs:

Advanced Glycation End products

AR:

Aldose Reductase

ARPE:

Adult Retinal Pigment Epithelium

DM:

Diabetes Mellitus

DR:

Diabetic Retinopathy

FOXO3:

Forkhead box O3

PARP:

Poly (ADP-ribose) Polymerase

PKC:

Protein Kinase C

ROS:

Reactive Oxygen Species

TNF:

Tumor Necrosis Factor

VEGF:

Vascular Endothelial Growth Factor

HG:

High Glucose

LG:

Low Glucose

RPEC:

Retinal Pigment Epithelial Cell

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Acknowledgements

We thank Chandler Meadows, Hui-Chun Cheng, Soureesh Moturi, and Shining Wang for their technical assistance.

Funding

The work was supported by the NIH Core Grant P30-EY008098, the Eye and Ear Foundation of Pittsburgh, an unrestricted grant from Research to Prevent Blindness, New York, NY, the Shaffer grant from Glaucoma Research Foundation, and the Biomedical Masters Program at University of Pittsburgh. VYL was supported by Hillman Academy.

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Author notes

  1. Hamid Alahmari and Chia-Chun Liu contributed equally to this work.

Authors and Affiliations

  1. Department of Ophthalmology, Louis J. Fox Center for Vision Restoration, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15219, USA

    Hamid Alahmari, Chia-Chun Liu, Elizabeth Rubin, Venice Y. Lin, Paul Rodriguez & Kun-Che Chang

  2. Department of Neurobiology, Center of Neuroscience, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213, USA

    Kun-Che Chang

  3. Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan

    Kun-Che Chang

  4. North Allegheny Senior High School, Wexford, PA, 15090, USA

    Venice Y. Lin

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  1. Hamid Alahmari
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Contributions

HA, CCL, ER, VYL conducted experiments and analyzed data; HA, PR and KCC wrote the manuscript; KCC supervised the whole project.

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Correspondence to Kun-Che Chang.

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Alahmari, H., Liu, CC., Rubin, E. et al. Vitamin C alleviates hyperglycemic stress in retinal pigment epithelial cells. Mol Biol Rep 51, 637 (2024). https://doi.org/10.1007/s11033-024-09595-2

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