Abstract
Chronic hyperglycemia involves persistent high-glucose exposure and correlates with retinal degeneration. It causes various diseases, including diabetic retinopathy (DR), a major cause of adult vision loss. Most in vitro studies have investigated the damaging short-term effects of high glucose exposure on retinal pigment epithelial (RPE) cells. DR is also a severe complication of diabetes. In this study, we established a model with prolonged high-glucose exposure (15 and 75 mM exogenous glucose for two months) to mimic RPE tissue pathophysiology in patients with hyperglycemia. Prolonged high-glucose exposure attenuated glucose uptake and clonogenicity in ARPE-19 cells. It also significantly increased reactive oxygen species levels and decreased antioxidant protein (superoxide dismutase 2) levels in RPE cells, possibly causing oxidative stress and DNA damage and impairing proliferation. Western blotting showed that autophagic stress, endoplasmic reticulum stress, and genotoxic stress were induced by prolonged high-glucose exposure in RPE cells. Despite a moderate apoptotic cell population detected using the Annexin V-staining assay, the increases in the senescence-associated proteins p53 and p21 and SA-β-gal-positive cells suggest that prolonged high-glucose exposure dominantly sensitized RPE cells to premature senescence. Comprehensive next-generation sequencing suggested that upregulation of oxidative stress and DNA damage-associated pathways contributed to stress-induced premature senescence of ARPE-19 cells. Our findings elucidate the pathophysiology of hyperglycemia-associated retinal diseases and should benefit the future development of preventive drugs.
Graphical Abstract
Prolonged high-glucose exposure downregulates glucose uptake and oxidative stress by increasing reactive oxygen species (ROS) production through regulation of superoxide dismutase 2 (SOD2) expression. Autophagic stress, ER stress, and DNA damage stress (genotoxic stress) are also induced by prolonged high-glucose exposure in RPE cells. Consequently, multiple stresses induce the upregulation of the senescence-associated proteins p53 and p21. Although both apoptosis and premature senescence contribute to high glucose exposure-induced anti-proliferation of RPE cells, the present work shows that premature senescence rather than apoptosis is the dominant cause of RPE degeneration, eventually leading to the pathogenesis of DR.
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The data that support the findings of this study are available on request from the corresponding author, Prof. Sheu, upon reasonable request.
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This work was supported by: the Ministry of Science and Technology, Taiwan (Grant numbers MOST 109-2314-B-037-024, MOST 109-2313-B-037-001, MOST 109-2314-B-037-069-MY3, NSTC 111-2314-B-037-067 and NSTC 112-2314-B-037-029); NSYSU-KMU (joint grants, Grant number NSYSUKMU110-P024); the Kaohsiung Medical University Hospital, Taiwan (Grants KMUH109-9R51 and KMUH110-0M47 and KMUH111-1R46) and the Kaohsiung Medical University Research Center, Taiwan (Grant number KMU-TC112A04).
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C-CC and S-JS designed the experiments and wrote the paper. K-CC conceptualization. Y-HL, C-XH, and Y-DB performed the experiments and collected the data. Y-HL and Y-DB performed the bioinformatic analysis. C-CC and S-JS supply the funding acquisition. K-CC provided the cell line. K-CC, C-YL, C-YW, and H-MW provided reagents and translational suggestions. C-CC and S-JS wrote the draft. C-CC and S-JS checked and edited the manuscript. All authors approved the manuscript.
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Chiu, CC., Cheng, KC., Lin, YH. et al. Prolonged Exposure to High Glucose Induces Premature Senescence Through Oxidative Stress and Autophagy in Retinal Pigment Epithelial Cells. Arch. Immunol. Ther. Exp. 71, 21 (2023). https://doi.org/10.1007/s00005-023-00686-9
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DOI: https://doi.org/10.1007/s00005-023-00686-9