Selenium Protects ARPE-19 and ACBRI 181 Cells against High Glucose-Induced Oxidative Stress

Handan Bardak; Abdülhadi Cihangir Uğuz; Yavuz Bardak; Javier Rocha-Pimienta; Jonathan Delgado-Adámez; Javier Espino

doi:10.3390/molecules28165961

Selenium Protects ARPE-19 and ACBRI 181 Cells against High Glucose-Induced Oxidative Stress

Molecules. 2023 Aug 9;28(16):5961. doi: 10.3390/molecules28165961.

Authors

Handan Bardak¹, Abdülhadi Cihangir Uğuz², Yavuz Bardak³, Javier Rocha-Pimienta⁴, Jonathan Delgado-Adámez⁴, Javier Espino⁵

Affiliations

¹ Department of Ophthalmology, Asya Hospital, 34100 Istanbul, Turkey.
² Department of Biophysics, Faculty of Medicine, Karamanoğlu Mehmetbey University, 70100 Karaman, Turkey.
³ Worldeye Hospital, 34337 Istanbul, Turkey.
⁴ Technological Agri-Food Institute (CICYTEX-INTAEX), Junta of Extremadura, Avda. Adolfo Suárez s/n, 06007 Badajoz, Spain.
⁵ Department of Physiology, Faculty of Science, University of Extremadura, Avda. de Elvas, s/n, 06006 Badajoz, Spain.

Abstract

Diabetic retinopathy (DR), a complication of diabetes mellitus (DM), can cause severe visual loss. The retinal pigment epithelium (RPE) plays a crucial role in retinal physiology but is vulnerable to oxidative damage. We investigated the protective effects of selenium (Se) on retinal pigment epithelium (ARPE-19) and primary human retinal microvascular endothelial (ACBRI 181) cells against high glucose (HG)-induced oxidative stress and apoptotic cascade. To achieve this objective, we utilized varying concentrations of D-glucose (ranging from 5 to 80 mM) to induce the HG model. HG-induced oxidative stress in ARPE-19 and ACBRI 181 cells and the apoptotic cascade were evaluated by determining Ca²⁺ overload, mitochondrial membrane depolarization, caspase-3/-9 activation, intracellular reactive oxygen species (ROS), lipid peroxidation (LP), glutathione (GSH), glutathione peroxidase (GSH-Px), vascular endothelial growth factor (VEGF) and apoptosis levels. A cell viability assay utilizing MTT was conducted to ascertain the optimal concentration of Se to be employed. The quantification of MTT, ROS, VEGF levels, and caspase-3 and -9 activation was accomplished using a plate reader. To quantitatively assess LP and GSH levels, GSH-Px activities were utilized by spectrophotometer and apoptosis, mitochondrial membrane depolarization, and the release of Ca²⁺ from intracellular stores were evaluated by spectrofluorometer. Our investigation revealed a significant augmentation in oxidative stress induced by HG, leading to cellular damage through modulation of mitochondrial membrane potential, ROS levels, and intracellular Ca²⁺ release. Incubation with Se resulted in a notable reduction in ROS production induced by HG, as well as a reduction in apoptosis and the activation of caspase-3 and -9. Additionally, Se incubation led to decreased levels of VEGF and LP while concurrently increasing levels of GSH and GSH-Px. The findings from this study strongly suggest that Se exerts a protective effect on ARPE-19 and ACBRI 181 cells against HG-induced oxidative stress and apoptosis. This protective mechanism is partially mediated through the intracellular Ca²⁺ signaling pathway.

Keywords: ARPE-19 cells; high glucose; oxidative stress; selenium.

MeSH terms

Caspase 3
Glucose / toxicity
Humans
Oxidative Stress
Reactive Oxygen Species
Selenium* / pharmacology
Vascular Endothelial Growth Factor A

Substances

Selenium
Vascular Endothelial Growth Factor A
Caspase 3
Reactive Oxygen Species
Glucose

Grants and funding

This research was financially supported by the first author herself.