摘要
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.
摘要译文
糖尿病性视网膜病（DR）是糖尿病并发症（DM）的并发症，可能会导致严重的视觉损失。视网膜色素上皮（RPE）在视网膜生理学中起着至关重要的作用，但容易受到氧化损伤的影响。我们研究了硒（SE）对视网膜色素上皮（ARPE-19）和原发性人视网膜微血管内皮（ACBRI 181）细胞对高葡萄糖（HG）诱导的氧化应激和凋亡级联反应的保护作用。为了实现这一目标，我们利用了不同浓度的D-葡萄糖（从5到80 mm）来诱导HG模型。通过确定CA2+过载，线粒体膜去极化，CASPASE-3/-9激活，细胞内活性氧，ROS），脂质过氧（ROS），脂质过化（LP），脂质过氧（LP），脂质性氧气（LP），脂质诱导的ARPE-19和ACBRI 181细胞中HG诱导的氧化应激以及凋亡级联反应。谷胱甘肽（GSH），谷胱甘肽过氧化物酶（GSH-PX），血管内皮生长因子（VEGF）和凋亡水平。使用MTT进行了细胞活力测定，以确定要使用的SE的最佳浓度。使用板读取器完成了MTT，ROS，VEGF水平和caspase -3和-9激活的定量。为了定量评估LP和GSH水平，分光光度计和凋亡，线粒体膜去极化利用GSH-PX活性，并通过光谱荧光仪评估了Ca2+从细胞内存储中释放。我们的研究表明，通过调节线粒体膜电位，ROS水平和细胞内Ca2+释放，导致细胞损伤导致细胞损伤。与SE一起孵育导致HG诱导的ROS产生显着降低，以及凋亡的降低和Caspase -3和-9的激活。此外，SE孵育导致VEGF和LP的水平降低，同时增加GSH和GSH-PX水平。这项研究的发现强烈表明，SE对ARPE-19和ACBRI 181细胞具有保护作用，以针对HG诱导的氧化应激和凋亡。该保护机制通过细胞内CA2+信号通路部分介导。
Handan Bardak;Abdülhadi Cihangir Uğuz;Yavuz Bardak;Javier Rocha-Pimienta;Jonathan Delgado-Adámez;Javier Espino. Selenium Protects ARPE-19 and ACBRI 181 Cells against High Glucose-Induced Oxidative Stress[J]. Molecules Online, 2023,28(16): 5961