Department of Ophthalmology, First Affiliated Hospital of Zhengzhou University, Henan Province Eye Hospital, Zhengzhou, 450052, China.
Department of Ophthalmology, First Affiliated Hospital of Zhengzhou University, Henan Province Eye Hospital, Zhengzhou, 450052, China.
Biomed Pharmacother. 2020 May;125:109959. doi: 10.1016/j.biopha.2020.109959. Epub 2020 Feb 25.
Patients with chronic hyperglycemia are at high risk of developing diabetic retinopathy. In this study, we investigated the functional role of long-noncoding RNA (lncRNA) X-inactive specific transcript (XIST) in anin vitro model of diabetic hyperglycemia in human retinal pigment epithelial ARPE-19 cells.
ARPE-19 cells were cultured in normal glucose (NG) and high-glucose (HG) conditions to mimic hyperglycemia-associated cell apoptosis, migration and XIST expression. XIST was overexpressed in ARPE-19 cells to examine its functions in HG-induced cell apoptosis and migration. The downstream competing target of XIST, human mature microRNA-21-5p (hsa-miR-21-5p) was assessed by dual-luciferase assay and qRT-PCR. Hsa-miR-21-5p was upregulated in XIST-overexpressed ARPE-19 cells to further assess the functional correlation between XIST and hsa-miR-21-5p in hyperglycemia-associated cell apoptosis and migration.
HG insult increased apoptosis, reduced migration and downregulated XIST in ARPE-19 cells. XIST overexpression significantly protected HG insult in ARPE-19 cells, by reducing apoptosis and restoring migration capability. XIST directly bound and inhibited hsa-miR-21-5p expression in HG-insulted ARPE-19 cells. Furthermore, hsa-miR-21-5p upregulation reversed the protective effects of XIST in HG-insulted ARPE-19 cells.
XIST, likely through competitive binding of hsa-miR-21-5p, provides protection against hyperglycemia-associated injury in human retinal pigment epithelial cells.
患有慢性高血糖的患者患糖尿病性视网膜病变的风险很高。在这项研究中，我们研究了长非编码 RNA（lncRNA）X 失活特异性转录物（XIST）在人视网膜色素上皮 ARPE-19 细胞体外高血糖模型中的功能作用。
将 ARPE-19 细胞在正常葡萄糖（NG）和高葡萄糖（HG）条件下培养，以模拟与高血糖相关的细胞凋亡、迁移和 XIST 表达。在 ARPE-19 细胞中转染 XIST 以观察其在 HG 诱导的细胞凋亡和迁移中的作用。通过双荧光素酶报告基因和 qRT-PCR 评估 XIST 的下游竞争靶标，人成熟 microRNA-21-5p（hsa-miR-21-5p）。在 XIST 过表达的 ARPE-19 细胞中上调 hsa-miR-21-5p，以进一步评估 XIST 和 hsa-miR-21-5p 在高血糖相关细胞凋亡和迁移中的功能相关性。
HG 刺激增加了 ARPE-19 细胞的凋亡，减少了迁移，并下调了 XIST。XIST 的过表达可显著减轻 HG 对 ARPE-19 细胞的损伤，减少凋亡并恢复迁移能力。XIST 直接结合并抑制 HG 刺激的 ARPE-19 细胞中 hsa-miR-21-5p 的表达。此外，hsa-miR-21-5p 的上调逆转了 XIST 在 HG 刺激的 ARPE-19 细胞中的保护作用。
XIST 可能通过与 hsa-miR-21-5p 的竞争性结合，为人类视网膜色素上皮细胞提供了对抗高血糖相关损伤的保护。
