Department of Ophthalmology, Jiangxi Provincial People's Hospital Affiliated to Nanchang University, Nanchang, China.
Kaohsiung J Med Sci. 2021 Feb;37(2):101-110. doi: 10.1002/kjm2.12302. Epub 2020 Oct 6.
Diabetic retinopathy (DR) is a common complication of diabetes mellitus. Inflammatory responses play crucial roles in the progress of DR. Long noncoding RNA (lncRNAs) and microRNAs (miRNAs) are key signal transduction molecules in retina, and tightly connected with DR occurrence and development. Our study aimed to explore the functions of lncRNA H19, miR-19b and silence information regulator factor related enzymes 1 (SIRT1) in the progress of DR. Retinal pigment epithelial cells (ARPE-19) were used to build high-glucose (HG) model. Quantitative real-time PCR (qPCR) was performed to detect expression of H19, miR-19b and SIRT1 at normal glucose (NG) and HG conditions. And western blotting was performed to test protein level of SIRT1. QPCR and enzyme-linked immunosorbent assay were performed to detect expression of inflammatory cytokines. Finally, the interactions among H19, miR-19b and SIRT1 were determined by dual-luciferase reporter assay. Our results showed that lncRNA H19 and SIRT1 were reduced, while miR-19b was increased in ARPE-19 cells with HG condition. MiR-19b positively regulated the expression of inflammatory cytokines, including TNF-α, IL-1β and IL-6. Inhibition of miR-19b and overexpression of H19 inhibited the expression of inflammatory cytokines, such as TNF-α, IL-1β and IL-6, while knockdown of SIRT1 reversed their effects on inflammatory cytokines. Furthermore, overexpression of miR-19b reversed the inhibitory effects of overexpression of H19 on inflammatory cytokines. Importantly, H19 targeted miR-19b to downregulate miR-19b expression. Furthermore, miR-19b bound to SIRT1 and declined SIRT1 expression. H19/miR-19b/SIRT1 axis plays a key role of HG-induced inflammatory response in ARPE-19 cells, which provides new targets for DR treatment.
糖尿病视网膜病变（DR）是糖尿病的常见并发症。炎症反应在 DR 的进展中起着至关重要的作用。长链非编码 RNA（lncRNA）和 microRNA（miRNA）是视网膜中关键的信号转导分子，与 DR 的发生和发展密切相关。我们的研究旨在探讨 lncRNA H19、miR-19b 和沉默信息调节因子相关酶 1（SIRT1）在 DR 进展中的作用。使用视网膜色素上皮细胞（ARPE-19）构建高糖（HG）模型。在正常葡萄糖（NG）和 HG 条件下，通过定量实时 PCR（qPCR）检测 H19、miR-19b 和 SIRT1 的表达。通过 Western blot 检测 SIRT1 蛋白水平。通过 qPCR 和酶联免疫吸附试验检测炎症细胞因子的表达。最后，通过双荧光素酶报告基因实验确定 H19、miR-19b 和 SIRT1 之间的相互作用。我们的结果表明，在 HG 条件下，ARPE-19 细胞中的 lncRNA H19 和 SIRT1 减少，而 miR-19b 增加。miR-19b 正向调节炎症细胞因子，包括 TNF-α、IL-1β 和 IL-6 的表达。抑制 miR-19b 和过表达 H19 抑制了 TNF-α、IL-1β 和 IL-6 等炎症细胞因子的表达，而 SIRT1 的敲低逆转了它们对炎症细胞因子的作用。此外，过表达 miR-19b 逆转了过表达 H19 对炎症细胞因子的抑制作用。重要的是，H19 靶向 miR-19b 以下调 miR-19b 的表达。此外，miR-19b 结合 SIRT1 并降低 SIRT1 的表达。H19/miR-19b/SIRT1 轴在 ARPE-19 细胞中发挥着关键作用，可作为治疗 DR 的新靶点。
