Department of Ophthalmology, The First affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China.
IUBMB Life. 2019 Oct;71(10):1611-1618. doi: 10.1002/iub.2102. Epub 2019 Jul 18.
High-glucose-induced retinal tissue impairment is the major pathological phenotype of diabetic retinopathy. In an in vitro diabetic apoptosis cell model, we evaluated the function of long noncoding RNA, insulin growth factor 2 antisense (IGF2-AS) in high-glucose-injured human retinal pigment epithelial cells. A human retinal pigment epithelial cell line, ARPE-19 was incubated with high-glucose in vitro to induce apoptosis. SiRNA-mediated IGF2-AS downregulation was conducted in ARPE-19 cells to evaluate its effect on high-glucose induced apoptosis, assessed by a TUNEL assay. qRT-PCR and western blot assays were applied to examine the functional effect of IGF2-AS on IGF2/AKT/Casp-9 expressions in glucose-injured ARPE-19 cells. ART was further knocked down, specifically in IGF2-AS-downregualted ARPE-19 cells, to investigate its functional involvement in IGF2-AS-inhibition-mediated apoptotic protection in glucose-injured ARPE-19 cells. High-glucose induced apoptosis in ARPE-19 cells, and upregulated IGF-2AS in a dose-dependent manner. SiRNA-mediated IGF2-AS downregulation ameliorated apoptosis, upregulated IGF2/AKT and decreased Casp-9, in high-glucose-treated ARPE-19 cells. AKT knockdown was shown to dramatically reverse the preventive effect of IGF2-AS-downregulation on high-glucose-induced apoptosis in ARPE-19 cells. Moreover, it was demonstrated that AKT knockdown directly upregulated Casp-9 in IGF2-AS-downregulated and high-glucose-treated ARPE-19 cells. We demonstrated that inhibiting IGF2-AS, possibly also through activation of AKT signaling pathway, has a protective function in high-glucose-induced apoptosis in human retinal pigment epithelial cells in diabetic retinopathy.
高糖诱导的视网膜组织损伤是糖尿病视网膜病变的主要病理表型。在体外糖尿病细胞凋亡模型中，我们评估了长非编码 RNA 胰岛素样生长因子 2 反义（IGF2-AS）在高糖损伤的人视网膜色素上皮细胞中的功能。体外将人视网膜色素上皮细胞系 ARPE-19 与高糖孵育以诱导细胞凋亡。在 ARPE-19 细胞中用 siRNA 介导 IGF2-AS 下调，通过 TUNEL 测定评估其对高糖诱导的细胞凋亡的影响。应用 qRT-PCR 和 Western blot 检测 IGF2-AS 对葡萄糖损伤的 ARPE-19 细胞中 IGF2/AKT/Casp-9 表达的功能影响。进一步敲低 ART，特别是在 IGF2-AS 下调的 ARPE-19 细胞中，以研究其在 IGF2-AS 抑制介导的葡萄糖损伤的 ARPE-19 细胞凋亡保护中的功能参与。高糖诱导 ARPE-19 细胞凋亡，并呈剂量依赖性地上调 IGF-2AS。siRNA 介导的 IGF2-AS 下调可改善高糖处理的 ARPE-19 细胞中的细胞凋亡，上调 IGF2/AKT，并降低 Casp-9。AKT 敲低可显著逆转 IGF2-AS 下调对 ARPE-19 细胞中高糖诱导的细胞凋亡的预防作用。此外，还证明 AKT 敲低可直接上调 IGF2-AS 下调和高糖处理的 ARPE-19 细胞中的 Casp-9。我们证明抑制 IGF2-AS，可能通过激活 AKT 信号通路，在糖尿病视网膜病变中具有高糖诱导的人视网膜色素上皮细胞凋亡的保护功能。
