The hsa_circ_0004805/hsa_miR-149-5p/TGFB2 axis plays critical roles in the pathophysiology of diabetic retinopathy in vitro and in vivo
The hsa_circ_0004805/hsa_miR-149-5p/TGFB2 axis plays critical roles in the pathophysiology of diabetic retinopathy in vitro and in vivo
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DOI：
10.1016/j.mce.2023.112042
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发表时间：
2023-08-14
影响因子：
4.1
通讯作者：
Xin，Yu
中科院分区：
文献类型：
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作者：
Shao，Jun;Gu，Wendong;Xin，Yu
本研究的目的是探讨最近发现的hsa_circ_0004805/hsa_miR-149-5p/转化生长因子β 2（TGFB2）轴在糖尿病视网膜病变（DR）进展中的作用机制。定量逆转录聚合酶链反应（qRT-PCR）分析表明，hsa_circ_0004805在DR患者房水样品中高表达，而hsa_miR-149-5p呈相反趋势。同时，双荧光素酶报告基因检测结果表明，hsa_miR-149-5p直接与hsa_circ_0004805和tgfb2相互作用。通过多种检测（Cell Counting Kit-8、edu_labeling、Transwell、流式细胞术、伤口愈合、管形成检测），我们发现在高糖环境下培养的人视网膜微血管上皮细胞（hRECs）中，hsa_circ_0004805过表达显著下调hsa_miR-149-5p水平，促进DNA合成、增殖、迁移和管形成。相反，hsa_miR-149-5p模拟物通过降低其下游靶标ttgfb2的表达以及磷酸化SMAD2的水平，抑制hRECs中的DNA合成、增殖、迁移和管形成；然而，这些影响被hsa_circ_0004805的过表达逆转。在链脲佐菌素诱导的DR大鼠Sprague-Dawley模型中，视网膜血管渗漏、毛细血管脱细胞、周细胞丢失、纤维化和胶质细胞形成明显，这可以通过玻璃体显微注射大鼠miR-149-5p模拟物（rno-miR-149-5p agomir）来逆转。综上所述，我们的研究结果表明，在高血糖状态下，hsa_circ_0004805/hsa_miR-149-5p/ tgfb2轴在与DR发展相关的视网膜病理生理中起着关键作用，并有可能成为治疗该疾病的治疗靶点。
The aim of this study was to investigate the mechanism underlying the role of a recently identified hsa_circ_0004805/hsa_miR-149-5p/transforming growth factor beta 2(TGFB2)axis in the progression of diabetic retinopathy (DR). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis suggested that hsa_circ_0004805 was highly expressed in aqueous humor samples of patients with DR, whereas hsa_miR-149-5p showed the opposite trend. Meanwhile, the results of a dual-luciferase reporter assay indicated that hsa_miR-149-5p directly interacted with both hsa_circ_0004805 andTGFB2. Using a variety of assays (Cell Counting Kit-8, EdU-labeling, Transwell, flow cytometric, wound healing, tube formation assays), we found that the overexpression of hsa_circ_0004805 significantly downregulated the level of hsa_miR-149-5p and promoted DNA synthesis, proliferation, migration, and tube formation in human retinal microvascular epithelial cells (hRECs) cultivated in a high-glucose environment. In contrast, hsa_miR-149-5p mimics inhibited DNA synthesis, proliferation, migration, and tube formation in hRECs by reducing the expression of its downstream targetTGFB2as well as the levels of phosphorylated SMAD2; however, these effects were reversed by the overexpression of hsa_circ_0004805. In a streptozotocin-induced Sprague-Dawley rat model of DR, retinal vascular leakage, capillary decellularization, loss of pericytes, fibrosis, and gliosis were evident, which could be reversed by vitreous microinjection of rat miR-149-5p mimics (rno-miR-149-5p agomir). Combined, our findings indicated that, under hyperglycemia, the hsa_circ_0004805/hsa_miR-149-5p/TGFB2axis plays a critical role in the retinal pathophysiology associated with the development of DR, and has potential as a therapeutic target in the treatment of this condition.