Upregulated VEGF and Robo4 correlate with the reduction of miR-15a in the development of diabetic retinopathy
Upregulated VEGF and Robo4 correlate with the reduction of miR-15a in the development of diabetic retinopathy
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DOI：
10.1007/s12020-019-01921-0
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发表时间：
2019-07-01
期刊：
影响因子：
3.7
通讯作者：
Su, Guanfang
中科院分区：
文献类型：
--
作者：
Gong, Qiaoyun;Li, Fuqiang;Su, Guanfang
目的血管内皮生长因子(VEGF)在糖尿病视网膜病变(DR)中起重要作用。环状4(Robo4)在血管生成和血管生成中的作用一直存在争议，然而，这两个因子之间的相互依赖关系在DR中还没有研究。本研究确定了增殖性糖尿病视网膜病变(PDR)患者纤维血管膜(FVM)中VEGF和Robo4的共存。方法采用免疫荧光法检测糖尿病大鼠和ARPE-19细胞在高血糖状态下血管内皮生长因子(VEGF)和Robo4的表达。用Western blotting和RT-qPCR方法检测高糖条件下糖尿病视网膜和ARPE-19细胞中血管内皮生长因子和Robo4的表达。通过眼内注射microRNA agomir来增加糖尿病视网膜miR-15a的表达，下调VEGF和Robo4的水平。在高血糖条件下培养的糖尿病视网膜和ARPE-19组织培养细胞中，血管内皮生长因子水平的升高是一致的。高血糖作用72小时后，ARPE-19组织培养细胞中Robo4表达减少，而糖尿病大鼠视网膜中Robo4表达增加。在DR进展过程中，几个miRNAs的表达存在差异。此外，miR-15a琼脂糖凝胶可抑制糖尿病视网膜组织中高水平的血管内皮生长因子和Robo4的表达。在糖尿病视网膜病变早期，血管内皮生长因子表达上调，参与了糖尿病视网膜病变的发生发展，而在糖尿病视网膜病变晚期，血管内皮生长因子和Robo4共同作用，加速了糖尿病视网膜病变的发展。但是，miR-15a可以下调VEGF和Robo4的表达，从而改善DR的发展。
PurposeVascular endothelial growth factor (VEGF) plays implicated roles in diabetic retinopathy (DR). The role of roundabout 4 (Robo 4) in angiogenesis and vasculogenesis is controversial; however, the interdependent relationship between these two factors has not been studied in DR. This study determined the colocalization of VEGF and Robo4 in fibrovascular membranes (FVM) from patients with proliferative diabetic retinopathy (PDR). MicroRNA (miRNA)-mediated modulation of VEGF and Robo4 was explored in diabetic rats and ARPE-19 tissue culture cells under hyperglycemia.MethodsVEGF and Robo4 co-expression in the FVM was analyzed using immunofluorescence. VEGF and Robo4 levels were determined in diabetic retinas and ARPE-19 tissue culture cells under high glucose using western blotting and RT-qPCR. MicroRNA agomir was intraocularly injected to increase miR-15a expression and downregulate VEGF and Robo4 levels in diabetic retinas.ResultsVEGF and Robo4 colocalization in FVM vessels was observed. Increased VEGF levels were consistent in diabetic retinas and ARPE-19 tissue culture cells cultured under hyperglycemia. Robo4 decreased in ARPE-19 tissue culture cells exposed to hyperglycemia for 72h, whereas it increased in diabetic rat retinas. Several miRNAs were differentially expressed during DR progression. Furthermore, miR-15a agomir injection inhibited high levels of VEGF and Robo4 in diabetic retinas.ConclusionsVEGF and Robo4 were co-expressed in FVMs from PDR patients. In the early stages of DR, VEGF was upregulated and contributed to DR development, whereas, in the late stage of DR, VEGF and Robo4 worked together to aggravate DR progression. However, miR-15a could downregulate VEGF and Robo4 to ameliorate DR development.